FIELD OF THE INVENTION

The present invention relates to heterocyclic compounds useful as anti-infective agents. The present invention further relates to a method of treating an infection by administering such a compound. The present invention further relates to pharmaceutical compositions comprising such compounds. BACKGROUND ART

Antimicrobial resistance is an increasingly serious threat to global public health. New resistance mechanisms emerge and spread globally, threatening the effective prevention and treatment of a range of infections caused by bacteria, parasites and fungi. A number of examples can be provided to illustrate the threat posed. In 2013 there was approximately half a million new cases of multi-drug resistant tuberculosis. Resistance to artemisinin-based combination therapies, which are the best available treatment for falciparum malaria, has been detected in the Greater Mekong subregion. Highly resistant bacteria such as MRSA cause a high percentage of hospital-acquired infections. Patients with such drug-resistant infections have an increased risk of inferior clinical outcomes and death as compared to patients infected with non-resistant bacteria. Ten countries have reported cases where gonorrhoea was untreatable due to resistance to the treatmetns of last resort (3 generation cephalosporins). Thus, gonorrhoea may soon become untreatable. There is thus an increased and urgent need for new anti-infective agents for use in therapy.

SUMMARY OF THE INVENTION

The object of the invention is thus to provide compounds useful for the treatment or prevention of infection. A further object is to provide a method of treating an infection, such as a bacterial, fungal or parasitic infection.

These objects are achieved by compounds as disclosed by the appended claims. The compounds have the Formula I:

or a pharmaceutically acceptable salt thereof wherein

A is selected from S and O; each of X ¹ , X ² , X ³ , X ⁴ and X ⁵ is independently selected from C and N;

R ¹ is selected from the group consisting of

-H, -Ci-6 alkyl,

-Ci-6 alkyl-amino wherein the amino group is optionally substituted with one or twoCi_ ₆ acyl or Ci- ₆ alkyl groups, and

-Ci-6 alkyl-heterocyclyl wherein the heterocyclyl group is a 5- or 6- membered aliphatic or aromatic heterocycle, optionally benzo-fused, and is optionally substituted with one or more R ⁶ groups;

R ² is selected from the group consisting of-H, -CF ₃ , -N0 ₂ , -N(R ⁵ ) ₂ , , -NHR ⁵ ,—

N(R ⁵ )C(0)R ⁵ , and -N(R ⁵ )C(S)N(R ⁵ ) ₂ ; or

1 2

R and R together with the atoms to which they are bound form a 5- or 6-membered fused heterocyclic ring substituted with one or more R ⁵ groups; R ³ is selected from -CF ₃ , -CN, -CI, -Ci- ₆ alkyl, -C _3-6 cycloalkyl, -C(0)NH ₂ , -C(0)NH-Ci- ₆ alkyl, -NH-heterocyclyl, -phenyl, and -heterocyclyl, wherein the heterocyclyl group is a 5- or 6- membered aliphatic or aromatic optionally benzo-fused heterocycle, and wherein R ³ is optionally substituted with one of more R ⁶ groups; each of R ⁴ and R ⁸ is selected from H, -CN, -halo, -CF ₃ , -Ci- ₆ alkoxy, -C0 ₂ -Ci- ₆ alkyl, -N0 ₂ , - Ci-6 alkyl-NH ₂ , -heterocyclyl, and -CONH _m [(CH ₂ )„NH ₂ ] _2-m , wherein the heterocyclyl group is a 5- or 6- membered aliphatic or aromatic optionally benzo-fused heterocycle; each instance of R ⁵ is independently selected from the group consisting of

-H, -Ci-6 alkyl optionally substituted with one or more R ⁶ groups, -C ₂ -6 alkenyl optionally substituted with one or more R ⁶ groups,

-Co-3 alkyl-C3-6 cycloalkyl-Co-3 alkyl optionally substituted with one or more R ⁶ groups, -phenyl optionally substituted with one or more R ⁶ groups, -C=C-Ph optionally substituted with one or more R ⁶ groups, and

-Co-3 alkyl-heterocyclyl-Co-3 alkyl optionally substituted with one or more R ⁶ groups, wherein the heterocyclyl group is a 5- ,6- or 7-membered aliphatic or aromatic optionally benzo-fused heterocycle; each instance of R ⁶ is independently selected from the group consisting of -halo, -CN, -Ci- ₆ alkyl, -OH, -Ci- ₆ alkoxy, -Ci _-6 alkyl-NH ₂ , -NH _m [(CH ₂ ) _n NH ₂ ] _2-m , -NH ₂ , -NH-Ci _-6 alkyl, and - N-Ci _-6 dialkyl; n and m are integers, wherein each instance of n is independently chosen from 2 or 3, and each instance of m is independently chosen from 0 or 1 ;

2 1

with the proviso that if R is -H, then R is not -H or -C1-6 alkyl. Compounds, or salts therefore, as defined by Formula I can be used in the treatment or prevention of infection, especially bacterial infection.

Without wishing to be bound by theory, it is thought that the compounds disclosed above achieve their antimicrobial effect at least in part by inhibition of RNase P. RNase P is a ribonucleoprotein complex present in all living cells. It catalyses the removal of 5'leader sequences from tRNA precursors and similar molecules. In bacteria, RNase P consists of one RNA subunit and a small basic protein, and it has been shown that the catalytic activity is associated with its RNA subunit. RNase P is potentially a good drug target since RNase P is indispensable for bacterial viability and the architecture of RNase P differs between bacteria and eukaryote. For example, the important P-15 loop in bacteria is a good target for antibacterial drug design.

The compounds of Formula I may belong to a subset of compounds having a Formula II:

(II)

or a pharmaceutically acceptable salt thereof.

The compounds of Formula I or II may have a structure wherein:

A is selected from S and O;

X ¹ is selected from C and N;

R ¹ is selected from the group consisting of

-H,

Ci-3 alkyl,

-Ci-3 alkyl-amino wherein the amino group is optionally substituted with one or two acetyl or Ci-3 alkyl groups, and -Ci-3 alkyl-heterocyclyl wherein the heterocyclyl group is selected from imidazolyl, piperazinyl and thiomorpholinyl and is optionally substituted with one or more R ⁶ groups;

R ² is selected from the group consisting of-H, -CF ₃ , -N0 ₂ , -N(R ⁵ ) ₂ , , -NHR ⁵ ,—

N(R ⁵ )C(0)R ⁵ , and -N(R ⁵ )C(S)N(R ⁵ ) ₂ ; or

R ¹ and R ² together with the atoms to which they are bound form a 5- or 6-membered fused heterocyclic ring substituted with one or more R ⁵ groups;

R ³ is selected from -CF ₃ , -CN, -CI, -Ci- ₃ alkyl, -C _3-6 cycloalkyl, -C(0)NH ₂ , -C(0)NH-Ci- ₃ alkyl, -NH-piperazinyl, -phenyl, -pyridinyl, -indolyl, -benzimidazolyl, -benzothiazolyl, and - benzopyrazolyl, wherein R ³ is optionally substituted with one of more R ⁶ groups; each of R ⁴ and R ⁸ is selected from H, -CN, -CI, -F, -CF ₃ , -Ci- ₃ alkoxy, -C0 ₂ Me, -N0 ₂ , -Ci- ₃ alkyl-NH ₂ , -piperazinyl, -indolyl, and -CONH _m [(CH ₂ ) _n NH ₂ ] _2-m ; each instance of R ⁵ is independently selected from the group consisting of

-H, -Ci-3 alkyl optionally substituted with one or more R ⁶ groups, -C ₂ -3 alkenyl optionally substituted with one or more R ⁶ groups,

-Co-3 alkyl-C ₃ -6 cycloalkyl-Co-3 alkyl optionally substituted with one or more R ⁶ groups, -phenyl optionally substituted with one or more R ⁶ groups, -C=C-Ph optionally substituted with one or more R ⁶ groups, and

-Co-3 alkyl-heterocyclyl-Co-3 alkyl optionally substituted with one or more R ⁶ groups, wherein the heterocyclyl group is selected from azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl and indolyl; each instance of R ⁶ is independently selected from the group consisting of -F, -CI, -CN, -C1-3 alkyl, -OH, -C1-3 alkoxy, -C1-3 alkyl-NH ₂ , -NH _m [(CH ₂ ) _n NH ₂ ] _2-m , -NH ₂ , -NHMe, and -NMe ₂ ; n and m are integers, wherein each instance of n is independently chosen from 2 or 3, and each instance of m is independently chosen from 0 or 1 ;

2 1

with the proviso that if R is -H, then R is not -H or -C1-3 alkyl. The compounds of Formula I or II may have a structure wherein R ² is selected from the group consisting of-H, -CF ₃ , -N(R ⁵ ) ₂ , -NHR ⁵ , -N(R ⁵ )C(0)R ⁵ , and -N(R ⁵ )C(S)N(R ⁵ ) ₂ ; with the proviso that if R ² is -H, then R ¹ is not -H or -C ₁ -3 alkyl and R ⁸ is not H.

The compounds of Formula I or II may have a structure wherein R ⁸ is not H. Thus, the compounds of the invention may have substituents at the R ¹ , R ³ and R ⁸ positions, or

2 3 8

alternatively at the R , R and R . It has been found that thus substituted compounds are especially active in the treatment or prevention of infection.

The compounds of Formula I or II may have a structure wherein X ¹ , X ² , X ³ , X ⁴ and X ⁵ are C.

The compounds of Formula I or II may have a structure wherein R ¹ is H. The compounds of Formula I or II may have a structure wherein R ² is selected from the group consisting of-NH ₂ and -NHR ⁵ .

2 5

The compounds of Formula I or II may have a structure wherein R is -NHC(0)R .

The compounds of Formula I or II may have a structure wherein R ² is H.

The compounds of Formula I or II may have a structure wherein R ⁴ is H. The compounds of Formula I or II may have a structure wherein A is S.

The compounds of Formula I or II may have a structure wherein R ³ is selected from the group consisting of-CF ₃ and -indolyl.

According to another aspect of the present invention, the objects of the invention are achieved by a compound according to Formula I or II as disclosed above, for use in a method of treatment of the human or animal body by therapy. The therapy may be treatment or prevention of an infection. The infection may be a bacterial, fungal, or parasitic infection. The infection may be a bacterial infection caused or complicated by bacteria of a genus selected from Staphylococcus, Enterococcus, Streptococcus, Pseudomonas, Legionella, Klebsiella, Haemophilus, Neisseria, Listeria,Escherichia d Mycobacterium. The bacterial infection may be caused or complicated by a bacterial species selected from the group: S. aureus, E. faecalis, E. faecium, S. pneumoniae, E. coli, K. pneumoniae, H. influenza, A. baumannii, P. aeruginosa, P. aeruginosa, N. gonorrhoeae. The bacterial infection may be caused or complicated by a bacterial species selected from the group: M.fortuitum, M. phlei, M. tuberculosis.

According to a further aspect of the present invention, the objects of the invention are achieved by a method of treating an infection which comprises administering to a patient in need thereof a therapeutically effective amount of a compound as disclosed above. The infection may be a bacterial, fungal, or parasitic infection. The infection may be a bacterial infection caused or complicated by bacteria of a genus selected from Staphylococcus, Enterococcus, Streptococcus, Pseudomonas, Legionella, Klebsiella, Haemophilus, Neisseria, Listeria,Escherichia d

Mycobacterium. The bacterial infection may be caused or complicated by a bacterial species selected from the group: S. aureus, E. faecalis, E. faecium, S. pneumoniae, E. coli, K.

pneumoniae, H. influenza, A. baumannii, P. aeruginosa, P. aeruginosa, N. gonorrhoeae. The bacterial infection may be caused or complicated by a bacterial species selected from the group: M. fortuitum, M. phlei, M. tuberculosis.

According to yet another aspect of the present invention, the object of the invention is achieved by use of a compound as disclosed above, or a salt thereof, in inhibition of bacterial RNase P activity.

According to yet a further aspect of the present invention, the object of the invention is achieved by use of a compound as disclosed above, or a salt thereof, as a bactericide.

According to still a further aspect of the present invention, the object of the invention is achieved by a pharmaceutical composition comprising a compound as disclosed above, or a

pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable excipient, adjuvant, diluent and/or carrier.

Further aspects, objects and advantages are defined in the detailed description below with reference to the appended drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

For the understanding of the present invention and further objects and advantages of it, the detailed description set out below can be read together with the accompanying drawings. Fig. 1 shows general synthetic scheme I for the synthesis of selected compounds according to the present invention.

Fig. 2 shows general synthetic scheme II for the synthesis of selected compounds according to the present invention.

Fig. 3 shows general synthetic scheme ΠΙ for the synthesis of selected compounds according to the present invention.

Fig. 4 shows general synthetic scheme IV for the synthesis of selected compounds according to the present invention.

Fig. 5 shows general synthetic scheme V for the synthesis of selected compounds according to the present invention.

Fig. 6 shows general synthetic scheme VI for the synthesis of selected compounds according to the present invention.

Fig. 7 shows general synthetic scheme VII for the synthesis of selected compounds according to the present invention.

Fig. 8 shows general synthetic scheme VIII for the synthesis of selected compounds according to the present invention.

Fig. 9 shows general synthetic scheme ΓΧ for the synthesis of selected compounds according to the present invention.

Fig. 10 shows general synthetic scheme X for the synthesis of selected compounds according to the present invention.

Fig. 11 shows general synthetic scheme XT for the synthesis of selected compounds according to the present invention.

Fig. 12 shows general synthetic scheme XII for the synthesis of selected compounds according to the present invention.

Fig. 13 shows general synthetic scheme XIII for the synthesis of selected compounds according to the present invention.

Fig. 14 shows general synthetic scheme XIV for the synthesis of selected compounds according to the present invention.

Fig. 15 shows general synthetic scheme XV for the synthesis of selected compounds according to the present invention.

Fig. 16 shows general synthetic scheme XVI for the synthesis of selected compounds according to the present invention. shows general synthetic scheme XXIII for the synthesis of selected compounds accordingto the present invention.

shows general synthetic scheme XXIV for the synthesis of selected compounds according to the present invention.

shows general synthetic scheme XXV for the synthesis of sleeted compounds according to the present invention

shows general synthetic scheme XXVI for the synthesis of sleeted compounds according to the present invention

shows general synthetic scheme XXVII for the synthesis of sleeted compounds according to the present invention

shows general synthetic scheme XXVTQ for the synthesis of sleeted compounds according to the present invention

shows general synthetic scheme XXVTX for the synthesis of sleeted compounds according to the present invention

shows general synthetic scheme XXX for the synthesis of sleeted compounds according to the present invention

shows general synthetic scheme XXXI for the synthesis of sleeted compounds according to the present invention

shows general synthetic scheme XXXII for the synthesis of sleeted compounds according to the present invention

shows general synthetic scheme XXXIII for the synthesis of sleeted compounds according to the present invention

DETAILED DESCRD7TION General Synthetic methods

All reactions were carried out under dry nitrogen and or argon atmosphere unless otherwise specified. Unless otherwise stated, all the raw starting materials, solvents, and reagents were purchased from commercial sources (e.g., AVRA Chemicals, Apollo Scientific Limited, Bepharma Ltd., Combi-Blocks Inc., Sigma Aldrich Chemicals Pvt. Ltd., Ultra Labs, Toronto Research Chemicals Inc., Chemical House, RFCL Limited, Spectro Chem Pvt. Ltd., Leonid Chemicals, Loba Chemie, Changzhou Yangyuan, NeoSynth., Rankem, etc.) and used as such without further purification. Alternatively, reagents may be synthesized by procedures known in the literature.

The following abbreviations are used and have the indicated definitions: MHz is megahertz (frequency), m is multiplet, t is triplet, d is doublet, s is singlet, br is broad, CDCI ₃ is deutero chloroform, calcd is calculated, min is minutes, h is hours, g is grams, mmol is millimoles, mL is milliliters, N is normality (concentration), M is molarity (concentration), μΜ is micromolar, ee is enantiomeric excess, de is diastereomeric excess, °C is degree centigrade, HPLC is High Performance Liquid Chromatography, LC-MS is Liquid Chromatography-Mass Spectroscopy, NMR is Nuclear Magnetic Resonance, TLC is Thin Layer Chromatography, THF is tetrahydrofuran, MeOH is methanol, DCM is dichloromethane, DEA is diethylamine, DMA is dimethylacetamide, DMF is N,N-dimethyl formamide, DMSO is dimethyl sulfoxide, EtOH is ethyl alcohol, EtOAc is ethyl acetate, RT is room temperature, HC1 is hydrogen chloride or hydrochloric acid, TFA is trifluoroacetic acid, EtMgBr is ethyl magnesium bromide, w-BuLi is n- butyl lithium, NaHCC is sodium bicarbonate, Na ₂ CC>3 is sodium carbonate, Na ₂ S0 ₄ is sodium sulfate, DCC is N,N-dicyclohexylcarbodiimide, DIPA is diisopropylamine, LDA is lithium diisopropylamine, HOBt is N-hydroxy-benzotriazole, NCS is N-chlorosuccinimide, and TBAB is tetrabutyl ammonium bromide.

Biotage Isolera® One and CombiFlash®(Teledyne Isco) Automated Flash Purification System were used for the purification of crude products using the eluent combination mentioned in the respective procedures. Flash Chromatography was performed using silica gel (60-100, 100-200 and 230-400 mesh) from ChemLabs, with nitrogen and/or compressed air. Preparative thin-layer chromatography was carried out using silica gel (GF 1500 μΜ 20 x 20 cm and GF 2000 μΜ 20 x 20 cm prep-scored plates from Analtech, Inc. Delaware, USA). Thin-layer chromatography was carried out using pre-coated silica gel sheets (Merck 60 F ₂ 5 ₄ ). Visual detection was performed with ultraviolet light, /7-anisaldehyde stain, ninhydrin stain, dinitrophenyl hydrazine stain, potassium permanganate stain, or iodine. Reactions at lower temperature were performed by using cold baths, e.g., H ₂ 0/ice at 0°C, and acetone/dry ice at - 78°C. Melting points were determined by using a Lablndia MR-VIS visual melting range apparatus. 1H NMR spectra were recorded at 400 MHz with a Varian V400 spectrometer, Bruker 400 (unless otherwise noted) at ambient temperature, using tetramethylsilane as internal reference. The chemical shift values are quoted in δ (parts per million). Mass spectra of all the intermediates and final compounds were recorded using Acquity® UPLC-SQD (Waters) & Agilent 1290 Infinity® with 6150 SQD machines. HPLC spectra were recorded using Agilent 1290 Infinity® UHPLC and Alliance (Waters) systems. LCMS spectra were recorded using Agilent 1200® LCMS/Agilent 1290® UHPLC-SQD with diode array detector (DAD) detection LC-MS instruments using Kinetex C18 (50 mm x 2.1mm x 2.7mic)and/orX-terra MS C18 (50mm x 2.1mm x 3.0micron) columns. The purity of each of the final compounds was detected using Waters® PDA with SQD or Aglient® DAD with 6150 SQD instrument.

The compounds according to Formulas I & II are prepared using conventional organic synthetic methods. A suitable synthetic route is depicted below in the following general reaction Schemes. The skilled artisan will appreciate that if a substituent described herein is not compatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions. The protecting group may be removed at a suitable point in the reaction sequence to provide a desired intermediate or target compound. Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Organic Synthesis(4th ed.), John Wiley & Sons, NY (2006). In some instances, a substituent may be specifically selected to be reactive under the reaction conditions used. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful as an intermediate compound or is a desired substituent in a target compound.

Description of Synthesis

Figure 1 shows general synthetic scheme I for the synthesis of selected 3, N-10-disubstituted phenothiazenes. Nucleophilic substitution of 2-amino thiophenol (Ia)with aryl halides (lb) resultedin the corresponding thioethers(Ic). N-formylation followed by Smiles rearrangement of thioethers (Ic) yielded 3 -substituted phenothiazenes (Ie). N-alkylation of Ie with distinct alkyl halides using NaH as baseyielded the corresponding N-10-alkylated phenothiazenes (If&Ih). The mono haloalkylated phenothiazenes (If) were reacted with nucleophiles (amines & alcohols) followed by salt preparations using HC1 to give the corresponding salts (Ih). In case of ester analogues of Ih, ester hydrolysis followed by amide formation yieldedthe title compounds (Ii). Figure 2 shows general synthetic scheme II for the synthesis of selectedl, 3, N-10-trisubstituted phenothiazenes. Nucleophilic substitution of 2-amino thiophenol (Ila) with aryl halides (Ilb)followed by insituSmiles rearrangement yielded 1 ,3-disubstituted phenothiazenes (lie), which are reduce by Pd/C to yield the corresponding 1 -amino subtituted phenothiazenes (lid). The phenothiazenes (lid) were selectively alkylated at N-10 position with distinct alkyl halides using NaH as base yilded correspondingN-10-alkylated phenothiazines (He). The N-10- alkylated phenothiazenes (He) were reacted with amines yielded Ilf. Amide formation of amines (Ilf) by reacting with acid chlorides or acids yielded corresponding amide analogues (Ilg). Deprotection of Ilgusing HC1 resulted the title compounds ΙΠι. Ilf reaction with CSC1 ₂ followed by reaction with NH ₃ resulted in title compounds Ilh.

A detailed synthetic description of a compound synthesised by the method of Scheme II follows. Compound 87: l-(10-(3-(Dimethylamino)propyl)-3-(trifluoromethyl)-10H-phen othiazin-l- yl)thiourea

Step 1: l-Nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-chloro-l ,3-dinitro-5-(trifluoromethyl)benzene (20 g, 74.0 mmol) in ethanol (300 mL) were added 2-aminobenzenethiol (8.0 mL, 74.0 mmol), sodium hydroxide (8.8 g, 222 mmol) and the reaction mixture was heated at 85 °C for 12 h. The reaction mixture was cooled to room temperature, concentrated, and the residue was washed with EtOH followed by H ₂ 0 to give the title compound as a brown solid (16.0 g, 70%): ¾ NMR (DMSO-d _6, 400 MHz) δ 6.97 (m, 1H), 7.02 (m, 1H), 7.09 (m, 2H), 7.65 (s, 1H), 8.01 (s, 1H), 9.84 (s, 1H); MS (ESI) m/z 311 (M-H) ⁺ .

Step 2: 3-(Trifluoromethyl)-10H-phenothiazin-

To a stirred solution of l-nitro-3-(trifluoromethyl)-10H-phenothiazine(5.0 g, 16.2 mmol) in MeOH (30 mL) was added 10% Pd/C (50% wet, 0.4 g) and the reaction mixture was stirred at room temperature for 5 h. The reaction mixture was filtered through celite and filtrate was concentrated to give the title compound as a light pink solid (6.0 g, 66%): 1H NMR (DMSO-d6, 400 MHz) δ 5.44 (s, 2H), 6.49 (s, 1H), 6.73 (s, 1H), 6.81 (t, J = 7.2 Hz, 1H), 6.92 (d, J = 8 Hz, 1H), 6.92 (d, J= 7.6 Hz, 1H), 7.01 (t, J= 7.6 Hz, 1H), 7.81 (s, 1H); MS (ESI) m/z 281 (M-H) ⁺ .

Step 3 : 10-(3-Iodopropyl)-3-(trifluoromethyl)- 1 OH-phenothiazin- 1-amine

To a stirred solution of 3-(trifluoromethyl)-10H-phenothiazin-l -amine (2.5 g, 8.86 mmol) in DMF (15mL) at 0 °C under nitrogen atmosphere was added sodium hydride (0.5 g, 12.41 mmol) and stirred for 30 min. followed by the addition of 1,3-diiodopropane (1.2 mL, 10.63 mmol). The reaction mixture was stirred at room temperature for 30 min, quenched with saturated ammonium chloride solution, extracted with ethyl acetate and concentrated. The crude productwas purified by column chromatography over silica gel using ethyl acetate/hexane (1 :19) mixture as eluent to give the title compound (2.7 g, crude) as a blue solid: 1H NMR

(DMSO-d _6, 400 MHz) δ 3.11 (t, J = 6.0 Hz, 2H), 4.81 (t, J = 6.0 Hz, 2H), 6.87 (m, 2H), 6.97 (d, J = 7.2 Hz, 1H), 7.06 (t, J = 7.6 Hz, 1H), 7.17 (s, 1H), 7.29 (s, 1H), 7.96 (s, 1H), 9.66 (s, 1H); MS (ESI) «¾/ 451(M+H) ⁺ . Step 4: 10-(3-(Dimethylamino)propyl)-3-(trifluoromethyl)-10H-phenoth iazin-l-amine

To a stirred solution of 10-(3-iodopropyl)-3-(trifluoromethyl)-10H-phenothiazin-l -amine (1.7 g, 3.77 mmol) in DMF (10 mL) were added potassium iodide (1.6 g, 11.33 mmol), dimethylamine in THF (0.25 mL, 5.66 mmol) and stirred at room temperature for 12 h. The reaction mixture was concentrated and the residue was purified by column chromatography over silica gel using methanol/dichloromethane (1 : 10) mixture as eluent to give the title compound as angreen solid (0.26 g, 31%): 1H NMR (DMSO-d _6, 400 MHz) δ 1.61 (m, 2H), 2.07 (s, 6H), 2.32 (m, 2H), 3.79 (m, 2H), 5.49 (s, 2H), 6.65 (s, 1H), 6.89 (s, 1H), 7.01 (t, J= 6.8 Hz, 1H), 7.21 (m, 3H); MS (ESI) m/z 368 (M+H) ⁺ .

Step 5: 3-(l-Isothiocyanato-3-(trifluoromethyl)-10H-phenothiazin-10- yl)-/V^V- dimethylpropan-l-amine

To a stirred solution of 10-(3-(dimethylamino)propyl)-3-(trifluoromethyl)-10H-phenoth iazin-l- amine (0.1 g, 0.272 mmol) in chloroform (5 mL) and water (5 mL) was added sodium bicarbonate solution (0.025 g, 0.68 mmol) under cooling condition, the reaction mixture was stirred at same temperature for 1 h,thiophosgene (0.037 g, 0.326 mmol) was added,the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with water, extracted with ethyl acetate, and concentrated. Thecrude productwas purified by column chromatography over silica gel using methanol/dichloromethane (1 :9) mixture as eluent to give the title compound (0.06 g, crude): MS (ESI) m/z 410.1 (M+H) . The compound was used in next step without further purification.

Step 6: l-(10-(3-(Dimethylamino)propyl)-3-(trifluoromethyl)-10H-phen othiazin-l- yl)thiourea

To a stirred solution of 3-(l -isothiocyanato-3-(trifluoromethyl)-10H-phenothiazin-10-yl)- N _r N- dimethylpropan-l-amine(0.025 g, 0.61 mmol) in 1,4-dioxane (5 mL) was added aq. ammonia (0. 2 mL) at 0 °C andthe reaction mixture was stirred at room temperature for 3 h. The reaction mixture was concentrated and the residue was purified by column chromatography over silica gel using ethyl acetate/hexane (1 :9) mixture as eluent to give the title compound as an off white solid (0.010 g, 16%): 1H NMR (DMSO-d ₆ , 400 MHz) δ 1.59- 1.64 (m, 2H), 2.00-2.04 (m, 4H), 2.80-2.31 (m, 2H), 3.86 (t, J = 6.8 Hz, 2H), 7.03 (t, J = 7.6 Hz, 1H), 7.09 (d, J = 8 Hz, 1H), 7.19 (d, J = 7.2 Hz, 1H), 7.27 (t, J = 7.2 Hz, 1H), 7.37 (s, 1H), 7.54 (s, 1H), 9.41 (s, 1H);MS (ESI) m/z 427 (M+H) ⁺ ; HPLC purity: 94.65%.

Table II lists examples of compounds synthesised by the method of Scheme II. Table II

Figure 3 shows general reaction scheme ΙΠ for the synthesis of selectedl, 3-disubstituted phenothiazenes. Nucleophilic substitution of2-amino thiophenol/2-aminophenols (Ilia) with substituted aryl halides (IHb) followed by insituSmiles rearrangement yielded substituted phenothiazene/substituted phenoxazines (IIIc). Compounds IIIc were reducedusing Pd/C to yield the corresponding 1 -amino phenothiazenes/1 -amino phenoxazines (Hid). Compounds Hid were reacted with acid chlorides or acids to form corresponding amides Hie, which further deprotected to yield corresponding title compounds IIIg.Reductive amination of compound Hid with various aldehydes yielded correspondingw-alkylated phenothiazineslllf, which were further deprotected to give the corresponding title compounds IIIg.Further reductive amination of compound Illg, followed by deprotection gave ink. Detailed synthetic descriptions of some compounds syntehsised by the method of Scheme III provided below.

Compound 105: 3-Amino-iV-(3-(trifluoromethyl)-10H-phenothiazin-l- yl)cyclohexanecarboxamide

Step 1: l-Nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-chloro-l ,3-dinitro-5-(trifluoromethyl)benzene (25 g, 92.5 mmol) in water (200 mL) were added 2-aminobenzenethiol (10.4 g, 83.3 mmol), sodium hydroxide (11.1 g, 277.5 mmol) and the reaction mixture was heated at 80 °C for 12 h. The reaction mixture was cooled to room temperature, filtered, and thesolid obtained was washed with water followed by EtOH to give the title compound as a brown solid (25.0 g, 87%): ¹ H NMR (DMSO-d _6, 400 MHz) δ 6.97 (m, 1H), 7.02 (m, 1H), 7.09 (m, 2H), 7.65 (s, 1H), 8.01 (s, 1H), 9.84 (s, 1H); MS (ESI) «¾/z312 (M+H) ⁺ .

Step 2: 3-(Trifluoromethyl)-10H-phenothiazin-l -amine

To a stirred solution of l-nitro-3-(trifluoro methyl)- 10H-phenothiazine( 25 g, 801 mmol) in MeOH (250 mL) was added 10% Pd/C (50% wet, 5 g) and the reaction mixture was hydrogenated under ¾ gas (balloon) at room temperature for 16 h. The reaction mixture was filtered through celite and filtrate was concentratedto give the title compound as a light pink solid (20 g, 88%):1H NMR (DMSO-d _6, 400 MHz) δ 5.44 (s, 2H), 6.49 (s, 1H), 6.73 (s, 1H), 6.81(t, J = 7.2 Hz, 1H), 6.92 (d, J = 8 Hz, 1H), 6.92 (d, J = 7.6 Hz, 1H), 7.01 (t, J = 7.6 Hz, 1H), 7.81 (s, 1H); MS (ESI) «¾/z283 (M+H) ⁺ . Step 3: Tert-butyl (3-((3-(trifluoromethyl)-10H-phenothiazin- lyl)carbamoyl)cyclohexyl)carbamate

To a stirred solution of 3 -((tert-butoxycarbonyl)amino)cyclohexanecarboxylic acid(10.3 g, 42.55 mmol) in pyridine (25 mL) was added POCI ₃ (10 mL) at 0 °C and the reaction mixture was stirred for 15 min at 0 °C. A solution of (3-(trifluoromethyl)-10H-phenothiazin-l -amine (10 g, 35.46 mmol) in pyridine (25 mL) was added to the reaction mixture and stirring continued at room temperature for 1 h. The reaction mixture was concentrated and the residue was purified by column chromatography over silica gel using 20% ethyl acetate/hexane mixture as eluant to give the title compoundas a light yellow solid (6 g, 33%): ¹ H NMR (DMSO-d _6, 400 MHz) δ 1.09 (m, 1H), 1.18 (m, 4H), 1.30 (s, 9H), 1.37 (m, 2H), 1.97 (m, 2 H), 3.35(s, 2H), 3.55 (s, 2 H), 4.45 (bs, 1H), 6.79 (m, 1H), 6.84 (d,J = 6 Hz,2H), 6.96 (t, J = 7.2 Hz, 1H), 7.51 (d, J = 8 Hz, 1H),8.13 (s, 1H);MS (ESI) m/z506 (M+H) ^" .

Step 4: 3-Amino-/V-(3-(trifluoromethyl)-10H-phenothiazin-l-yl)cycloh exanecarboxamide

To a stirred solution of /er/-butyl (3-((3-(trifluoromethyl)-10H-phenothiazin-l - yl)carbamoyl)cyclohexyl)carbamate(6.8 g,13.4 mmol) in dichloromethane (60 mL) at 0 °C was added a solution of 4N HC1 in dioxane (20 mL)and stirring continued at room temperature for 12 h. The reaction mixture was concentratedand the residue was triturated with diethyl ether and n- hexane to give the title compound as an off-white solid (2 g, 37%): H NMR (DMSO-d ₆ , 400 MHz) 51.18- 1.22 (m, 4H), 1.75-1.78 (m, 2H), 1.84-1.86 (m, 1H),1.98- 2.07(d, J = 12.4 Ηζ,ΙΗ), 2.59 (s, 1 H), 3.45-3.48(m, 1H), 6.83-6.91 (m, 2H), 6.96 (d,J = 6.8 Hz ,1H), 7.02 (t, J = 7.6 Hz, 1H), 7.12 (s, 1H), 8.13 (s, 1H), 7.38 (s, 1H), 8.03 (s, 1H);MS (ESI) «¾/z408 (M+H) ⁺ ; HPLC purity: 99.65%.

Compound 133: iV-((3-Aminocyclohexyl)methyl)-3-(trifluoromethyl)-10H-pheno thiazin-l- amine

Step 1: l-Nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-chloro-l,3-dinitro-5-(trifluoromethyl)benzene (25 g, 92.5mmol) in water (200 mL) were added 2-aminobenzenethiol (10.4 g, 83.3 mmol), sodium hydroxide (11.1 g, 277.5 mmol) and the reaction mixture was heated at 80 °C for 12 h. The reaction mixture was cooled to room temperature, filtered, and thesolid obtained was washed with water followed by EtOH to give the title compound as a brown solid (25.0 g, 87%):1H NMR (DMSO-d _6, 400 MHz) δ 6.97 (m, 1H), 7.02 (m, 1H), 7.09 (m, 2H), 7.65 (s, 1H), 8.01 (s, 1H), 9.84 (s, 1H); MS (ESI) «¾/z312 (M+H) ⁺ .

Step 2: 3-(Trifluoromethyl)-10H-phenothiazin-l -amine

To a stirred solution of l-nitro-3-(trifluoromethyl)-10H-phenothiazine(25 g, 80.1 mmol) in MeOH (250 niL) was added 10% Pd/C (50% wet, 5 g) and the reaction mixture was hydrogenated with ¾ gas (balloon) at room temperature for 16 h. The reaction mixture was filtered through celite and filtrate was concentratedto give the title compoundas a light pink solid (20 g, 88%): ¹ H NMR (DMSO-d _6, 400 MHz) δ 5.44 (s, 2H), 6.49 (s, 1H), 6.73 (s, 1H), 6.81(t, J = 7.2 Hz, 1H), 6.92 (d, J = 8 Hz, 1H), 6.92 (d, J = 7.6 Hz, 1H), 7.01 (t, J = 7.6 Hz, 1H), 7.81 (s, 1H); MS (ESI) «¾/z283 (M+H) ⁺ .

Step 3: 7eri-butyl(3-(((3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)methyl)cyclohexyl)carbamate

To a stirred solution of 3-(trifluoromethyl)-10H-phenothiazin-l -amine (10 g, 35.46 mmol), tert- butyl (3-formylcyclohexyl)carbamate (12 g, 54.05 mmol) in MeOH (100 mL) was added AcOH (2.5 mL) at 0 °C and the reaction was stirred for 1 h at 0 °C. NaCNBH ₃ (1 1 g, 177.3 mmol) was added to the reaction mixture at 0 °C and was stirred at room temperature for 12 h. The reaction mixture was evaporated, the residue was diluted with EtOAc, and washed with ag.NaHCC solution. The organic layer was dried over anhydrous Na ₂ S0 ₄ , concentrated, and the residue was purified by column chromatography over silica gel using 20% ethyl acetate/hexane as eluant to give the title compound as a light yellow solid (11 g, 62%): ¾ NMR (DMSO-d _6, 400 MHz) δ 0.77-0.89 (m, 2H), 1.02-1.04 (m, 1H), 1.19-1.25 (m, 2H), 1.35 (s, 9H), 1.63-1.78 (m, 4H), 1.95- 1.98 (m, 1H), 3.14-3.27 (m, 3H), 4.01-4.05 (m, 1H), 5.42 (bs, 1H), 6.50 (s, 1H), 6.53 (s, 1H), 6.72 (d, J = 6.8 Hz, 1H), 6.81 (t, J = 7.2 Hz, 1H), 6.87 (d, J = 8 Hz, 1H), 6.93 (d, J = 7.2 Hz, 1H), 7.02(t, J = 7.6 Hz, 1H), 7.99 (s, 1H);MS (ESI) m/z 494 (M+H) ⁺ .

Step 4: iV-((3-Aminocyclohexyl)methyl)-3-(trifluoromethyl)-10H-pheno thiazin-l-amine

To a stirred solution of tert-butyl (3-((3-(trifluoromethyl)-10H-phenothiazin-l- yl)carbamoyl)cyclohexyl)carbamate(10 g,20.24 mmol) in dichloromethane (75 mL) at 0 °C was added a solution of 4N HC1 in dioxane (15 mL)andwas stirredat room temperature for 12 h. The reaction mixture was concentratedand the residue was triturated with diethyl ether followed by hexane. The crude product was diluted with water, basified with ag.NaHCC solution, extracted with EtOAc, organic layer is dried over anhydrous Na ₂ S0 ₄ ,concentrated, and the crude product was triturated with acetonitrile to get the title compoundas pale yellow solid (5 g, 63%): ¹ H NMR (DMSO-de _, 400 MHz) δ 0.65-0.94 (m, 3H), 1.13-1.26 (m, 1H),1.36-1.90 (m, 6H), 1.93- 1.96 (m, 1 H), 2.84-2.98 (m, 2H), 5.53 (s, 1H), 6.48 (s, 1H), 6.52 (s, 1H), 6.80 (t, J = 7.6 Hz, 1H), 6.91 (q, J = 7.6 Hz, 2H),7.02 (t, J = 7.6 Hz, 1H), 8.06 (s,lH): MS (ESI) m/z 394 (M+H) ⁺ ; HPLC purity: 99.78%.

Compound 140: iV-(l-(2-Aminoethyl)piperidin-4-yl)-3-(trifluoromethyl)-10H- phenothiazin- 1-amine

Step 1: l-Nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-chloro-l ,3-dinitro-5-(trifluoromethyl)benzene (25 g, 92.5 mmol) in water (200 mL) were added 2-aminobenzenethiol (10.4 g, 83.3 mmol), sodium hydroxide (11.1 g, 277.5 mmol) and the reaction mixture was heated at 80 °C for 12 h. The reaction mixture was cooled to room temperature, filtered, and thesolid obtained was washed with water followed by EtOH to give the title compound as a brown solid (25.0 g, 87%): ¹ H NMR (DMSO-d _6, 400 MHz) δ 6.97 (m, 1H), 7.02 (m, 1H), 7.09 (m, 2H), 7.65 (s, 1H), 8.01 (s, 1H), 9.84 (s, 1H); MS (ESI) «¾/z312 (M+H) ⁺ .

Step 2: 3-(Trifluoromethyl)-10H-phenothiazin-l -amine

To a stirred solution of l-nitro-3-(trifluoromethyl)-10H-phenothiazine(25 g, 80.1 mmol) in MeOH (250 mL) was added 10% Pd/C (50% wet, 5 g) and the reaction mixture was hydrogenated under ¾ gas (balloon) at room temperature for 16 h. The reaction mixture was filtered through celite and filtrate was concentratedto give the title compound as a light pink solid (20 g, 88%):1H NMR (DMSO-d _6, 400 MHz) δ 5.44 (s, 2H), 6.49 (s, 1H), 6.73 (s, 1H), 6.81(t, J = 7.2 Hz, 1H), 6.92 (d, J = 8 Hz, 1H), 6.92 (d, J = 7.6 Hz, 1H), 7.01 (t, J = 7.6 Hz, 1H), 7.81 (s, 1H); MS (ESI) «¾/z283 (M+H) ⁺ .

Step 3: Tert-butyl 4-((3-(trifluoromethyl)-10H-phenothiazin-l-yl)amino)piperidi ne-l carboxylate

To a stirred solution of 3-(trifluoromethyl)-10H-phenothiazin-l-amine (10 g, 35.4 mmol)in DCE (250 mL) was added tert-butyl 4-oxopiperidine-l -carboxylate(8.4 g, 42.5 mmol) and the reaction mixture was stirred for 1 h at room temperature. Na(OAc) ₃ BH (11.26 g, 53.13 mmol) was added at room temperature and stirring was continued at room temperature for 16 h. The reaction mixture was diluted with ag.NaHCC solution and extracted with DCM. The organic layer was dried over Na ₂ S0 ₄ , concentrated, and the residue was as such take for the next stepwithout further purification (13 g, crude): MS (ESI) m/z 466 (M+H) ⁺ .

Step 4: iV-(Piperidin-4-yl)-3-(trifluoromethyl)-10H-phenothiazin-l-a mine

To a stirred solution of tert-butyl 4-((3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)piperi dine- 1-carboxy late (13 g, 35.6 mmol) in dichloromethane (150 mL) was added 4N HC1 in dioxane (20 mL) at 0 °C and stirred at room temperature for 5 h. The reaction mixture was concentrated, residue was diluted withaqueous sodium bicarbonate solution, extracted with dichloromethane, and concentrated. The residue was purified over silica gel using 8% MeOH/DCM as eluant to give the title compound as black solid (6.58 g, 66%);MS (ESI) m/z366 (M+H) ⁺ .

Step 5: Tert-butyl (2-(4-((3-(trifluoromethyl)-10H-phenothiazin-l-yl)amino)pipe ridin-l- yl)ethyl)carbamate

To a stirred solution of N-(piperidin-4-yl)-3-(trifluoromethyl)-10H-phenothiazin-l-am ine(5.58 g, 15.2 mmol) in acetonitrile (70 mL) were added tert-butyl (2-bromoethyl)carbamate (4.1 g, 18.3 mmol), potassium carbonate (6.3 g, 45.8 mmol) and was stirred at 70 °C for 16 h. The reaction mixture was concentrated, diluted with water,and extracted with ethyl acetate. The organic layer was washed with brine solution, dried over anhydrous sodium sulphate, andconcentrated. The residue was purified by column chromatography over silica gel using 8% methanol/dichloromethane mixture as eluant to give the title compound asbrown solid(4 g, 52 %);MS (ESI) m/z 509 (M-H) ⁺ .

Step 6: /V-(l-(2-Aminoethyl)piperidin-4-yl)-3-(trifluoromethyl)-10H- phenothiazin-l-amine

To a stirred solution of /er/-butyl(2-(4-((3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)piperidin-l-yl)ethyl)carbamate (7.5 g, 14.7 mmol)) in dichloromethane (100 mL) at 0 °C was added 4M HQ in dioxane (10 mL) and the reaction was stirred at room temperature for 5 h. The reaction mixture was concentrated and washed with pentane. The crude product was dissolved in water, basified with ag.NaHCC solution, filtered, and solid was triturated with diethyl ether followed by w-pentane to get the desired product N-(l-(2-aminoethyl)piperidin-4- yl)-3-(trifluoromethyl)-l OH-phenothiazin-1 -amine as brown solid (3.9 g, 65%): Ή NMR (DMSO-d _6, 400 MHz) δ 1.41 -1.48 (m, 4H), 1.90-1.93 (d, J= 12 Hz, 2 H), 2.063 (t, J=12 Hz, 2H), 2.30 (t, J=4 Hz, 2H), 2.58-2.61 (m, 2H), 2,82-2.84 (d, J=8 Hz, 2H), 5.26-5.28 (d, J=8 Hz, 1H), 6.54 (s, 1H), 6.58 (s, 1H), 6.80-6.82 (m, 1H), 6.84-6.88 (m, 1H), 6.90-6.95 (m, 1H), 7.01-7.05 (m, 1H), 8.02 (s, 1H);MS (ESI) m/z409 (M+H) ⁺ ; HPLC purity: 99.10%.

Table III lists examples of compounds synthesised by the method of Scheme III. Table III

Scheme IV; Synthesis of 3^V-10-disubstituted phenothiazenes

Figure 4 shows general synthetic scheme IV for the synthesis of selected3,N-10-Disubstituted phenothiazenes. Nucleophilic substitution of 2-amino-5-bromobenzenethiol (TVa) with 1 -chloro- 2-nitrobenzene (IVb) resulted in compound IVc. N-Formylation followed bySmiles rearrangement of compound IVc yielded 3-bromo phenothiazene (IVe).N-alkylation of compound IVe with alkylbromides using NaH yielded N-10- alkylatedphenothiazene(rVf).Neucleophilic substitution of compound IVf followed by reduction, and protection of the resulting amine gave compound IVg.Parallelly, reaction with amines compound IVf resulted in compound IVh. Suzuki coupling of compounds IVg&IVh with arylborane compounds yielded correspoinding 3-aryl phenothiazenes (IVj&IVi). Deprotection of compoundsIVj&rVi gives title compoundsIVl&IVk. Finally, reaction of IVk with acylchlorides or alkylhalides resulted in the corresponding title compounds IV1.

Detailed synthesis descriptions of some compounds synthesised by the method of Scheme IV is provided below.

Compound 111: 3-(3-(lH-Indol-2-yl)-10H-phenothiazin-10-yl)-iV^V-dimethylpr opan-l- amine

Step 1: 2-((4-Bromo-2-nitrophenyl)thio)aniline

To a stirred solution of 1 ,4-dibromo-2-nitrobenzene (50 g, 179 mmol) in EtOH (500 mL) were added 2-aminobenzenethiol (23 mL, 215.16 mmol), KOH (8.6 g, 215.16 mmol) and the reaction was stirred at room temperature for 16 h. The precipitated solid was filtered, washed with n- hexane, dried under vacuum to give the title compound as yellow solid(65 g, quantitative) :1H NMR (CDC1 _3, 400 MHz) δ 5.53 (s, 2H), 6.59-6.67 (m, 2H), 6.80 (d, J = 8.0 Hz, 1H), 7.23 (t, J = 7.6.0 Hz, 1H), 7.31 (d, J= 7.6 Hz, 1H), 7.73-7.75 (m, 1H), 8.35-8.36 (m, 1H). Step 2: iV-(2-((4-bromo-2-nitrophenyl)thio)phenyl)formamide

A mixture of 2-((4-bromo-2-nitrophenyl)thio)aniline(60 g) and formic acid (300 mL) was heated at 100 °C for 16 h. The reaction mixture was evaporated, the crude was poured on crushed ice, and the solid obtained was filtered, dried under vacuum to get the title compound as yellow solid(50 g, 92%): ¹ H NMR (CDC1 _3, 400 MHz) δ 6.52 (d, J = 7.2 Hz, 1H), 7.257.29 (m, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.67(d, J = 7.6 Hz, 1H), 7.73 (d, J = 7.6 Hz, 1H), 8.15 (s, 1H), 8.33 (d, J = 8 Hz, 1H), 8.40 (s, 1H), 9.72 (s, 1H).

Step 3: 3-Bromo-10H-phenothiazine

To a stirred solution of N-(2-((4-bromo-2-nitrophenyl)thio)phenyl)formamide(50 g, 142 mmol) in acetone (500 mL) was added KOH (25 g, 426 mmol) and the reaction mixture was stirred at 80 °C for 16 h. The reaction mixture was evaporated, the crude was suspended in water, and stirred for 5 h. The solid was filtered and dried under vacuum to get the title compound as light brown solid(40 g, quantitative) :MS (ESI) «¾/z278 (M+2H) ⁺ .

Step 4: 3-Bromo-10-(3-chloropropyl)-10H-phenothiazine

To a stirred solution of 3-bromo-10H-phenothiazine(18 g, 64.72 mmol) in DMF (100 mL) was added sodium hydride (3.8 g, 97.08 mmol) at 0 °C in small portions and the reaction mixture was stirred at 0 °C for 15 mins. To the reaction mixture was added 3-bromo chloropropane (12.2 g, 77.66 mmol) at 0 °C and stirring continued at room temperature for 1 h. The reaction was quenched with a</.NH ₄ Cl solution, extracted with EtOAc, organic layer is washed with brine, dried over anhydrous sodiumsulphate, and concentrated. The crude product (20 g) was as such taken for the next step without further purification: MS (ESI) m/z355 (M+2H) ⁺ .

Step 5:3-(3-Bromo-10Hphenothiazin-10-yl)-iV^V-dimethylpropan-l-am ine

To a stirred solution of 3-bromo-10-(3-chloropropyl)-10H-phenothiazine(15 g, 42.28 mmol) in DMF (150 mL) were added 2M solution of N,N-dimethyl amine in THF (4.7 mL, , 84.57 mmol),potassium phosphate (26.9 g, 126.84 mmol) and the reaction mixture was stirred at 80 °C for 12 h. The reaction mixture was diluted with water, and extracted with EtOAc. The organic layer was washed with ice-cold water followed by brine solution, dried over sodium sulphate, and concentrated. The crude product was purified over silica gel using5% MeOH/DCM as eluant to give title compound as brown hquid(8 g, 53%): 1H NMR (DMSO _, 400 MHz) δ 1.69-1.76 (m, 2H), 2.05 (s, 6H), 2.23-2.26 (m, 2H), 3.83-3.86 (m, 2H), 6.91-6.94 (m, 2H), 7.0 (d, J = 8 Hz, 1H), 7.12 (d, J= 7.6 Hz, 1H), 7.18-7.20 (m, 1H), 7.30-7.33 (m, 2H); MS (ESI) m/z365 (M+2H) ⁺ .

Step 6: 7eri-butyl-2-(10-(3-(dimethylamino)propyl)-10H-phenothiazin- 3-yl)-lH-indole-l- carboxylate

To a solution of 3-(3-bromo-10H-phenothiazin-10-yl)-N,N-dimethylpropan-l-amin e( 3 g, 8.25 mmol) in a mixture of DME/water (50/5 mL) were added potassium carbonate (3.4 g, 24.75 mmol), (l-(tert-butoxycarbonyl)-lH-indol-2-yl)boronic acid (3.2 g, 12.38 mmol) and reaction mixture was purged with nitrogen for 10 mins. To the reaction mixture was added bis(triphenylphosphine)palladium(II) dichloride (0.57 g, 0.82 mmol), was purged with nitrogen for 10 mins, and refluxed for 16 h. The reaction mixture was filtered through celite, filtrate was diluted with EtOAc, washed with water, dried over sodium sulphate, and concentrated. The crude product was purified over silica gel using 4% MeOH/DCM as eluant to get title compound as brown viscous liquid (2 g, 48%): ¹ H NMR (DMSO _, 400 MHz) δ 1.28 (s, 9H), 1.71-1.81 (m, 2H), 2.09 (s, 6H), 2.29-2.33 (m, 2H), 3.91-3.95 (m, 2H), 6.67 (s, 1H), 6.93 (t, J = 7.6 Hz, 1H), 7.05 (t, J = 8.8 Hz, 2H), 7.13-7.30 (m, 6H), 7.56 (d, J = 7.6 Hz, 1H), 8.04 (d, J = 8.4 Hz, 1H); MS (ESI) m/z 500 (M+H) ⁺ .

Step 7: 3-(3-(lH-Indol-2-yl)-10H-phenothiazin-10-yl)-/V^V-dimethylpr opan-l-amine

To a solution of /er/-butyl-2-(10-(3-(dimethylamino)propyl)-10H-phenothiazin- 3-yl)-lH-indole- 1 -carboxylate (3 g, 6 mmol) in MeOH (30 mL) was added potassium carbonate (2.5 g, 18 mmol) and the reaction mixture was heated at 70 °C for 12 h. The reaction mixture was evaporated, residue was diluted with water, extracted with ethyl acetate,organic layer was dried over sodium sulphate, and concentrated. The crude product was purified over silica gel using 5% MeOH/DCM as eluant to afford 3-(3-(lH-indol-2-yl)-10H-phenothiazin-10-yl)-N,N- dimethylpropan-1 -amine as off-white solid (1.3 g, 54%): ¹ H NMR (DMSO-d _6, 400 MHz) δ 1.94 (s, 6H), 2.31 (s, 2H), 2.66 (s, 2H), 3.96 (s, 2H), 6.80 (s, 1H), 6.96-6.97 (m, 2H), 7.04-7.08 (m, 2H), 7.12 (d, J = 8.0 Hz, 1H), 7.19-7.24 (m, 2H), 7.35 (d, J = 8.0 Hz, 1H), 7.48 (d, J = 8.0 Hz, 1H), 7.66-7.69 (m, 2H), 11.40 9s, 1H); MS (ESI) m/z 400 (M+H) ⁺ . Example 150: 3-(3-(lH-Indol-2-yl)-10H-phenothiazin-10-yl)propan-l-amine

Step 1: 2-((4-Bromo-2-nitrophenyl)thio)aniline

To a stirred solution of 1 ,4-dibromo-2-nitrobenzene (50 g, 179 mmol) in EtOH (500 mL) were added 2-aminobenzenethiol (23 mL, 215.16 mmol), NaOH (8.6 g, 215.16 mmol) and the reaction mixture was stirred at room temperature for 16 h. The precipitated solid was filtered, washed with ft-hexane, and dried under vacuum to give the title compound as yellow solid(65 g, quantitative) :1H NMR (CDC1 _3, 400 MHz) δ 5.53 (s, 2H), 6.59-6.67 (m, 2H), 6.80 (d, J = 8.0 Hz, 1H), 7.23 (t, J= 7.6.0 Hz, 1H), 7.31 (d, J= 7.6 Hz, 1H), 7.73-7.75 (m, 1H), 8.35-8.36 (m, 1H).

Step 2:iV-(2-((4-Bromo-2-nitrophenyl)thio)phenyl)formamide

A mixture of 2-((4-bromo-2-nitrophenyl)thio)aniline(60 g, 185mmol) and formic acid (300 mL) was heated at 100 °C for 16 h. The reaction mixture was evaporated, the crude was poured on crushed ice, precipitated solid was filtered, and dried to get the title compoundas yellow solid(50 g, 92%): ¹ H NMR (CDC1 _3, 400 MHz) δ 6.52 (d, J = 7.2 Hz, 1H), 7.257.29 (m, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.67(d, J = 7.6 Hz, 1H), 7.73 (d, J = 7.6 Hz, 1H), 8.15 (s, 1H), 8.33 (d, J = 8 Hz, 1H), 8.40 (s, 1H), 9.72 (s, 1H). Step 3:3-Bromo-10H-phenothiazine

To a stirred solution of N-(2-((4-bromo-2-nitrophenyl)thio)phenyl)formamide(50 g, 142 mmol) in acetone (500 mL) was added KOH (25 g, 426 mmol) and thereaction mixture was stirred at 80 °C for 16 h. The reaction mixture was evaporated, the residue was suspended in water, and was stirred for 5 h. The precipitated solid was filtered, dried to get the title compound as light brown solid(40 g, quantitative):MS (ESI) «¾/z278 (M+2H) ⁺ .

Step 4 :3-Bromo- 10-(3-chloropropyl)- 1 OH-phenothiazine

To a stirred solution of 3-bromo-10H-phenothiazine(18 g, 64.72 mmol) in DMF (100 mL) was added sodium hydride (3.8 g, 97.08 mmol) at 0 °C in small portions and the reaction mixture was stirred at 0 °C for 15 mins. To the reaction mixture was added 3-bromo chloropropane (12.2 g, 77.66 mmol) at 0 °C and stirring continued at room temperature for 1 h. The reaction mixture was quenched with a</.NH ₄ Cl solution, extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous sodiumsulphate, and concentrated. The crude product (20 g) was as such taken for the next step without further purification: MS (ESI) m/z355 (M+2H) ⁺ .

Step 5: 10-(3-Azidopropyl)-3-bromo-10H-phenothiazine

To a stirred solution of 3-bromo-10-(3-chloropropyl)-10H-phenothiazine (10 g, 28.19 mmol) in DMSO (100 mL) was added sodium azide (10.9 g, 161.5 mmol) and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with ice, extracted with EtOAc, the organic layer was dried over sodium sulphate, and concentrated. The crude product was purified over silica gel using 100% hexane as eluant to give the desired productas viscous solid (9 g, 88%):1H NMR (DMSO _, 400 MHz) δ 1.84-1.91 (m, 2H), 3.39-3.43 (m, 2H), 3.90-3.94 (m, 2H), 6.93-6.98 (m, 2H), 7.05 (d, J = 8 Hz, 1H), 7.12-7.22 (m, 2H), 7.32-7.34 (m, 2H); MS (ESI) m/z365 (M+2H) ⁺ .

Step 6:3-(3-Bromo-10H-phenothiazin-10-yl)propan-l-amine

To a stirred solution of 10-(3-azidopropyl)-3-bromo-10H-phenothiazine (9 g, 24.86 mmol) in a mixture of THF (100 mL) and H ₂ 0 (20 mL) was added triphenylphosphine (13 g, 49.72 mmol) and was stirred at room temperature for 16 h. The reaction mixture was diluted with water, extracted with ethyl acetate, the organic layer is dried over sodium sulphate, and concentrated to get the desired crude product which was taken for next step without further purification(9 g):MS (ESI) m/z336 (M,M+2H) ⁺ .

Step 7: Tert-but i (3-(3-bromo-10H-phenothiazin-10-yl)propyl)carbamate

To a stirred solution of 3-(3-bromo-10H-phenothiazin-10-yl)propan-l-amine( 9 g, 25.33 mmol) in dichloromethane (100 niL) were added Et ₃ N (7.4 mL, 50.6 mmol), (Boc) ₂ 0 (11.6 mL, 50.6 mmol) at 0 °C and was stirred at room temperature for 2 h. The reaction mixture was concentrated andthe crude product was purified over silica gel using 40% EtOAc in hexane as eluant to get the desired product as brown viscous liquid(9 g, 78%): ¹ H NMR (DMSO _, 400 MHz) δ 1.74-1.78 (m, 2H), 2.99-3.01 (m, 2H), 3.80-3.84 (m, 2H), 6.79-6.85 (m, 1H), 6.90-7.02 (m, 3H), 7.12-7.19 (m, 2H), 7.30-7.34 (m, 2H); MS (ESI) m/z436 (M+2) ⁺ .

Step 8: Tert-but i 2-(10-(3-((tert-butoxycarbonyl)amino)propyl)-10H-phenothiazi n-3-yl)- lH-indole-l-carboxylate

To a stirred solution of tert-butyl (3-(3-bromo-10H-phenothiazin-10-yl)propyl)carbamate ( 2 g, 4.39 mmol) and (l-(/er/-butoxycarbonyl)-lH-indol-2-yl)boronic acid (1.3 g, 5.27 mmol) in a mixture of DME/Water (55/5 mL) was added potassium carbonate (0.9 g, 6.58 mmol) and reaction mixture was purged with nitrogen for 10 mins. To the reaction mixture was added bis(triphenylphosphine)palladium(II) dichloride (0.3 g, 0.43 mmol), was purged with nitrogen for 10 mins, and refluxed for 12h. The reaction mixture was filtered through celite, filtrate was diluted with EtOAc, washed with water followed by brine. The organic layer was dried over sodium sulphate, and concentrated. The crude was purified over silica gel using 15% EtOAc/hexane as eluant to give title compound as brown viscous liquid(1.6 g, 63 %): MS (ESI) m/z512 (M+H) ⁺ . Step 9:3-(3-(lH-Indol-2-yl)-10H-phenothiazin-10-yl)propan-l-amine

To a stirred solution of tert-butyl 2-(10-(3-((ieri-butoxycarbonyl)amino)propyl)-10H- phenothiazin-3-yl)-lH-indole-l-carboxylate (4 g, 70 mmol) in dichloromethane (60 mL) was added TFA (12 mL) at 0 °C and was stirred at room temperature for 16 h. The reaction mixture was concentrated, residue was basified with saturated sodium bicarbonate solution, extracted with dichloromethane, the organic layer was dried over sodium sulphate, and concentrated. The residue was washed with diethyl ether led to 3-(3-(lH-indol-2-yl)-10H-phenothiazin-10- yl)propan-l -amine as pale yellowsohd (1.9 g, 73%): ¹ H NMR (DMSO-i/ _6, 400 MHz) δ 1.78-1.81 (m, 2H), 2.67-2.71 (m, 2H), 3.93-3.96 (m, 2H), 6.77 (s, 1H), 6.93-6.98 (m, 3H), 7.02-7.21 (m, 6H), 7.33 (d, J = 8 Hz, 1H), 7.56 (d, J = 7.6 Hz, 1H), 7.62-7.66 (m, 2H), 11.37(s, 1H); MS (ESI) m/z 372 (M+H) ⁺ ; HPLC purity: 99.45%.

Table IV lists some examples of compounds synthesised by the method of Scheme TV. Table IV

Figure 5 shows general synthetic scheme V for the synthesis of selected 1,3-disubstituted phenothiazenes.Nucleophilic substitution of 2-aminothiophenol (Va) with aryl halides (Vb) followed by insituSmiles rearrangement using NaOH gives 1 ,3-disubstituted phenothiazene (Vc). Acid-amine coupling of compound Vc with distinct amines resulted in the corresponding amides Vd. Reducing the N0 ₂ group of Vd using Pd/C resulted in the corresponding amine compound Ve. Amide formation of compound Ve using corresponding acids or acid chlorides followed by deprotection of amine group resulted in title compounds Vh. Parallel, reductive amination of compound Ve with carbonyl compound followed by deprotection resultedin title compound Vh.

Some examples of compounds synthesised by the method of Scheme V are listed in Table V.

Table V

Figure 6 shows general synthetic scheme VI for the synthesis of selected 3, N-10-Disubstituted azaphenothiazenes. Nucleophilic substitution of substituted amino thiophenols (Via) with 2- chloro-3-nitro pyridine (VIb) gives compound Vic. Acetylation followed by Smiles rearrangement of Vic yielded compound Vie. Deprotection of compound Vie using HCl gives 3-haloazaphenothiazenes Vlf. Alkylation of compound Vlf using alkyl chlorides NaH followed by Suzuki coupling with arylboronic acid resulted title compounds Vlh. A detailed synthetic descrription of a compound synthesised by the method of Scheme VI is provided below. Compound 135: 3-(7-(lH-Indol-2-yl)-10H-benzo[6]pyrido[2,3-e] [l,4]thiazin-10-yl)-iV - dimethylpropan-l-amine

Step 1: 4-Bromo-2-((3-nitropyridin-2-yl)thio)aniline

To a stirred solution of 4-bromo-2 amino-benzenethiol (2.0 g, 9.8 mmol) and 2-chloro3-nitro- pyridine (2.0 g, 12.7 mmol) in ethanol was added sodium hydroxide (1.2 g, 29.4 mmol) at r.t. and stirring continuedat r.t for 4 h. Reaction mixture was filtered, solid waswashed with DM water, and dried completly to offered the desired product as yellow solid (1.3 g, 42%):NMR (DMSO-d _6, 400 MHz) δ 5.55 (s, 2H), 6.68 (d, J= 8.8 Hz, 1H) 7.27 (d, J= 1.6 Hz, 1H), 7.29-7.42 (m, 2H), 8.59 (d, J= 8.0 Hz, 2H);MS (ESI) m/z 326 (M+H) ⁺ .

Step 2 : iV-(4-Bromo-2-((3-nitropyridin-2-yl)thio)phenyl)acetamide

To a stirred solution of 4-bromo-2-((3-nitropyridin-2-yl)thio)aniline (1.8 g, 5.52 mmol) in pyridine (5mL) was added acetic anhydride (2.8mL, 27.6 mmol) at 0°C, and stirring continued at r.t for 3h. Reaction mixture was concentrated, diluted with DM water (lOOmL), extracted with EtOAc, dried the organic layer over sodium sulphate, and concentrated.Product was washed with ether to get the titlecompoundas pale yellow solid(2 g,97%): MS (ESI) m/z 368 (M+H) ⁺ .

Step 3: 7-Bromo-10H-benzo[6]pyrido[2^-e] [l,4]thiazine

To a stirred solution of N-(4-bromo-2-((3-nitropyridin-2-yl)thio)phenyl)acetamide(2.0 g, 5.43 mmol) in acetone (40 mL) was added KOH (0.9 g, 16.3 mmol) and the reaction mixture was heated at 60 °C for 16 h. The reaction mixture was cooled to room temperature, concentrated, and the residue was purified by column chromatography over silica gel using 30% ethyl acetate/hexane mixture as eluant to give the title compound as an dark solid (1.4 g, 82%): MS (ESI) m/z 279 (M+H) ⁺ Step 4: 3-(7-Bromo-10H-benzo[b]pyrido[2,3-e] [l,4]thiazin-10-yl)-iV,iV-dimethylpropan-l- amine

To a stirred solution of sodium hydride (0.350g, 7.16 mmol) in DMF (10 mL) at 0 °C was added a solution of 7-bromo-10H-benzo[6]pyrido[2,3-e][l ,4]thiazine(1.0 g, 3.58 mmol) in DMF and stirring was continuedfor 20 min.To the reaction mixture3-chloro-N,N-dimethylpropan-l -amine hydrochloride (0.1.12 g, 7.16 mmol) was added and the reaction mixture was heated at 65°C for 16h. The reaction mixture was cooled to room temperature, concentrated, and the residue was purified by column chromatography over silica gel using 5%methanol/dichloromethane mixture as eluant to afford title compound as gummy material(0.55 g, 45%): MS (ESI) m/z 364(M+H) ⁺ . Step 5: Tert-but i 2-(10-(3-(dimethylamino)propyl)-10H-benzo[b]pyrido[2,3-e] [l,4]thiazin- 7-yl)-lH-indole-l-carboxylate

To a solution of 3-(7-bromo-10H-benzo[¾]pyrido[2,3-e][l ,4]thiazin-10-yl)-N _r N-dimethylpropan- 1 -amine (0.30 g, 0.824 mmol) in DME/Water (6/2 mL) is added potassium carbonate (0.220 g, 1.64 mmol) and (l-(tert-butoxycarbonyl) lH-indol-2-yl)boronic acid (0.320 g, 0.1.23 mmol) and purged with nitrogen for 10 min.followed by added bis(triphenylphosphine)palladium(II) dichloride (0.057 g, 0.082 mmol). Reaction mixture was again purged with nitrogen for 10 mins and refluxed for 12h. The reaction mixture wais filtered through diatomaceous earth. The filtrate is dried over sodium sulphate and concentrated. The crude product was purified on silica column using5% methanol in dichloromethane as eluant to afford the titlecompoundas gummy matenal(0.120 g, 30%): 1H NMR (DMSO-d _6, 400 MHz) δ 1.16 (s, 4H), 1.28 (s, 9H), 1.80-1.87 (m, 2H), 2.23 (s, 5H), 2.31 -2.49 (m, 2H), 4.03-4.12 (m, 2H),6.70(s, 1Η),6.86-6.89 (m, 2H), 7.08 (d, J = 8.8 Hz, 1H), 7.17(d, J = 2.0 Hz, 1H), 7.23-7.32 (m, 3H), 7.43-7.45 (m, 1H), 7.57 (d, J = 7.2 Hz, 1H), 8.02-8.07 (m, 2H); MS (ESI) m/z 501 (M+H) ⁺ .

Step 6:3-(7-(lH-Indol-2-yl)-10H-benzo[6]pyrido[2,3-e] [l,4]thiazin-10-yl)-/V^V- dimethylpropan-l-amine

To a solution of tert-butyl 2-(10-(3-(dimethylamino)propyl)-10H-benzo[6]pyrido[2,3- e][l,4]thiazin-7-yl)-lH-indole-l -carboxylate (0.3 g, 0.599mmol) in dichloromethane (10 mL) is added 4N HQ in dioxane (1.0 mL) and reaction mixture was stirred for 4 h at room temperature. Reaction mixture was basified with solid NaHCC , extracted with dichloromethane, organiclayer is washed with saturated NaHCC solution, dried over sodium sulphate, and concentrated . The crude productwas purified bysilica gel column chromatographyusinglO% methanol in dichloromethane as eluant to afford the title compound as brown solid (0.040 g, 44%): ^X H NMR (DMSO 400 MHz) 50.84 (bs , H), 1.22(s, 2H), 1.85-1.89 (m, 2H), 2.18 (s, 6H), 2.31 -2.49 (m, 2H), 4.08 (t, J= 7.2Hz, 2H), 6.81(s, 1H), 6.86-6.89 (m,lH), 6.96(t, J= 7.6Hz, 1H), 7.04-7.12 (m, 2 H), 7.35(d, J= 8.4Hz, 1H), 7.47 (t, J= 8.0Hz, 2H), 7.58(s, lH),7.65(d, J= 8.8Hz, 1H), 8.02 (d, J= 3.6Hz, 1H), 11.39 (s, 1H);MS (ESI) m/z 401.2 (M+H) ^" ; HPLC purity: 99.2%. Some examples of compounds synthesised by the method of Scheme VI are listed in Table VI.

Figure 7 shows general synthetic scheme VII for the synthesis of selected 3, N-10-disubstituted phenothiazenes. Formylation of phenothiazene (Vila) at C-3 position usingurotropine yielded 3- formyl phenothiazene (Vllb). N-10 Alkylation of compound Vllb with alkyl bromide using NaHgivescompound VIIc. Reaction of compound VIIc with amine resulted in compound Vlld, which is further reacted with aryl diamine or aryl aminothiol to yield title compounds Vile.

A detailed synthetic description of a compound synthesised by the method of Scheme VII is provided below.

Compound 143: 3-(3-(Benzo[i/]thiazol-2-yl)-10H-phenothiazin-10-yl)-iV,iV-d imethylpropan- 1-amine

Step 1: 10H-Phenothiazine-3-carbaldehyde

To a solution of 1 OH-phenothiazine (3.0 g, 15.06 mmol) in acetic acid (25 mL) was added hexamethylenetetramine (3.15 g, 22.59 mmol) in a microwave vial and subjected to microwave irradiation. Reaction mixture was poured into water, neutralized with Na ₂ C03,extracted with EtO Ac. Organic layer was dried over sodium sulphateand concentrated. The crude productwas purified by biotage purifier with 40% ethyl acetate in hexane as eluent to afford the desired product as a yellow solid (0.33 g, 10%): 1H NMR (CDC1 _3, 400 MHz) δ 6.07 (s, 1H), 6.52-6.57 (m, 2H), 6.84-6.88 (m, 1H), 6.94 (4, J = 7.6 Hz, 1H), 6.99 (t, J = 7.6 Hz, 1H), 7.44 (s, 1H), 7.47 (d, J= 8.0 Hz, 1H), 9.71 (s, 1H); MS (ESI) m/z 228 (M+H) ⁺

Step 2: 10-(3-Chloropropyl)-10H-phenothiazine-3-carbaldehyde

To a solution of NaH (0.078 g, 1.95 mmol) in N V-dimethyl formamide (8.0 niL) was added 10H-phenothiazine-3-carbaldehyde ( 0.3 g,1.31 mmol) at 0 °C and reaction mixture was stirred for 0.5 h at room temperature. To the reaction mixture was addedl -bromo-3-chloro-propane (0.247 g,1.57 mmol) at 0 °C and stirring continued at room temperature for 3 h. The reaction mixture was quenched with ice and extracted with EtO Ac. Organic layer was washed with water, saturated NaHC03 solution,brine, dried over sodium sulphate, and concentrated. The crude productwas purified by biotage purifier with 3% ethyl acetate in hexane as eluent to afford the title compound as a viscous liquid (0.15 g, 40%): 1H NMR (DMSO-d _6, 400 MHz) δ 1.1 1 -1.14 (m, 2H), 3.72-3.75 (m, 2H), 4.10-4.13 (m, 2H), 7.02 (t, J = 7.6 Hz, 1H), 7.14 (d, J = 7.6 Hz, 1H), 7.19 (d, J = 7.6 Hz, 1H), 7.24 (t, J = 8.0 Hz, 2H), 7.62 (s, 1H), 7.73 (d, J = 8.4 Hz, 1H), 9.79 (s, 1H); MS (ESI) m/z 304 (M+H) ⁺ . Step 3: 10-(3-(Dimethylamino)propyl)-10H-phenothiazine-3-carbaldehyd e

To a stirred solution of 10-(3-chloropropyl)-10H-phenothiazine-3-carbaldehyde (0.15 g, 0.495 mmol) in DMF (7.0 mL) wereadded potassium phosphate (0.314 g, 1.48 mmol) , 2M solution of dimethylamine (0.5 mL , 0.990 mmol) andthe reaction mixture was heated at 80 °C for 12 h. The reaction mixture was quenched with ice and extracted with EtO Ac. Organic layer was washed with water, saturatedNaHC0 ₃ solution, brine, dried over sodium sulphate, and concentrated. The crude productwas purified by biotage purifier with 7% methanol in dichloromethane as eluent to afford the title compound as a viscous liquid (0.1 g, 67%): ¾ NMR (DMSO-d _6, 400 MHz) δ 1.79 (t, J = 7.2 Hz, 2H), 2.08 (s, 6H), 2.31 (t, J = 6.4 Hz, 2H), 3.98 (t, J = 6.8 Hz, 2H), 6.99 (t, J = 8.0 Hz, 1H), 7.09 (d, J = 8.0 Hz, 1H), 7.15-7.24 (m, 3H), 7.59 (d, J = 1.6 Hz, 1H), 7.71 (dd, J = 8.4 Hz, 1.6 Hz, 1H), 9.78 (s, 1H); MS (ESI) m/z 313 (M+H) ⁺ Step 4:3-(3-(Benzo[i/]thiazol-2-yl)-10H-phenothiazin-10-yl)-iV^V- dimethylpropan-l-amine

A solution of 10-(3-(dimethylamino)propyl)-10H-phenothiazine-3-carbaldehyd e (0.05 g, 0.16 mmol) and 2-amino-benzenethiol (0.024 g, 0.192 mmol) in DMSO (5.0 rriL) was heated at 150 °C for 12 h. The reaction mixture is quenched with ice, and extractedwith EtO Ac. Organic layer was washed with water, saturatedNaHCC solution, brine, dried over sodium sulphate, and concentrated. The crude productwas purified by biotage purifier with 7% methanol in dichloromethane as eluent to afford the title compound as a green solid (0.012 g, 18%): ¾ NMR (DMSO-d _6, 400 MHz) δ 1.78-1.85 (m, 2H), 2.08 (s, 6H), 2.32 (t, J = 7.2 Hz, 2H), 3.97 (t, J = 6.8 Hz, 2H), 6.98 (t, J= 7.6 Hz, 1H), 7.08 (d, J = 8.0 Hz, 1H), 7.16-7.18 (m, 2H), 7.23 (t, J= 7.6 Hz, 1H), 7.42 (t, J = 7.6 Hz, 1H), 7.51 (d, J = 7.2 Hz, 1H), 7.79 (d, J = 2.4 Hz, 1H), 7.87-7.89 (m, 1H), 7.99 (d, J= 8.0 Hz, 1H), 8.09 (d, J= 8.0 Hz, 2H); MS (ESI) m/z 418 (M+H) ⁺ ; HPLC purity: 99.38%.

Some examples of compounds synthesised by the method of Scheme VII are provided in Table VII.

Figure 8 shows general synthetic scheme VIII for the synthesis of selected 1,3,7-trisubstituted phenothiazenes.Neucleophilic substitution of substituted arylamino thiol (Villa) with aryl halide (Vlllb) followed by insitu Smiles rearrangement gave trisubstituted phenothiazines (VIIIc). Suzuki coupling of Vlllcwith arylboronic acid followed by reduction of nitro group with Pd/C givescompound Vllle. Reductive amination of Vlllewith cyclic ketones followed by deprotection using HC1 gave title compounds Vlllg.

Detailed synthetic descriptions of some compounds synthesised by the method of Scheme VIII are provided below.

Compound 212: 7-(lH-indol-2-yl)-iV-(piperidin-4-yl)-3-(trifluoromethyl)-10 H- phenothiazin-l-amine trifluoroacetic acid salt

Step 1: 7-Bromo-l-nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-amino-5-bromobenzenethiol (0.9 g, 4.41 1 mmol) in ethanol (30 mL) was added 2-chloro-l,3-dinitro-5-(trifluoromethyl)benzene (1.07 g, 3.369 mmol) and stirred for 20 min. To the reaction mixturewas added sodium hydroxide (0.52 g, 13.233 mmol) at 0 °C and was stirred for 16 h. The reaction mixture was filtered and dried under vacuum to give the title compound as a black colour solid (0.9 g, 52%): 1H NMR (DMSO-d _6, 400 MHz) δ 7.05 (d, J=8.4 Hz, 1H), 7.27-7.24 (m, 1H), 7.30 (d, J=2 Hz, 1H), 7.67 (s, 1H), 8.0 (s, 1H); MS (ESI) m/z 389.0 (M-H) ⁺

Step 2 : 7-(lH-Indol-2-yl)- l-nitro-3-(trifluoromethyl)- 1 OH-phenothiazine

To a stirred solution of 7-bromo-l-nitro-3-(trifluoromethyl)-l OH-phenothiazine (0.7 g, 1.789 mmol)) in 1 ,4-dioxane/water (2:1) wereadded (l-(/er/-butoxycarbonyl)-lH-indol-2-yl)boronic acid (0.7 g, 2.684 mmol) and potassium carbonate (0.74 g, 5.367 mmol) and the reaction mixture was purged with argon gas for 15 min. To the reaction mixturePd(dppf)Cl ₂ .DCM (0.073 g, 0.0894 mmol) was added and was stirred at 100 °C for 12 h in seal tube. The reaction mixture was washed with water, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S0 ₄ , concentrated, and the residue was purified by column chromatography over silica gel using ethyl acetate/hexane (1 :2.3) mixture as eluent to give the title compound as a light brown solid (0.15 g, 20%): MS (ESI) m/z 428.1 (M+H) ⁺ .

Step 3: 7-(l//-Indol-2-yl)-3-(trifluoromethyl)-10//-phenothiazin-l-a mine

To a stirred solution of 7-(lH-indol-2-yl)-l -nitro-3-(trifluoromethyl)-10H-phenothiazine (0.15 g, 0.351 mmol) in methanol (10 mL) was added 10% Pd/C and stirred for 16 h under hydrogen atmosphere at room temperature. The reaction mixture was filtered,filtrate was concentrated, and the residue was purified by column chromatography over silica gel using ethyl acetate/hexane (1 :2.3) mixture as eluent to give the title compound as a light green solid (0.01 g, 7%): H NMR (DMSO-d _6, 400 MHz) δ 5.43 (s, 2H), 6.52 (s, 1H), 6.72 (d, J=5.2 Hz, 2H), 6.88 (d, J=8.4 Hz, 1H), 6.94 (t, J=7.6 Hz, 1H), 7.02 (t, J=7.6 Hz, 1H), 7.31 (d, J=8.0 Hz, 1H), 7.43 (d, J=4 H, 1H), 7.45 (s, 1H), 7.49 (d, J=8.4 Hz, 1H), 7.92 (s, 1H), 11.3 (s, 1H);MS (ESI) m/z 398.3 (M+H) ⁺ ; HPLC purity: 95.09%.

Step 4: Tert-but i 2-(9-((l-(tert-butoxycarbonyl)piperidin-4-yl)amino)-7- (trifluoromethyl)10H-phenothiazin-3-yl)-lH-indole-l-carboxyl ate

To a stirred solution of 7-(lH-indol-2-yl)-3-(trifluoromethyl)-10H-phenothiazin-l-ami ne( 0.11 g, 0.22 mmol)in MeOH (10 mL) were added /er/-butyl 4-oxopiperidine-l-carboxylate(8.4 g, 42.5 mmol), AcOH (0.1 mL), and the reaction mixture was stirred for 2 h at 70 °C. The reaction mixture is cooled to room temperature, NaCNB¾ (0.069 g, 1.105 mmol) was added and stirring continued at 70 °C for 12 h. The reaction mixture was evaporated, diluted with ag.NaHCC solution, and extracted with DCM. The organic layer was dried over Na ₂ S0 ₄ , concentrated, and the residue was purified on silica gel using 10% ethyl acetate in hexane eluant to afford titlecompound(0.02 g, 16%):MS (ESI) «¾/z681 (M+H) ⁺ .

Step 5:7-(lH-Indol-2-yl)-/V-(piperidin-4-yl)-3-(trifluoromethyl)- 10H-phenothiazin-l-amine trifluoroacetic acid salt

To a stirred solution of tert-butyl 2-(9-((l-(tert-butoxycarbonyl)piperidin-4-yl)amino)-7- (trifluoromethyl)-10H-phenothiazin-3-yl)-lH-indole-l-carboxy late (0.02 g, 0.029 mmol) in dichloromethane (5 mL) at 0 °C was added trifluoroacetic acid (0.5 mL) and stirred for 1 h. The reaction mixture was concentrated, neutralized with sodium bicarbonate solution, extracted with 10% methanol in dichloromethane, the combined organic phase was dried over Na ₂ S0 ₄ , concentrated, and the residue was purified by preparative HPLC using 0.01% TFA in water/acetonitrile as eluent to give the title compound as a light black solid (0.004 g, 23%): 1H NMR (DMSO-d _6, 400 MHz) δ 5.43 (s, 2H), 6.52 (s, IH), 6.72 (d, J=5.2 Hz, 2H), 6.88 (d, J=8.4 Hz, IH), 6.94 (t, J=7.6 Hz, IH), 7.02 (t, J=7.6 Hz, IH), 7.31 (d, J=8.0 Hz, IH), 7.43 (d, J=4 Hz, IH), 7.45 (s, IH), 7.49 (d, J=8.4 Hz, IH), 7.92 (s, IH), 11.3 (s, IH): MS (ESI) m/z 481 (M, free base + H) ⁺ ; HPLC purity: 89. 9%.

Compound 242: iV-(7-(lH-Indol-2-yl)-3-(trifluoromethyl)-10H-phenothiazin-l -yl)-3- aminocyclohexanecarboxamide

Step 1: 7-Bromo-l-nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-amino-5-bromobenzenethiol(5 g, 24.50 mmol)) in ethanol (100 mL) was added 2-chloro-l,3-dinitro-5-(trifluoromethyl)benzene (5.95 g, 22.058 mmol) and stirred for 20 min. To the reaction mixture was added sodium hydroxide (2.94 g, 73.527 mmol) at 0 °C and stirred for 16 h at room temperature. The reaction mixture was filtered and dried under vacuum to give the title compound as a black color solid (5 g, 52%): ¾ NMR (DMSO-d ₆ , 400 MHz) δ 9.83 (s, 1H), 8.01 (s, 1H), 7.67 (s, 1H), 7.30 (d, J=2.4 Hz, 1H), 7.27-7.24 (m, 1H), 7.05(d, J=8.4 Hz, 1H); MS (ESI) m/z 388.9 (M-H) ⁺

Step 2 : 7-(lH-Indol-2-yl)- l-nitro-3-(trifluoromethyl)- 1 OH-phenothiazine

To a stirred solution of 7-bromo-l-nitro-3-(trifluoromethyl)-l OH-phenothiazine (1 g, 2.55 mmol)) in 1 ,4-dioxane/water (2:1)(15 mL) were added (l -(tert-butoxycarbonyl)-lH-indol-2- yl)boronic acid (1 g, 3.86 mmol) and potassium carbonate (1.05 g, 7.66 mmol)and the reaction mixture was purged with argon gas for 15 min. To the reaction mixture was addedPd(dppf)Cl ₂ .CH ₂ Cl ₂ (0.1 g, 0.12 mmol) and stirring continued at 100 °C for 12 h in a seal tube. The reaction mixture was washed with water, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S0 ₄ , concentrated, and the residue was purified by column chromatography over silica gel using ethyl acetate/hexane mixture as eluent to give the title compound as a block color solid (0.25 g, 23%):MS (ESI) m/z 426.0 (M-H) ⁺ .

Step 3: 7-(l//-Indol-2-yl)-3-(trifluoromethyl)-10//-phenothiazin-l-a mine

To a stirred solution of 7-(lH-indol-2-yl)-l -nitro-3-(trifluoromethyl)-10H-phenothiazine (0.25 g, 0.351 mmol) in methanol (10 mL) was added 10% Pd/C and stirred for 3 h under hydrogen atmosphere at room temperature. The reaction mixture was filtered, concentrated, and the residue was purified by column chromatography over silica gel using ethyl acetate/hexane (1 :2.3) mixture as eluent to give the title compound as a light green solid (0.07 g, 30%):Ή NMR (DMSO-d _6, 400 MHz) δ 5.43 (s, 2H), 6.52 (s, 1H), 6.72 (d, J=4.8 Hz, 2H), 6.88 (d, J=8.4 Hz, 1H), 6.94 (t, J=7.2 Hz, 1H), 7.02 (t, J=7.2 Hz, 1H), 7.31 (d, J=8.0 Hz, 1H), 7.48-7.42 (m, 2H), 7.50 (d, J=1.5 Hz, 1H), 7.92 (s, 1H), 11.3 (s, 1H);MS (ESI) m/z 398.0(M+H) ⁺

Step 4: Terf-butyl (3-((7-(lH-indol-2-yl)-3-(trifluoromethyl)-10H-phenothiazin- l- yl)carbamoyl)cyclohexyl)carbamate

To a stirred solution of 7-(lH-indol-2-yl)-3-(trifluoromethyl)-10H-phenothiazin-l-ami ne(0.07 g, 0.176 mmol)) in pyridine (3 mL) were added POCI ₃ (0.5 mL) and3-((tert- butoxycarbonyl)amino)cyclohexanecarboxylic acid (0.064 g, 0.264 mmol) and stirring continued for 2 h at room temperature. The reaction mixture was washed with water, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S04, concentrated, and the residue was purified by column chromatography over silica gel using ethyl acetate/hexane mixture as eluent to give the title compoundas a light brown solid (0.02 g, 22%):MS (ESI) m/z 624 (M+H) ⁺

Step 5: /V-(7-(lH-Indol-2-yl)-3-(trifluoromethyl)-10H-phenothiazin-l -yl)-3- aminocyclohexanecarboxamide:

To a stirred solution of tert-butyl (3-((7-(lH-indol-2-yl)-3-(trifluoromethyl)-10H-phenothiazin- l- yl)carbamoyl)cyclohexyl)carbamate(0.02 g, 0.0321 mmol)) in DCM (5 mL) was added triflouro acetic acid ( 0.5 ml) at 0 °C and stirring continued for 2 h at room temperature. The reaction mixture concentrated,basified with sodium bicarbonate solution, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S04, concentrated, and the residue was recrystalised by using w-pentane- diethylether to give the title compound as a light green solid (0.001 g, 63%): ¾ NMR (DMSO-d _6, 400 MHz) δ 1.21 (s, 2H), 1.75 (s, 4H), 1.87 (s, 2H), 1.98 (s, 2H), 6.75 (s, 1H), 6.94 (t, J=7.2 Hz, 1H), 6.95 (d, J=7.6 Hz, 2H), 7.05-7.02 (m, 1H), 7.48-7.42 (m, 2H), 7.16 (s, 1H), 7.32 (d, J=8 Hz, 1H), 7.36 (s, 1H), 7.45 (d, J=8 Hz, 2H), 7.52 (d, J=8.4 Hz, 1H), 8.09 (s, 1H), 11.33 (s, 1H);MS (ESI) m/z 523.2 (M+H) ⁺ ; HPLC Purity; 98.42 %.

Some examples of compounds synthesised by the method of Scheme VIII are provided in Table VIII.

Table VIII

Figure 9 shows general synthetic scheme IX for the synthesis of selected 1,3,8-trisubstituted phenothiazenes.Nucleophilic substitution reaction of substituted 2-amino thiophenol (IXa) with aryl halides (IXb) followed by insituSmiles rearrangement gave title compounds IXc.

Figure 10 shows general synthetic scheme X for the synthesis of selected 1,3,7-trisubstituted phenothiazenes.Nucleophilic substitution reaction of substituted 2-amino thiophenol (Xa) with aryl halides (Xb) followed by in szYwSmiles rearrangement gavetitle compounds Xc. Reduction of compound Xc using Pd/C followed by salt preparation resulted title compound Xe. Figure 11 shows general synthetic scheme XI for the synthesis of a selected 1, 3, 8-trisubstituted phenothiazineN-Acylation of 2-bromo-5-fluoroaniline (XIa) followed by neucleophilic substitution reaction with a thiol surrogate yielded compound Xld. Deprotection of alkyl chain using NaOEt followed by nucleophilic substitution and Smiles rearrangement give compound Xlg. Deprotection of compound Xlg using SOCbfollowed by reduction using Pd/C resulted compound Xli. Acid-amine coupling of Xli with Xlj followed by deprotection using HC1 yielded the title compound XII.

A detailed synthetic description of a compound synthesised by the method of Scheme XI is provided below.

Compound 124: 3-Amino-iV-(8-fluoro-3-(trifluoromethyl)-10H-phenothiazin-l- yl)cyclohexane carboxamide. TFA salt

TFA salt

Step 1: iV-(2-bromo-5-fluorophenyl) acetamide

To a stirred solution of 2-bromo-5-fluoroaniline (3.0 g, 15.95 mmol) in dichloromethane (50 mL) was added di isopropyl ethylamine(5.5mL,31.9mmol)) followed by acetyl chloride (1.7mL, 23.9 mmol) at 0 °C and starred at room temperature for 16h. The reaction mixture was washed with saturated sodium bicarbonate solution (50 mL), extracted the compound with dichloromethane, dried over sodium sulphate, filtered and concentrated. The crude product was purified on biotage with 6% ethyl acetate/n- hexane as eluant to get desired product 142a as off white solid. (3.0 g, 96%) . MS (ESI) m/z 233 (M+H) ⁺ .

Step 2: 3-Ethylheptyl 3-((2-acetamido-4-fluorophenyl)thio) propanoate

To a stirred solution of N-(2-bromo-5-fluorophenyl) acetamide ( 2.0 g, 0.086 mol) and 2- ethylhexyl 3-mercaptopropanoate in toluene (20 mL) was added DIPEA (4.6 mL 0.258 mol,) followed by Xanthpos (0.04 g, 0.086mmol) at room temperature. Then purged with N ₂ for 10 min, then added Pd ₂ (dba) 3 (0.078 g, 0.86 mol), then purged with N ₂ for 10 min. Reaction mixture was stirred at 110 °C for 4 h. The reaction mixture was filtered through celite, concentrated under reduced pressure. Obtained crude product was purified on biotage with 20% Ethyl acetate/hexane as eluant to give the desired productas as off white solid(.1.7 g, 55% ): ¹ H NMR (CDC1 _3, 400 MHz) δ 0.86-0.96.05 (m, 7H), 1.26-1.36 (m, 6H), 1.38-1.39 (m, 2H),1.54 (s, 1H), 2.25 (s,3H), 2.53 (t, J = 13.2 Hz, 2H), 2.93 (t, J = 6.8 Hz, 2H), 4.03-4.04 (m, 2H), 6.72 - 6.77(m, 1H), 7.22-7.27 (m, 1H), 7.50 (t, J = 6.4 Hz, 1H), 8.32 (t, J = 11.1 Hz, 1H), 8.77 (s, 1H).MS (ESI) m/z 370 (M+H) ⁺ .

Step 3: Sodium 2-acetamido-4-fluorobenzenethiolate

To a stirred sol of 3-ethylheptyl 3-((2-acetamido-4-fluorophenyl)thio) propanoate (0.7 g, 0.00189 mol) in ethanol (10 mL) was added sodium ethoxide solution in ethanol (lmL) at 0 °C, then stirred at 0 °C for 2 h, evaporated the solvent under reduced pressure. The crude (1.0 g) obtained was forwarded to the next step.

Step 4: l-(8-Fluoro-l-nitro-3-(trifluoromethyl)-10H-phenothiazin-10- yl) ethanone

A stirred suspension of sodium 2-acetamido-4-fluorobenzenethiolate (1.0 g, 1.89mmol) in DMF (5 mL) was heated at 100 °C for 14 h. To this added 50 mL of ice cold water, extracted the compound into EtOAc, dried the organic layer over sodium sulphate, filtered and concentrated. Obtained crude was purified on biotage with 30% EtOAc/hexane as eluant to give the product( 0.8 g, 80 %) , MS (ESI), m/z 373 (M+H) ⁺ .

Step 5: 8-Fluoro-l-nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of l -(8-fluoro-l-nitro-3-(trifluoromethyl)-10H-phenothiazin-10-y l) ethanone (0.8 g, 0.00215 mol) in methanol (30 mL) was added thionyl chloride (6 mL) at 0 °C and starred at 70 ° for 18 h. The reaction mixture was concentrated under reduced pressure, quenched the reaction mixture with bicarbonate solution extracted the compound into EtOAc, dried the organic layer over sodium sulphate, filtered and concentrated. Obtained crude was purified on biotage with 20% EtOAc/hexane as eluant to to give the product( 0.3 g, 42 %), MS (ESI), m/z 329 (M-H) ⁺ .

Step 6: 8-Fluoro-3-(trifluoromethyl)-10H-phenothiazin-l-amine

To a stirred solution of 8-fluoro-l -nitro-3-(trifluoromethyl)-10H-phenothiazine(142e, 0.3 g, 0.90 mmol) in ethyl acetate (20 mL) was added a solution of pd/C (50%, water wet, 0.2 g) , and stirred at room temperature for 4 h. The reaction mixture was filtered through celite and concentrated to offer the title compound. (0.3 g crude). MS (ESI), m/z 301 (M+H) ⁺ . Step 7: 7eri-butyl(3-((8-fluoro-3-(trifluoromethyl)-10H-phenothiazin -l- yl)carbamoyl)cyclohexyl) carbamate

To a stirred solution of 8-fluoro-3-(trifluoromethyl)-10H-phenothiazin-l -amine(142f, 0.2g, 0.6 mmol) and 3-((tert-butoxycarbonyl)amino)cyclohexanecarboxylic acid(0.2 g, 0.82 mmol) in pyridine(3 mL) was added a solution POCI ₃ (0.3 mL) at 0 °C and starred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure, added 20 mL of DM water , extracted compound with EtOAc, dried the organic layer over sodium sulphate filtered and concentrated. The crude was purified on biotage with 30 %EtOAc/hexane as eluant to get title compound 142 as off white solid (0.060 g, 17%). MS (ESI), m/z 524 (M-H) ⁺ .

Step 8: 3-Amino-N-(8-fluoro-3-(trifluoromethyl)-10H-phenothiazin-l- yl)cyclohexanecarboxamide. TFA salt

TFA salt

To a stirred solution of tert-butyl(3-((8-fluoro-3-(trifluoromethyl)-10H-phenothiazin -l- yl)carbamoyl)cyclohexyl)carbamate (0.060 g, 0.11 mmol) in dichloromethane (6 mL) was added a solution of HCl/dioxane (20%, 0.5 mL) at 0 °C and starred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure, dissolved in saturated sodium bicarbonate solution, extracted with dichloromethane, and concentrated. The crude was purified by prep TLC to get the title compound (0.020 g, 41%). ¾ NMR (DMSO-i _6, 400 MHz) δ 1.36- 1.49 (m ,3 H), 1.80-2.01 (m, 5H),2.11-2.31 (m, 1H), 3.089 (bs, 1H), 6.70-6.77 (m, 2H), 7.00- 7.04 (m, 1H), 7.20(s, 1H), 7.30(s, 1H), 7.77 (bs, 3H), 8.18(s, 1H), 9.04 (s, 1H): MS (ESI) m/z 426.1(M+H) ^" ; HPLC purity: 99.1%. Figure 12 shows general synthetic scheme XII for the synthesis of selected triazolo phenothiazenes and imidazolo phenothiazenes.Nucleophilic substitutionof 2-amino thiophenol (Xlla) with aryl halides (Xllb) followed by in szYwSmiles rearrangement give compound XIIc. Reduction of compound XIIc using Pd/C resulted compound Xlld. Cyclization of compound Xlld with aldehydes (B) followed by deprotection resulted imidazolo phenothiazenes (XHg). Parallelly, cyclization of compound Xlld with nitroso compound (C) yilded triazolo phenothiazene (Xllf).

A detailed synthetic description of a compound synthesised by the method of Scheme ΧΠ is provided below. Compound 239: 2-(4-(4-(Trifluoromethyl)imidazo[4,5,l-^/]phenothiazin-l-yl) piperidin-l- yl)ethanamine

Step l:l-Nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-chloro-l,3-dinitro-5-(trifluoromethyl)benzene (20 g, 74.07 mmol) in water (200 mL) were added 2-aminobenzenethiol (7.9 mL, 74.07 mmol), sodium hydroxide (8.8 g, 222 mmol) and the reaction mixture was heated at 100 °C for 16 h. The reaction mixture was cooled to room temperatur, the residue was filtered, and washed with EtOH followed by H ₂ 0 to give the title compound as a brown solid (20 g, 86%). The product spot was matched with auenthetic in TLC.

Step 2: 3-(Trifluoromethyl)-10H-phenothiazin-l -amine

To a stirred solution of l -nitro-3-(trifluoromethyl)-10H-phenothiazine(25 g, 18.128 mmol) in MeOH (300 mL) was added 10% Pd/C (50% wet, 5.0 g) and the reaction mixture was stirred at room temperature for 24 h under ¾ atmosphere. The reaction mixture was filtered through celite and filtrate was concentrated to give the title compound as a light brown solid (15 g, 67%): 1H NMR (DMSO-d ₆ , 400 MHz) δ 5.42 (s, 2H), 6.48 (s, 1H), 6.72 (s, 1H), 6.77 (t, J = 7.2 Hz, 1H), 6.83 (d, J=8 Hz, 1H), 6.92 (d, J = 8 Hz, 1H), 7.02 (t, J = 7.6 Hz, 1H), 7.79 (s, 1H); MS (ESI) m/z 283 (M+H) ⁺ .

Step 3: Terf-butyl 4-(4-(trifluoromethyl)imidazo[4,5,l-^/]phenothiazin-l-yl)pip eridine-l- carboxylate

Boc

To a stirred solution of 3-(trifluoromethyl)-10H-phenothiazin-l-amine(0.8 g, 2.836 mmol)) in ethanol (30 mL) and pyrrolidine (0.23 mL, 2.836 mmol) was added feri-butyl 4- formylpiperi dine- 1-carboxy late (0.54 mL, 2.836 mmol) and stirring continued for 12 h at 70 °C. The reaction mixture was concentrated, diluted with water,extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S0 ₄ , concentrated, and the residue was purified by column chromatography over silica gel using 10% ethyl acetate/hexane mixture as eluent to give the crude compound as a light brown liquid (0.3 g, crude): MS (ESI) m/z 476.1 (M+H) ⁺ Step 4: l-(Piperidin-4-yl)-4-(trifluoromethyl)imidazo[4,5,l-^/]pheno thiazine

To a stirred solution of tert-butyl 4-(4-(trifluoromethyl)imidazo[4,5,l-A:/]phenothiazin-l- yl)piperidine-l-carboxylate(0.3 g, crude) in dichloromethane (15 mL) at 0 °C was added 4N HC1 in 1,4-dioxane (1 mL) and stirring continued for 16 h at room temperature. The reaction mixture was concentrated washed with sodium bicarbonate solution, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ SC>4, concentrated, and the residue residue was purified by column chromatography over silica gel using 10%MeOH-DCMsolvent system as eluent to give the compound as a off white solid (0.035 g, 15%): ^X H NMR (DMSO-d _6, 400 MHz) δ 1.76 (t, J=10.8 Hz, 2H), 1.99 (d, J=12.8 Hz, 2H), 2.71 (t, j=12 Hz, 2H), 3.02 (d, j=11.6 Hz, 2H), 3.51 (s, 1H), 7.17 (t, J=7.6 Hz, 1H), 7.23 (s, 1H), 7.31 (s, 2H), 7.56 (s, 1H), 7.66 (s, J=8.4 Hz, 1H);MS (ESI) m/z 376.1 (M+H) ⁺ .

Step 5: Jer^butyl(2-(4-(4-(trifluoromethyl)imidazo[4,5 -^/]phenothiazin-l-yl)piperidin-l- yl)ethyl) carbamate

To a stirred solution ofl-(piperidin-4-yl)-4-(trifluoromethyl)imidazo[4,5,l-A:/]ph enothiazine (0.15 g, 0.4 mmol)) in acetonitrile (10 mL) were added potassium carbonate (0.16 g, 1.2 mmol),feri-butyl (2-bromoethyl)carbamate (0.13 g, 0.6 mmol) and stirring continued for 16 h at 80 °C. The reaction mixture was diluted with water, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S0 ₄ , concentrated, and the residue was purified by column chromatography over silica gel using 100% ethyl acetate mixture as eluent to give the title compound as a off white solid (0.17 g, 85%): ¾ NMR (CDC1 _3, 400 MHz) δ 1.46 (s, 9H), 2.17 (d, J=6.8 Hz, 4H), 2.4 (m, 1H), 2.53 (s, 2H), 3.07 (s, 2H), 3.25 (s, 4H), 6.94 (s, 1H), 7.23 (s, 1H), 7.09 (t, J=7.6 Hz, 1H), 7.17 (t, J=6.8 Hz, 2H), 7.45 (d, J=8 Hz, 2H), 7.54 (s, 1H); MS (ESI) m/z 519.1 (M+H) ⁺

Step 6: 2-(4-(4-(Trifluoromethyl)imidazo[4,5,l-^/]phenothiazin-l-yl) piperidin-l- yl)ethanamine

To a stirred solution of tert-butyl (2-(4-(4-(trifluoromethyl)imidazo[4,5,l-A:/]phenothiazin-l- yl)piperidin-l -yl)ethyl)carbamate (0.15 g, 0.289 mmol) in dichloromethane (10 mL) at 0 °C was added TFA (1 mL) and stirring continued for 5 h at room temperature. The reaction mixture was concentrated, washed with sodium bicarbonate solution, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S0 ₄ , concentrated, and the residue was recrystalised by using diethyl ether and pentane to give the title compound as a off white solid (0.03 g, 25%): ¹ H NMR (DMSO-d _6, 400 MHz) δ 1.91-1.83 (m, 2H), 2.05 (d, J=11.6 Hz, 2H), 2.16 (t, J=11.2 Hz, 2H), 2.35-2.30 (m, 2H), 3.51 (s, 1H), 2.62 (t, J=6.8Hz, 2H), 2.92 (d, J=11.2 Hz, 2H), 3.39 (d, j=11.6 Hz, 1H), 7.17 (t, j=7.2 Hz, 1H), 7.23 (s, 1H), 7.31 (t, j=7.6 Hz, 2H), 7.56 (s, 1H), 7.64 (d, J=8.4 Hz, 1H);MS (ESI) m/z 419.2 (M+H) ⁺ ; HPLC purity: 99.48%.

Some examples of compounds synthesised by the method of Scheme ΧΠ are provided in Table XII.

Table XII

Figure 13 shows general reaction scheme XIII for the synthesis of a selected triubstituted phenothiazine. Nucleophilic substitution reaction of substitutedamino thiols Xllla with substituted dinitro aryl halidesXIIIb, followed bu insituSmiles rearrangement resulted in the formation of trisubstituted phenothiazenes XIIIc. Nitro group reduction withPd/C gave compoundXIIId, which on reductive amination with an appropriate ketone resulted in corresponding amine compound (XHIe), followed by deprotection resultedin title compound xinf

A detailed synthetic description for a compound synthesised by the method of Scheme ΧΠΙ is provided below.

Compound 226: iV-(l-(2-Aminoethyl)piperidin-4-yl)-7-chloro-3-(trifluoromet hyl)-10H- phenothiazin-l-amine

Step 1: 7-Chloro-l-nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-amino-5-chlorobenzenethiol (5 g, 31.32 mmol) in EtOH (50 mL) were added 2-chloro-l ,3-dinitro-5-(trifluoromethyl)benzene (8.4 g, 31.32 mmol), NaOH (2.5 g, 62.64 mmol) and the reaction mixture was heated at 85 °C for 12 h. The reaction mixture was cooled to room temperature, concentrated,and the residue was purified by column chromatography over silica gel using dichloromethane/hexane (2%) mixture as eluent to give the title compound234»as a black solid (3 g, 46%): ¾ NMR (DMSO-d _6, 400 MHz) δ 7.10-7.14 (m, 2H), 7.18 (s, 1H), 7.67 (s, 1H), 8.00 (s, 1H), 8.71 (bs, 1H);MS (ESI) m/z 345 (M-H) ⁺ .

Step 2: 7-Chloro-3-(trifluoromethyl)-10H-phenothiazin-

To a stirred solution of 7-chloro-l -nitro-3-(trifluoromethyl)-10H-phenothiazine (1 g, 2.88 mmol) in MeOH (50 mL) were added Zn powder (1 g, 15.78 mmol),ammoniumchloride (0.2 g, 4.154 mmol) and stirring continued at room temperature for 12 h. The reaction mixture was filtered through celite, concentrated to give the title compound as a black solid (1 g, crude): ¹ H NMR (DMSO-d ₆ ,400 MHz) δ 5.41 (bs, 2H), 6.49 (s, 1H), 6.73 (s, 1H), 6.81 (d, J=8.4 Hz, 1H), 7.02- 7.06 (m, 2H), 7.91 (bs, 1H);MS (ESI) m/z 316 (M+H) ⁺ .

Step 3: Tert-butyl (2-(4-((7-chloro-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)piperidin-l-yl)ethyl)carbamate

To a stirred solution of 7-chloro-3-(trifluoromethyl)-10H-phenothiazin-l -amine (0.5 g, 1.57 mmol) and tert-butyl (2-(4-oxopiperidin-l-yl)ethyl)carbamate (0.75 g, 3.14 mmol) in methanol (50 mL) was added acetic acid (0.47 mL) and stirring continued for 1 h at room temperature. To the reaction mixture was added NaCNB¾ (0.49 g, 7.89 mmol) and was heated to 80°C for 12 h. The reaction mixture was concentrated and washed with sodium bicarbonate solution, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S04, concentrated, and theresidue was purified by column chromatography over silica gel using MeOH/DCM (3%) mixture as eluent to give the title compound as a black solid (0.1 g, 11%):MS (ESI) m/z 545 (M- H) ⁺ .

Step 4: iV-(l-(2-Aminoethyl)piperidin-4-yl)-7-chloro-3-(trifluoromet hyl)-10H-phenothiazin- 1-amine

To a stirred solution of (2-(4-((7-chloro-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)piperidin-l-yl)ethyl)carbamate (0.1 g, 0.18 mmol) in dichloromethane (10 mL) at 0 °C was added 4N HC1 in dioxane (0.5 mL) and stirring continued for 4 h at room temperature. The reaction mixture was nutralised with sodium bicarbonate solution, solid was filtered, and dried under vaccumto gave title compound as a off white solid (0.006 g, 8%):NMR (DMSO-d _6, 400 MHz) δ 1.39-1.47 (m, 2H), 1.88-1.91 (m, 2H), 1.96-1.98 (m, 2H), 2.06-2.09 (m, 2H), 2.23-2.27 (m, 2H), 2.61 -2.65 (m, 2H), 2.81-2.84 (m, 2H), 6.56 (d, J=15.2 Hz, 2H), 6.86 (d, J= 8.4 Hz, 1H), 7.04 (s, 1H), 8.11 (s, 1H);MS (ESI) m/z 443.1 (M+H) ⁺ ; HPLC purity: 99.14%.

Figure 14 shows general synthetic scheme XIV for the synthesis of selected triubstituted phenothiazenes. Aqueous hydrolysis of compound XlVa, gave substituted amino thiols XIVb,which on nucleophilic substitution reactionwith substituted dinitro arylhalidesXIVc, followed bu insituSmiles rearrangement resulted in the formation of trisubstituted phemothiaxzenes XlVd. Nitro group reduction withPd/C gave compound XlVf, Reductive amination of XlVfwith an appropriate ketone resulted in compound XlVfg, followed by deprotection resulted compound XlVj. Alternatively, compound XlVfg was reduced by Rany- Ni to the corresponding amine, followed by deprotection gave the title compound XlVi. Detailed synthetic descriptions forsome compounds synthesised by the method of Scheme XIV are provided below.

Compound 227: iV-(l-(2-Aminoethyl)piperidin-4-yl)-7-(aminomethyl)-3-(trifl uoromethyl)- lOH-phenothiazin-l-amine bis(2,2,2-trifluoroacetate)

Step 1: 2-Amino-5-bromobenzenethiol To a stirred solution of 6-bromobenzo[i ]thiazol-2-amine (1 g, 4.364 mmol) in water (20 mL) was added KOH (7.34 g, 130.947 mmol) at 0 °C, reaction mixture was heated at 120 °C for 16 h . The reaction mixture was acidified with glacial acetic acid (up to pH~6) at 0 °C, solid was filterd, and dried under vacuum to give crude title compound as a light green solid (0.95 g, crude);MS (ESI) m/z 201 (M-H) ⁺

Step 2: 7-Bromo-l-nitro-3-(trifluoromethyl)-10H-phenothiazine

To a stirred solution of 2-amino-5-bromobenzenethiol(0.9 g, 4.411 mmol) in ethanol (30 mL) was added NaOH (0.52 g, 13.23 mmol) at 0 °C and stirring continued for 20 min at 0 °C. To the reaction mixturewas added 2-chloro-3-nitro-5-(trifluoromethyl)aniline (1.07 g, 3.969 mmol) and stirring continued at room temperature for 16 h. The reaction mixture was filtered and solid was washed with H ₂ 0 to gave the title compound as a brown solid (0.9 g, 52%): 1H NMR (DMSO-d _6, 400 MHz) δ 7.05 (d, J=7.8 Hz, 1H), 7.27-7.24 (m, 1H), 7.30 (d, J=2 Hz, 1H), 7.67 (s, 1H), 8.00 (s, 1H), 9.83 (s, 1H); MS (ESI) m/z 389.0 (M-H) ⁺ .

Step 3: 9-Nitro-7-(trifluoromethyl)-10H-phenothiazine-3-carbonitrile

To a stirred solution of 7-bromo-l-nitro-3-(trifluoromethyl)-10H-phenothiazine (0.8 g, 2.04 mmol)) in DMF (15 mL) was added CuCN (0.36 g, 4.09 mmol) and reaction mixture was heated at 150 °C for 16 h. The reaction mixture was washed with water, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S0 ₄ , concentrated, and the residue was purified by column chromatography over silica gel using ethyl acetate/hexane (1 :2.3) mixture as eluent, to give the title compound as a black solid (0.5 g, 72%): ¾ NMR (DMSO-d _6, 400 MHz) δ 7.08 (d, J= 6.4 Hz, 1H), 7.24 (d, J= 6.8 Hz, 1H), 7.30 (s, 1H), 7.67 (s, 1H), 8.01 (s, 1H), 9.83 (s, 1H).

Step 4: 9-Amino-7-(trifluoromethyl)-10H-phenothiazine-3-carbonitrile

To a stirred solution of 9-nitro-7-(trifluoromethyl)-10H-phenothiazine-3-carbonitrile (0.7 g, 2.07 mmol) in methanol (10 mL) were added zinc powder (0.13 g, 2.077 mmol), ammoniumchloride (0.2 g, 4.154 mmol) and stirring continued for 4 h at RT. The reaction mixture was filtered through celite, concentrated, and the residue was purified by column chromatography over silica gel using ethyl acetate/hexane (1 :2.3) mixture as eluent to give the title compound as a brown solid (0.4 g, 63%): MS (ESI) m/z 306.0 (M-H) ⁺ . Step 5: Tert-butyl (2-(4-((7-cyano-3-(trifluoromethyl)-10H-phenothiazin-l-yl)am ino) piperidin-l-yl)ethyl)carbamate

To a stirred solution of 9-amino-7-(trifluoromethyl)-10H-phenothiazine-3-carbonitrile (3, 0.2 g, 0.65 mmol)) in dichloroethene (3 mL) at 0 °C were added tert-butyl (2-(4-oxopiperidin-l - yl)ethyl)carbamate (4, 0.31 g, 1.30 mmol), acetic acid (0.3 mL) and stirring continued for 5 min. To the reaction mixture was added sodiumborohydride (0.05 g, 1.30 mmol) at 0 °C and stirring continued at room temperature for 2 h. The reaction mixture was washed with sodium bicarbonate solution, extracted with dichloromethane, the combined organic phase was dried over Na ₂ SC>4, concentrated, and the residue was purified by column chromatography over silica gel using methnoal/dichloromethane (10 %) mixture as eluent to give the title compound (0.07 g, 20%): ¾ NMR (DMSO-d _6, 400 MHz) δ 1.35 (s, 9H), 1.41 (s, 2H), 1.88 (bs, 4H), 2.07 (bs, 2H), 2.39 (m, 2H), 2.47 (bs, 2H), 2.81 (bs, 2H), 3.00 (bs, 1H), 5.25 (d, J = 6.8 Hz, 1H), 6.54 (s, 1H), 6.59 (s, 1H), 6.90 (d, J = 8.4 Hz, 1H), 7.39-7.43 (m, 2H), 8.45 (s, 1H).

Step 6: Tert-buty\ (2-(4-((7-(aminomethyl)-3-(trifluoromethyl)-10H-phenothiazin -l-yl) amino) piperidin-l-yl)ethyl)carbamate

To a stirred solution of tert-bxxXy\ (2-(4-((7-cyano-3-(trifluoromethyl)-10H-phenothiazin-l - yl)amino)piperidin-l -yl)ethyl)carbamate (0.08 g, 0.15 mmol) in MeOH (15 mL) was added raney Ni (0.05 g) and hydrogenated under H ₂ atmosphere at 20 psi pressure& room temperature. The reaction mixture was filtered through celite and filtrate was evaporated to get title compoundas an light brown solid(0.08 g, crude): MS (ESI) m/z 538 (M+H) ⁺ .

Step 7: iV-(l-(2-Aminoethyl)piperidin-4-yl)-7-(aminomethyl)-3-(trifl uoromethyl)-10H-phen othiazin-l-amine

To a stirred solution of tert-butyl (2-(4-((7-(aminomethyl)-3-(trifluoromethyl)-10H- phenothiazin-l-yl)amino)piperidin-l-yl)ethyl)carbamate (0.08 g, 0.14 mmol) in dichloromethane (3 mL) at 0 °C was added HCl/dioxane (0.4 mL) and stirring continued for 4 h. The reaction mixture was evaporated, residue was washed with sodium bicarbonate solution, extracted with dichloromethane, the combined organic phase was dried over Na ₂ S0 ₄ , and concentrated to gave the crude compound. Crude compound was purified by prep-HPLC to give title compound as an brown solid (0.002 g, 4%): ¹ H NMR (DMSO-d _6, 400 MHz) δ 1.63-1.72 (m, 2H), 1.94-1.98 (m, 2H), 2.07-2.18 (m, 2H), 3.12-3.33 (m, 4H), 3.63-3.73 (m, 2H), 3.83-3.84 (m, 2H), 5.42-5.47 (m, 1H), 6.89-6.63 (m, 2H), 6.85-6.87 (m, 1H), 7.04-7.07 (m, 2H), 7.97 (bs, 4H, salt);MS (ESI) m/z 436.4 (M-H) ⁺ ; HPLC purity: 97.03%.

Compound217: 9-((l-(2-Aminoethyl)piperidin-4-yl)amino)-7-(trifluoromethyl )-10H- phenothiazine-3-carbonitrile

To a stirred solution of tert-butyl (3-(((3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)methyl)cyclohexyl)carbamate (186, 0.06 g) in dichloromethane (5 mL) at 0 °C was added HCl/dioxane solution(0.3 mL) and stirring continued for 4 h. The reaction mixture was evaporated, residue was washed with sodium bicarbonate solution, extracted with dichloromethane, the combined organic phase was dried over Na ₂ S04, concentrated, to give the title compound as a light brown solid (0.03 g, 63%): ¾ NMR (DMSO-d _6, 400 MHz) δ 1.42-1.45 (m, 2H), 1.88 (bs, 2H), 2.07 (d, J = 1 1.2 Hz, 2H), 2.30-2.33 (m, 2H), 2.65 (d ,J = 8.0 Hz, 2H), 2.81 (bs, 2H), 5.32 (d, J = 6.4 Hz, 1H), 6.54 (s, 1H), 6.59 (s, 1H), 6.93 (d, J = 8.4 Hz, 1H), 7.39- 7.43 (m, 2H), 8.51 (bs, 1H).MS (ESI) m/z 434.2 (M+H) ⁺ ; HPLC purity: 98.8%.

Figure 15 shows general synthetic scheme XV for the synthesis of selected trisubstituted phenothiazenes.Bromination of phenothiazene XVa with Br ₂ gave the dibromo phenothiazene XVb which on Suzuki coupling with indole boronic acid gave gompound XVc. N-10 alkylationof XVc with dihalide followed by azide formation gave compound XVe. Azide reduction followed by amine protection gave compound XVg. Palladium catalyzed amination/cayanation followed by deprotection gave the title compound XVi.

A detailed synthetic description of a compound synthesised by the method of Scheme XV is provided below.

Compound244: 3-(3-(lH-indol-2-yl)-7-(piperazin-l-yl)-10H-phenothiazin-10- yl)propan-l- amine

Step 1: 3,7-Dibromo-lOH-phenothiazine To a stirred solution of 1 OH-phenothiazine (5 g, 25.1 mmol)in AcOH (50 mL) was added Br ₂ (3.3 mL, 63 mmol) and stirring continued at room temperature for 16 h. The reaction mixture was filtered and dried to get the title compound asbrown solid(7 g, quantitative) ^HNMR (DMSO _, 400 MHz) δ 6.55-6.57 (m, 2H),7.06-7.20 (m, 4H), 8.79 (s, 1H);MS (ESI) «¾/z358 (M+2H) ⁺ .

Step 2: Tert-butyl 2-(7-bromo-10H-phenothiazin-3-yl)-lH-indole-l-carboxylate

To a stirred solution of 3,7-dibromo-10H-phenothiazine(3.5 g, 9.80 mmol) and (\-(tert- butoxycarbonyl)-lH-indol-2-yl)boronic acid (3.8 g, 14.7 mmol) in a mixture of 1,4- dioxane/water (55/5 mL) mixturewas added potassium carbonate (4 g, 29.4 mmol) and was purged with nitrogen for 15 mins. To the reaction mixture was added tetrakis(triphenylphosphine)palladium(0)(l . l g, 0.98 mmol), was purged with nitrogen for 10 mins, and heated at 100 °C for 12hin a seal tube. The reaction mixture was filtered through celite, filtrate was diluted with EtOAc, and washed with water followed by brine.The organic layer was dried over sodium sulphate and concentrated. The crude was purified over silica gel using 12% EtOAc/hexane as eluant to give title compound as brown solid(1.5 g, 31%): MS (ESI) m/z496 (M+2H) ⁺ .

Step 3: Tert-butyl 2-(7-bromo-10-(3-chloropropyl)-10H-phenothiazin-3-yl)-lH-ind ole-l- carboxylate

To a stirred solution of tert-butyl 2-(7-bromo-10H-phenothiazin-3-yl)-lH-indole-l -carboxylate (245bjl.5 g, 3.04 mmol) in DMF (30 mL) was added sodium hydride (0.18 g, 4.56 mmol) at 0 °C in small portions and stirring continued at 0 °C for 15 mins. To the reaction mixture was added 3-bromo chloropropane (0.57 g, 3.64mmol) at 0 °C and stirring continued at room temperature for 1 h. The reaction mixture was quenched with a</.NH ₄ Cl solution, extracted with EtOAc, the organic layer was washed with brine, dried over anhydrous sodiumsulphate, and concentrated to gave the title compound as an pale yellow solid (1.5 g, crude). The crude product was as such taken to the next step without further purification.MS (ESI) m/z370 (M+2H) ⁺ .

Step 4: Tert-butyl 2-(10-(3-azidopropyl)-7-bromo-10H-phenothiazin-3-yl)-lH-indo le-l- carboxylate

To a stirred solution of tert-butyl 2-(7-bromo-10-(3-chloropropyl)-10H-phenothiazin-3-yl)-lH- indole-l -carboxylate(24#e^l.5 g, 2.63 mmol) in DMSO (30 mL) was added sodium azide (0.51 g, 7.89 mmol) and the reaction mixture was heated at 80°C for 2 h. The reaction mixture was quenched with ice, extracted with EtOAc, the organic layer was dried over sodium sulphate, and concentrated. The crude product was purified over silica gel using 5% EtOAc/hexane as eluant to give the title compound as brown solid (0.9 g, 59%):MS (ESI) m/z576 (M+2H) ⁺ .

Step 5: Tert-butyl 2-(10-(3-aminopropyl)-7-bromo-10H-phenothiazin-3-yl)-lH-indo le-l- carboxylate

To a stirred solution ofiteri-butyl 2-(10-(3-azidopropyl)-7-bromo-10H-phenothiazin-3-yl)-lH- indole-l -carboxylate(34#d 0.9 g, 1.56 mmol) in a mixture of THF (20 mL) and H ₂ 0 (5 mL) was added triphenylphosphine (0.81 g, 3.12 mmol) and stirring continued at room temperature for 12 h. The reaction mixture was diluted with water, extracted with ethyl acetate, the organic layer is dried over sodium sulphate, and concentrated to get the title compound as an viscous solid (lg, crude). Compound was taken to next step without further purification.MS (ESI) m/z550 (M+2H) ⁺ .

Step 6: Tert-but i 2-(7-bromo-10-(3-((tert-butoxycarbonyl)amino)propyl)-10H- phenothiazin-3-yl)-lH-indole-l-carboxylate

To a stirred solution of tert-butyl 2-(10-(3-aminopropyl)-7-bromo-10H-phenothiazin-3-yl)-lH- indole-l -carboxylate(345e ₇ l g, 1.81 mmol) in dichloromethane (30 mL) were added Et ₃ N (0.5 mL, 3.63 mmol), (Boc) ₂ 0 (0.8 mL, 3.63 mmol) at 0 °C and stirring continued at room temperature for 2 h. The reaction mixture was concentrated andthe crude product was purified over silica gel using 15% EtOAc in hexane as eluant to get the title compound as yellow solid(0.65 g, 55%): ^]^ (DMSO. 400 MHz) δ 1.28 (s, 9H), 1.33 (s, 9H), 1.78-1.81 (m, 2H), 3.02-3.03 (m, 2H), 3.87-3.89 (m, 2H), 6.67 (s, 1H), 6.85 (br s, 1H), 6.95 (d, J=8.4 Hz, 1H), 7.06 (d, J=8.4 Hz, lH),7.13-7.34 (m, 6H),7.56 (d, J=7.6 Hz, 1H), 8.04 (d, J=8.4 Hz, 1H). Step 7: Tert-butyl 2-(10-(3-((tert-butoxycarbonyl)amino)propyl)-7-(4-(tert- butoxycarbonyl)piperazin-l-yl)-10H-phenothiazin-3-yl)-lH-ind ole-l-carboxylate

To a stirred solution of tert-butyl 2-(7-bromo-10-(3-((tert-butoxycarbonyl)amino)propyl)-10H- phenothiazin-3-yl)-lH-indole-l-carboxylate(0.23 g, 0.35 mmol) in a mixture of o-xylene (15mL) were added tert-butyl piperazine-l-carboxylate (0.16 g, 0.88 mmol), cesium carbonate (0.57 g, 1.76 mmol),BINAP (0.0065 g, 0.01 mmol) and reaction mixture was purged with nitrogen for 15 mins. To the reaction mixture was added tris(dibenzylideneacetone)dipalladium (0.1 g, 0.17 mmol), was purged with nitrogen for 10 mins, and heated at 100 °C for 12hin a seal tube. The reaction mixture was filtered through celite, filtrate was diluted with EtOAc, washed with water followed by brine. The organic layer was dried over sodium sulphate, and concentrated. The crude was purified over silica gel using 15% EtOAc/hexane as eluant to give title compound as brown sohd(0.15 g, 56%): MS (ESI) m/z756 (M+H) ⁺ .

Step 8 : 3-(3-(lH-Indol-2-yl)-7-(piperazin- 1-yl)- 1 OH-phenothiazin- 10-yl)propan- 1-amine

O

3

To a stirred solution of tert-butyl 2-(10-(3-((tert-butoxycarbonyl)amino)propyl)-7-(4-(tert- butoxycarbonyl)piperazin-l -yl)-l 0H-phenothiazin-3-yl)-lH-indole-l -carboxylate(0.15 g, 0.19 mmol) in dichloromethane (10 niL) was added TFA (2 niL) at 0 °C and stirring continued at room temperature for 12 h. The reaction mixture was concentrated, residue was basified with saturated sodium bicarbonate solution, extracted with dichloromethane, the organic layer was dried over sodium sulphate, and concentrated. Crude product was purified by prep. HPLC to give the title compound as brownsolid (25 g, 22%):1H NMR (DMSO-i/ ₆ , 400 MHz) δ 1.93-1.97 (m, 2H), 2.85-2.95 (m, 2H), 3.20-3.23 (m, 8H), 6.79 (s, 1H), 6.84-6.87 (m, 2H), 6.93-6.98 (m, 2H),7.02-7.09 (m, 2H), 7.33-7.47 (m, 2H), 7.65-7.68 (m, 2H),7.72 (bs, 3H, TFA salt), 8.78 (bs, 2H, TFA salt), 11.39 (s, 1H); MS (ESI) m/z 456.2 (M+H) ⁺ ; HPLC purity: 99.38%.

Some examples of compounds synthesised by the method of Scheme XV are provided in Table . Table XV

Figure 16 shows general synthetic scheme XVI for the synthesis of selected triubstituted phenothiazenes. Substituted benzothiazoles XVIa were hydrolysed with potassium hydroxide to the corresponding aminothiols XVIb, which on neucleophilic substitution with aryl halides followed by insituSmiles rearrangement gave the corresponding substituted phenothiazene XVIc. Esterification of XVIc followed by reduction of nitro group gave the corresponding aminesXVIe, which on reductive amination with appropriate carbohyl comounds gave the copound XVIf, which is deprotected with acid to give the title compound XVIg.

A detailed synthetic description of a compound synthesised by the method of Scheme XVI is provided below. Compound 247: Methyl 9-((l-(2-aminoethyl)piperidin-4-yl)amino)-7-(trifluoromethyl )- 10H-phenothiazine-3-carboxylate

Step 1: 4-Amino-3-mercaptobenzoic acid

A solution of methyl 2-aminobenzo[i/]thiazole-6-carboxylate (2 g, 9.61 mmol)) in water (25 mL) was heated at 100°C for 16 h. Reaction mixture was neutralized with citric acid, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S0 ₄ , and concentrated to get the title compoundas off white solid (1.625 g, 90%):MS (ESI) m/z 170 (M+H) ⁺ .

Step 2: 9-Nitro-7-(trifluoromethyl)-10H-phenothiazine-3-carboxylic acid

To a stirred solution of 4-amino-3-mercaptobenzoic acid(1.6 g, 9.31 mmol) and 2-chloro-l,3- dinitro-5-(trifluoromethyl)benzene(1.5 g,7.57 mmol) in EtOH (30 mL) was added NaOH (1.1 g,28.3 mmol) and stirring continued for 16h. The reaction mixture was diluted with water,neutralized with citric acid solution, solid was filtered, and dried by azotropic distillation using toluene to gave title compound as brown solid(1.6 g, crude). MS(ESI)355(M-H) ⁺ .

Step 3: Methyl 9-nitro-7-(trifluoromethyl)-10H-phenothiazine-3-carboxylate

To a stirred solution of 9-nitro-7-(trifluoromethyl)-10H-phenothiazine-3-carboxylic acid (0.2 g, 0.561 mmol) in MeOH (10 mL) was added conc.H ₂ SO ₄ (0.1 mL) and reaction mixture was refluxed for 2h. The reaction mixture was concentrated, residue was diluted with EtOAc, and washed with sat. NaHCC solution, dried over sodium sulphate, and concentrated to gave title compound as brown solid(0.18 g, 90%):MS(ESI)369(M-H) ⁺ .

Step 4: Methyl 9-amino-7-(trifluoromethyl)-10H-phenothiazine-3-carboxylate

To a stirred solution of methyl 9-nitro-7-(trifluoromethyl)-10H-phenothiazine-3-carboxylate (0.18 g, 0.48 mmol) were added Zn dust (0.158 g,2.43 mmol),NH ₄ Cl (0.128 g,2.43 mmol) and stirring continued for 2h. The reaction mixture was filtered and evaporated to gave title compound as yellow solid(0.165 g, quantitative):MS(ESI)339(M-H) ⁺ .

Step5: Methyl 9-((l-(2-((teri-butoxycarbonyl)amino)ethyl)piperidin-4-yl)am ino)-7- (trifluoromethyl)-10H-phenothiazine-3-carboxylate

To a stirred solution of methyl 9-amino-7-(trifluoromethyl)-10H-phenothiazine-3- carboxylate(0.25 g, 0.73 mmol)) in 1 ,2-dichloroethane (5 mL) were added tert-butyl (2-(4- oxopiperidin-l-yl)ethyl)carbamate (0.266g,1.10 mmol),AcOH(0.1 mL), sodiumborohydride(0.28 g, 7.35 mmol) and stirring continued for 12 h. The reaction mixture was washed with sodium bicarbonate solution, extracted with ethyl acetate, the combined organic phase was dried over Na ₂ S0 ₄ , concentrated, and the crude product was purified by column chromatography using 5- 10% MeOH/DCM as eluent to give title compound as yellow solid(0.12 g,29%):MS (ESI) m/z 567 (M+H) ⁺ .

Step6: Methyl 9-((l-(2-aminoethyl)piperidin-4-yl)amino)-7-(trifluoromethyl )-10H- phenothiazine-3-carboxylate

To a stirred solution of methyl 9-((l-(2-((tert-butoxycarbonyl)amino)ethyl)piperidin-4- yl)amino)-7-(trifluoromethyl)-10H-phenothiazine-3-carboxylat e(0.02 g,0.035 mmol) in dichloromethane (5mL) was added a solution of 4 N HC1 in dioxane (1 mL) at 0 °C and stirring continued at room temperature for 1 h. The reaction mixture was concentrated, dissolved in saturated sodium bicarbonate solution, extracted with dichloromethane, and concentrated. The residue was washed with DCM/«-pentane(l :10) to gave the title compound asbrown solid (0.009 g,56%y ^l a NMR (DMSO-d6, 400 MHz) δ, 1.43-1.45 (m,2H), 1.88-2.07 (m,5H), 2.30-2.32 (m,2H), 2.81 (s,3H),3.75(s,3H) 5.30 (s, 1H),6.53 (S,1H),6.59(S,1H),6.92 (d,J = 8.4Hz,lH),7.42(s,lH), 7.60(d,J= 8.4 Hz,lH),8.44 (bs,l H);MS (ESI) 467(M+H) ⁺ ; HPLC purity: 99.43%.

Figure 17 shows general reaction scheme XXIII for the synthesis of selectedl, 3, 6-trisubstituted phenothiazenes. 6-bromobenzo[d]thiazol-2-amine or 7-bromobenzo[d]thiazol-2 -amine (XXIIIa)hydrolized with KOH then Nucleophilic substitution of 2-amino 5-bromo thiophenol or 2-amino 4-bromo thiophenol /2-amino 5-bromophenols or 2-amino 4-bromo thiophenol (XXIIIb) with substituted aryl halides (XXIIIb) followed by insituSmiles rearrangement yielded substituted phenothiazene/substituted phenoxazines (XXIIIc). Compounds XXIIIc were reduced using Zn NH4C1 to yield the corresponding 1 -amino 6-bromophenothiazenes or 1 -amino 7- bromophenothiazenes /l -amino 6-bromo phenoxazines or 1 -amino 7-bromophenothiazenes (XXIIId). Reductive amination of compound XXIIId with various aldehydes or ketones yielded corresponding ^-alkylated phenothiazines ΧΧΙΠβ, which were further deprotected to give the corresponding free amines XXIIIf. and alkylated to give XXIIIg. Further protection of the XXIIIg with boc anhydride to give tri protected compounds of XXIIIh . Further Buchwald coupling of compound XXIIIh with various amines, followed by deprotection gave XXIII 1 with corresponding salts. And Further Suzuki coupling of compound of XXVIIIh with boronic acids followed by deprotection gave XXIII 3 with corresponding salts and also with XXIIIj double bond reduction with platinum oxide and followed by deprotection gave ΧΧΙΠ 2

Compound 272:N-(l-(2-aminoethyl)piperidin-4-yl)-7-(4-aminopiperidin-l -yl)-3- (trifluoromethyl)-lOH-phenothiazin-l-amine

Step 1: 2-amino-5-bromobenzenethiol:

To a stirred solution of 6-bromobenzo[d]thiazol-2-amine (100 g, 436.68 mmol, compound-1) in H ₂ 0 (1000 mL) was added potassium hydroxide (500 g, 8928.57 mmol) portion wise for a period of 30 min at 0 °C and stirred the reaction mixture at 120 °C for 12 h. The Progress of the reaction was monitored by TLC. The reaction mixture was cooled to 0 °C, neutralized with Acetic acid (pH~ 7) at 0 °C and stirred at rt. After stirring the reaction mixture for 10 min, was added toluene (2x500 mL), the combined organic layer was washed with water (500 mL), dried over anhydrous sodium sulphate and concentrated under reduced pressure below 50 °C. The crude compound was purified by adding ethanol (400 mL), stirred at reflux temperature for 20 min. The compound was cooled to room temperature, solid was filtered and dried under vacuum to afford 2-amino-5- bromobenzenethiol(50 g, yield: 56%) as yellow solid. 1H NMR (400 MHz, DMSO-d ₆ ) δ 5.7 (s, 2H), 6.85 (d, J=2.3 Hz, 1H), 7.2 (d, J = 6.3 Hz, 1H), 7.2-7.3 (d, J= 6.3 Hz 1H). LC-MS m/z (M+H): 204.0

Step 2: 7-bromo-l-nitro-3-(trifluoromethyl)-10H-phenothiazine:

To a stirred solution of 2-amino-5-bromobenzenethiol (30 g, 147.05 mmol), 2-chloro-l ,3-dinitro- 5-(trifluoromethyl)benzene (39.9 g, 147.77 mmol) in DMF (150 mL) was added sodium hydroxide (188 g, 441.15 mmol) at 0 °C and stirred the reaction mixture at room temperature for 2 h. Progress of the reaction was monitored by TLC. The reaction mixture was quenched with ice-cold water (200 mL), extracted with ethyl acetate (2x300 mL). The combined organic layer was washed with water (500 mL), dried over anhydrous sodium sulphate filtered and

concentrated under reduced pressure. The crude compound was diluted with ethanol (120 mL), stirred for 20 min solid was filtered and dried under vacuum to afford 7-bromo-l-nitro-3- (trifluoromethyl)-lOH-phenothiazine (10 g, yield: 17%) as black solid. 1H NMR (400 MHz, DMSO-i ₆ ) 7.0 (d, , J= 8.52 Hz, 1Η),7.2-7.3 (d, , J= 2.15 Hz, 2H),7.7 (s, 1H), 8.0 (s, 1H), 9.8 (s, 1H).

Step 3: 7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-amine:

To a stirred solution of 7-bromo-l-nitro-3-(trifluoromethyl)-10H-phenothiazine (11 g, 28.13 mmol) in MeOH (110 mL) water (50 mL)was added Zn powder (9.13 g, 140.46 mmol), NH ₄ C1 (7.5 g,140.44 mmol) at 0 C and stirred the reaction mixture at room temperature for 2 h. The progress of the reaction was monitored by TLC. The reaction mixture was filtered through a pad of celite; the filtrate was evaporated under reduced pressure. The residue was diluted with EtOAc (500 mL), washed with water (500 mL). The organic layer was dried over anhydrous sodium sulpahte and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (e luted with 20% EtOAc in Hexane) to afford 7-bromo-3- (trifluoromethyl)-l OH-phenothiazin-1 -amine (8 g, yield: 80%) as black solid. 1H NMR (400 MHz, DMSO-de) δ 5.4-5.6 (m, 2H), 6.5 (s, 1H), 6.7-6.8(d, J = 8.48Hz, 2H),7.2-7.3 (m, 21H), δ 7.9 - 8.0 (s, 1H). LC-MS m/z (M+H): 361.04

Step 4: tert-butyl 4-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl)amino) piperidine- 1-carboxylate:

To a stirred solution of 7-bromo-3-(trifluoromethyl)-l OH-phenothiazin-1 -amine (4 g, 11.11 mmol,) in 1 ,2-dichloroethane (100 mL) was added tert-butyl 4-oxopiperidine-l-carboxylate(4.42 g, 22.22 mmol, ) and 4A° Molecular sieves powder (10 g) at room temperature After stirring the reaction mixture for 1 h was added sodium triacetoxy boro hydride (11.77 g, 56.03 mmol). Reaction mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with sat NaHCC solution, extracted with ethyl acetate (2x70 mL). The combined organic layer was dried over anhydrous sodium sulpahte and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 20% EtOAc in Hexane) to afford tert- butyl 4-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l -yl)amino)piperidine-l-carboxylate (5 g, yield: 83% ) as green solid. 1H NMR (400 MHz, DMSO-d ₆ ) δ 1.2-1.3 (m, 2H),1.4 (s, 9H),1.9 (d, J =21.46Hz, 2H), 2.9 - 3.0 (br, 2H), 3.5 - 3.6 (m,lH), 3.8-3.9 (m, 2H), 5.2 (d, J = 7.16Hz ,1H), 6.5 (s, 1H), 6.6 (s, 1H), 6.7 (d, J= 8.42Hz, 2H), 7.2 (m, 2H), 8.0 (s,lH). LC-MS m/z (M+H): 544.0.

Step 5 : 7-bromo-N-(piperidin-4-yl)-3-(trifluoromethyl)- 1 OH-phenothiazin-1 -amine

To a stirred solution of tert-butyl 4-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l - yl)amino)piperidine-l-carboxylate (5 g, 9.19 mmol) in DCM (50 mL) was added 4M HC1 in 1,4- dioxane (10 mL) at 0 °C and stirred the reaction mixture at room temperature for 1 h. Progress of the reaction was monitored by TLC. The reaction mixture was evaporated under reduced pressure. The residue was basified with saturated NaHCC solution (PH-7 to 8), extracted with ethyl acetate (2x100 mL). The combined organic layer was dried over anhydrous sodium sulpahte and concentrated under reduced pressure to afford 7-bromo-N-(piperidin-4-yl)-3- (trifluoromethyl)-lOH-phenothiazin-l -amine (3.4 g, yield: 85%) as green solid. 1H NMR (400 MHz, DMSO-de) δ 1.2 (s, 2H), 1.3-1.4 (m, 2H), 1.9(m, 2H), 2.6-2.7(t, J=10.98Hz, 2H), 3.0 - 3.07 (m, 2H), 3.4- 3.43 (m, 1H), 5.2 (d, J=3.06Hz ,1H), 6.5(s, 1H), 6.6 (s, 1H), 6.8 (d, J = 8.43Hz, 1H), 7.2- 7.23 (m, 2H), 8.1(s,lH).LC-MS m/z (M+H): 444.0.

Step 6: tert-butyl (2-(4-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)piperidin-l-yl)ethyl)carbamate

To a stirred solution of 7-bromo-N-(piperidin-4-yl)-3-(trifluoromethyl)-10H-phenothia zin-l - amine (3.4 g, 7.67 mmol) in Acetonitrile (34 mL) was added K ₂ CO ₃ (3.17 g, 22.97 mmol) followed by tert-butyl (2-chloroethyl)carbamate (1.79 g, 9.97 mmol) at 0 °C and stirred the reaction mixture at 70 °C for 1 h. The progress of the reaction was monitored by TLC. The reaction mixture was cooled to room temperature, reaction mixture was diluted with water (20 mL), extracted with ethyl acetate (2x100 mL). The combined organic layer was dried over anhydrous sodium sulpahte and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 2-8% EtOAc in Hexane) to afford tert- butyl (2-(4-((7-bromo-3-(trifluoromethyl)-l OH-phenothiazin-1 -yl)amino)piperidin-l - yl)ethyl)carbamate (700 mg, yield: 16%) as yellow solid. 1H NMR (400 MHz, DMSO-d ₆ ) δ 1.3 (s, 1H),1.9- 2.1 (m, 2H), 2.1-2.2 (m, 2H), 3.1-3.2 (m, 2H), 3.3-3.4 (m, 5H), 5.6 (s, 1H), 6.2 (s,lH), 6.5 (s, 1H), 6.6 (d, J= 27.67Hz, 2H),7.1 (d, J = 24.63Hz, 1H),7.2( m, 2H), 8.3-8.4 (m,4H), 10.9 (s, 1H). LC-MS m/z (M+H): 587.19.

Step 7:tert-butyl 7-bromo-l-((tert-butoxycarbonyl)(l-(2-((tert-butoxycarbonyl) amino)ethyl) piperidin-4-yl) amino)-3-(trifluoromethyl)-10H-phenothiazine-10-carboxylate:

To a stirred solution of tert-butyl (2-(4-((7-bromo-3-(trifluoromethyl)-l OH-phenothiazin-1 - yl)amino)piperidin-l-yl)ethyl)carbamate (700 mg, 1.19 mmol) in Acetonitrile (10 mL) was added DMAP (364 mg, 2.98 mmol) followed byditert-butyl dicarbonate (0.78 mL,3.57 mmol) at 0 °C and stirred the reaction mixture at 80 °C for 1 h. The progress of the reaction was monitored by TLC. The reaction mixture was cooled to room temperature, reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (2x50 mL). The combined organic layer was dried over anhydrous sodium sulpahte filtered and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 2-8% EtOAc in Hexane) to afford tert-butyl 7-bromo-l-((tert-butoxycarbonyl)(l -(2-((tert-butoxycarbonyl) amino)ethyl) piperidin-4-yl) amino)-3-(trifluoromethyl)-10H-phenothiazine-10-carboxylate (700 mg, yield: 74%) as off white solid. 1H NMR (400 MHz, DMSO-d ₆ ) δ 1.3 (s, 2H),1.3 (s, 10H), 1.5(s, 19H),1.6-165(m, 2H),1.8-1.9 (m, 1H), 1.91 - 2.01(m, 1H), 2.1-2.2 (m,2H), 2.4-2.56 (m, 2H), 2.8-2.89 (m, 2H), 3.4-3.5 (m, 1H), 3.6 (t, J= 6.63Hz, 2H), 3.5-3.6 (br,lH), 6.8 (s, 1H), 6.9 (s, 1H),7.6 (d, J= 2.17Hz 1H), 7.8 -7.9 (m, 1H).LC-MS m/z (M+H): 787.2. Step 8: tert-butyl l-((tert-butoxycarbonyl)(l-(2-((tert-butoxycarbonyl)amino)et hyl)piperidin-

4-yl)amino)-7-(4-((tert-butoxycarbonyl)amino)piperidin-l- yl)-3-(trifluoromethyl)-10H- phenothiazine-10-carboxylate

To a stirred solution of tert-butyl 7-bromo-l -((tert-butoxycarbonyl)(l -(2-((tert-butoxycarbonyl) amino)ethyl) piperidin-4-yl) amino)-3-(trifluoromethyl)-10H-phenothiazine-10-carboxylate (190 mg, 0.24 mmol), tert-butyl piperidin-4-ylcarbamate (62.4 mg,0.31 mmol) in 1 4 Dioxane(4 mL) was added cessium carbonate (195 mg, 0.6 mmol) at room temperature. After degassed with argon for 10 min was added Pd ₂ (dba)3 (21.9 mg, 0.02 mmol), Xanthphos,(27.7 mg, 0.04 mmol) again degassed for 5 min and stirred the recaction mixture at 1 10 °C for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with water (10 mL), extracted with ethyl acetate (2x20 mL). The combined organic layer was dried over anhydrous sodium sulpahte and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 2-3% MeOH/DCM) to afford tert-butyl l -((tert- butoxycarbonyl)(l -(2-((tert-butoxycarbonyl)amino)ethyl)piperidin-4-yl)amino)- 7-(4-((tert- butoxycarbonyl)amino)piperidin-l -yl)-3 -(trifluoromethyl)- 1 OH-phenothiazine- 10-carboxylate (30 mg, yield: 13%) as grey solid. 1H NMR (400 MHz, OMSO-d ₆ ) δ 1.3 (s, 9H), 1.33 (s, 18H), 1.44-1.46 (m, 20H), 1.89-1.97 (m, 3H), 2.10-2.25 (m, 5H), 2.42-2.49 (m, 2H), 2.72-2.85 (m, 4H), 3.41 (brs,2H), 3.56-3.64 (m, 4H) ,6.81 -6.90 (m,2H), 6.93-6.98 (m,3H),7.51 (d, J = 8.59Hz, 1H), LC-MS m/z (M+H): 906.1

Step 9: N-(l-(2-aminoethyl)piperidin-4-yl)-7-(4-aminopiperidin-l-yl) -3-(trifluoromethyl)- 1 OH-phenothiazin-1 -amine :

To a stirred solution of tert-butyl l -((tert-butoxycarbonyl)(l -(2-((tert- butoxycarbonyl)amino)ethyl)piperidin-4-yl)amin^

yl)-3-(trifluoromethyl)-10H-phenothiazine-10-carboxylate (45 mg, 0.04 mmol)in DCM (1 mL) was added 4M HC1 in 1,4-dioxane (3 mL) at 0 °C and stirred the reaction mixture at room temperature for 2 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was washed with ether (2x2 mL) and dried under reduced pressure to afford N-(l -(2-aminoethyl) piperidin-4-yl)-7-(4-aminopiperidin-l -yl)-3- (trifluoromethyl)-l OH-phenothiazin-l -amine (20 mg, yield: 83%) as off white solid. 1H NMR (400 MHz, OMSO-d ₆ ) δ 1.93-1.99 (m, 4H), 2.1 1 (d, J =10.7Hz, 4H), 2.91 (s, 2H), 3.1 1 - 3.19 (m, 2H), 3.31 -3.39 (m, 8H), 3.61 -3.68 (m, 5H), 6.61 (d, J = 22.4Hz, 2H), 7.09 (s, 1H), 7.20- 7.43 (m, 2H), 8.32 (d, J =17.5Hz, 6H), 8.61 (s, 1H), 10.89 (s, 1H). LC-MS m/z (M+H): 507.1

Compound 359: N-(l-(2-aminoethyl) piperidin-4-yl)-7-(cyclopent-l-en-l-yl)-3-

(trifluoromethyl)-lOH-phenothiazin-l-amine hydrochloride

Step 10: tert-butyl l-((tert-butoxycarbonyl)(l-(2-((tert- butoxycarbonyl)amino)ethyl)piperidin-4-yl)amino)-7-(cyclopen t-l-en-l-yl)-3-

(trifluoromethyl)-lOH-phenothiazine-lO-carboxylate

To a stirred solution of tert-butyl 7-bromo-l -((tert-butoxycarbonyl)(l-(2-((tert- butoxycarbonyl)amino)ethyl)piperidin-4-yl)amino)-3 -(trifluoromethyl)- 1 OH-phenothiazine- 10- carboxylate (200 mg, 0.254 mmol) in 1 4 Dioxane/H ₂ 0 (8 mL) was added potassium carbonate (70mg, 0.508 mmol) at room temperature. After degassed with argon for 10 min was added cyclopent-l -en-l-ylboronic acid (58 mg, 0.508 mmol,)and was added Pd2(dppf)C12 DCM complex.(l l mg, 0.05 mmol) again degassed for 5 min and stirred the reaction mixture at 110 °C for 12 h. Progress of the reaction was monitored by TLC. The reaction mixture was filtered through a pad of celite; the filtrate was concentrated under reduced pressure. The crude compound was purified by combi-flash chromatography (eluted with 2-3% EA in Pet Ether) followed by preparative TLC to afford tert-butyl l-(l -(2-(tert-butoxycarbonylamino) ethyl) piperidin-4-ylamino)-7-cyclopentenyl-3-(trifluoromethyl)-10H -phenothiazine-10-carboxylate as sticky liquid (85 mg, yield: 62%). ¾ NMR (400 MHz, DMSO-d ₆ ) δ 1.22 (s, 9H), 1.3 (s, 18H), 1.5-1.6 (m, 3H), 1.81-1.9 (m,lH), 1.97 (t, J = 7.4Hz, 3H), 2.17-2.2 (m, 2H), 2.31 -2.39 (m ,1H), 2.43 (t, J = 6.72Hz, 2H), 2.6-2.7 (m, 2H), 2.84-2.89 (m, 2H), 3.29-3.43 (m, 1H), 3.56 (t, J = 6.5Hz, 2H), 5.42 (br, 1H), 6.34 (s, 1H), 6.84 (s, 1H), 6.96 (s, 1H), 7.46 (d, J = 8.3Hz, 1H), 7.51(s, 1H), 7.71 (d, J = 8.32 Hz, 1H). LC-MS m/z (M+H): 775

Step 11: N-(l-(2-aminoethyl) piperidin-4-yl)-7-(cyclopent-l-en-l-yl)-3-(trifluoromethyl)- 1 OH-phenothiazin-1 -amine hydrochloride

To a stirred solution of tert-butyl l-(l-(2-(tert-butoxycarbonylamino) ethyl) piperidin-4-ylamino)- 7-cyclopentenyl-3-(triflu or omethyl)-! OH-phenothiazine- 10-carboxylate (30 mg, 0.03870mmol) in CH2CI2 (1 mL) was added 1,4-dioxane HC1 (2 niL) at 0 °C and stirred the reaction mixture at rt for 3 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was triturated with n-pentane (2x2 mL) and dried n to afford N-(l -(2-aminoethyl) piperidin-4-yl)-7-(cyclopent-l -en-1 -yl)-3-(trifluoromethyl)-l 0H- phenothiazin-1 -amine hydrochloride as pale yellow solid. (14 mg, yield: 77%). ¹ H NMR (400 MHz, DMSO-de) 51.8- 1.9 (m, 3H), 2.1 (d, J = 16.1Hz, 2H), 2.4-2.5 (m, 2H), 2.5-2.6 (m, 2H), 3.1 -3.2 (m, 2H), 3.3-3.4 (m, 4H), 3.6-3.7 (m, 3H), 6.1 (s, 1H), 6.5-6.6 (m, 2H), 7.01 (s, 2H), 7.2 (d, J = 7.2Hz 1H), 8.2 (br s, 4H). LC-MS m/z (M+H): 475.2

Compound 354: N-(l-(2-aminoethyl) piperidin-4-yl)-7-cyclopentyl-3-(trifluoromethyl)-10H- phenothiazin-l-amine hydrochloride

Step 12: tert-butyl l-((tert-butoxycarbonyl)(l-(2-((tert- butoxycarbonyl)amino)ethyl)piperidin-4-yl)amino)-7-cyclopent yl-3-(trifluoromethyl)-10H- phenothiazine-10-carboxylate

To a stirred solution of tert-butyl l-((tert-butoxycarbonyl)(l -(2-((tert- butoxycarbonyl)amino)ethyl)piperidin-4-yl)amino)-7-(cyclopen t-l-en-l -yl)-3-(trifluoromethyl)- lOH-phenothiazine-10-carboxylate (50 mg, 0.06459 mmol) in Toluene (5 mL) was added platinum oxide (30 mg) at room temperature under ¾ atmosphere for 6 h. The progress of the reaction was monitored by TLC. The reaction mixture was filtered through celite bed filtrate was concentrated under reduced pressure, crude product was purified by prep TLC (30% ethyl acetate/n-Hexane) to afford tert-butyl 1 -((tert-butoxycarbonyl)(l -(2-((tert-butoxycarbonyl)amino)ethyl)piperidin-4- yl)amino)-7-cyclopentyl-3-(trifluoromethyl)-l OH-phenothiazine-10-carboxylate as colorless sticky solid (21 mg, yield: 42%). LC-MS m/z (M+H): 777.1

Step 13: N-(l-(2-aminoethyl) piperidin-4-yl)-7-cyclopentyl-3-(trifluoromethyl)-10H- pheiiothiazin-1 -amine hydrochloride

To a stirred solution of tert-butyl l -((tert-butoxycarbonyl)(l -(2-((tert- butoxycarbonyl)amino)ethyl)piperidin-4-yl)amino)-7-cyclopent yl-3-(trifluoromethyl)-l OHphenothiazine-10-carboxylate (52 mg, 0.073 mmol) in DCM (1 mL) was added 1,4-dioxane HC1 (2 mL) at 0 °C and stirred the reaction mixture at room temperature for 3 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was triturated with n-pentane (2x2 mL) and dried to afford N-(l -(2- aminoethyl) piperidin-4-yl)-7-cyclopentyl-3-(trifluoromethyl)-10H-phenot hiazin-l -amine hydrochloride as pale yellow solid (12 mg, yield: 100%). ^! H NMR (400 MHz, DMSO-d ₆ ) δ 1.4- 1.49 (m, 2H), 1.6-1.7 (m, 2H), 1.7-1.8 (m, 2H), 1.9-2.0 (m, 4H), 2.1(d, J = 13.1Hz, 2H), 2.8-2.9 (m, 1H), 3.1-3.2 (m, 2H), 3.27-3.33 (m, 4H), 3.6 (d, J= 11.8Hz, 3H), 5.6 (br s, 1H), 6.6 (s, 1H), 6.7 (s, 1H), 6.8 (s, 1H), 6.9(d, J = 8.4Hz, 2H), 8.2(br s, 4H), 10.94 (br s, 1H). LC-MS m/z (M+H): 477.2

Some examples of compounds synthesised by the method of Scheme XXIII are provided in Table

XXIII

Table XXIII

Figure 18 shows general reaction scheme XXIV for the synthesis of selected 1 , 3, 6-trisubstituted phenothiazenes. Boc protection of 2-aminoethyl)ethane-l ,2-diamine (XXIVa) with alkylation with bromo ethanol (XXIVb) followed by mestylation yielded diboc protected 2-(bis(2- aminoethyl)amino)ethyl methanes ulfonate to yield XXIVd. Compound XXIVd were n-alkylated with XXIIIf yielded corresponding ^-alkylated phenothiazines XXIVe, Further protection of the XXIVewith boc anhydride to give tri protected compounds of XXIVf . Further Buchwald coupling of compound XXIVg with various amine, followed by deprotection gave XXIVwith corresponding salts.

Compound 335: Nl-(2-aminoethyl)-Nl-(2-(4-((7-(pyrrolidin-l-yl)-3-(trifluor omethyl)-10H- phenothiazin-l-yl)amino)piperidin-l-yl)ethyl)ethane-l,2-diam ine hydrochloride

Step 1: di-tert-butyl (azanediylbis(ethane-2,l-diyl))dicarbamate

To a stirred solution of imidazole (5 g, 73.52 mmol) in DCM (5 mL) was added di tert-butyl dicarbonate (15.25 g, 69.95 mmol) at room temperature and stirred for 2 h. After completion of the reaction, diluted with 100 mL of DCM and washed with water (50 mL) and organic layer was dried over sodium sulfate, filtered and concentrated to give crude residue to this was added Nl-(2-aminoethyl)ethane-l,2-diamine (3.5 mL, 33.98 mmol) and stirred for 1 h at room temperature. After completion of the reaction, diluted with water (10 mL) extracted with DCM (2 X 20 mL). Combined organic layers were dried over sodium sulfate, filtered and evoparted to give crude residue, which was purified by gradient chromatography (Product eluted with 5% MeOH/DCM) to afford di-tert-butyl (azanediylbis(ethane-2,l-diyl))di carbamate as a colorless liquid (4.3 g, 40%). 1H NMR (400 MHz, CDC1 ₃ ) δ 1.44 (s, 18H), 2.73 (t, J =5.79 Hz, 4H), 4.9 (s, 1H). Step 2: di-tert-butyl (((2-hydroxyethyl)azanediyl)bis(ethane-2,l-diyl))dicarbamate

To a solution of di-tert-butyl (azanediylbis(ethane-2, 1 -diyl))dicarbamate (3.7 g, 12.19 mmol) in acetonitrile (40 mL) was added sodium carbonate (12.92 g, 122.88 mmol) at room temperature and stirred for 10 min., then 2-bromo ethanol (3.81 g) was added to the reaction mixture and stirred at 70 °C for 16 h. After completion of the reaction, diluted with water (10 mL) extracted with ethyl acetate (2 X 50 mL). The combined organic layers were dried over sodium sulfate , filtered and concentrated to give crude residue, which was purified by gradient chromatography (product eluted with 5 % of Methanol/DCM) to give di-tert-butyl (((2- hydroxyethyl)azanediyl)bis(ethane-2,l -diyl))dicarbamate as colorless liquid (3.8 g, 90%). 1H NMR (400 MHz, OMSO-d ₆ ) δ 1.28 (s, 18H), 2.41 -2.49 (m, 6H), 2.90-2.95 (m, 4H), 3.34-3.38 (m, 2H)

Step 3: 2-(bis(2-((tert-butoxycarbonyl)amino)ethyl)amino)ethyl methanesulfonate

To a stirred solution of di-tert-butyl (((2-hydroxyethyl) azanediyl) bis (ethane-2, 1 -diyl)) dicarbamate (0.75 g, 2.158 mmol) in DCM (15 mL) was added triethyl amine (0.45 g, 4.447 mmol) and mesyl chloride (0.3 g, 2.589 mmol) at 0 °C. Reaction mixture was stirred at room temperature for lh. After completion of the reaction, diluted with ice water (15 mL) and extracted with DCM (2 X 20 mL). Combined organic layers was washed with saturated sodium bi carbonate (10 mL) and followed by water (10 mL). Organic layer was dried over sodium sulfate, filtered and evaporated to give 2-(bis (2-((tert-butoxycarbonyl) amino) ethyl) amino) ethyl methanesulfonate as brown liquid (crude, 0.8 g), which was used for the next without further purification.

Step 4: di-tert-butyl (((2-(4-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)piperidin-l-yl)ethyl)azanediyl)bis(ethane-2,l-diyl) )dicarbamate

To a stirred solution of 7-bromo-N-(piperidin-4-yl)-3-(trifluoromethyl)-10H-phenothia zin-l- amine (0.2 g, 0.45 mmol) in Acetonitrile (12 mL) was added potassium carbonate (0.075 g, 0.542 mmol) stirred for lOmin., room temperature. To this 2-(bis (2-((tert-butoxycarbonyl) amino) ethyl) amino) ethyl methanesulfonate (0.235 g, 0.552 mmol) was added and stirred at room temperature for 16 h. After completion of the reaction, diluted with ethyl acetate (25 mL) and washed with water (10 mL). Organic layer was dried over sodium sulfate, filtered and evoparted to give crude residue, which was purified by gradient chromatography (product eluted with 4% methanol/DCM) to give di-tert-butyl (((2-(4-((7-bromo-3-(trifluoromethyl)-l OHphenothiazin-1 -yl)amino)piperidin-l -yl)ethyl)azanediyl)bis(ethane-2,l -diyl))dicarbamate as blue solid (65 mg, 18.6 %). 1H NMR (400 MHz, OMSO-d ₆ ): δ 1.4 (s, 23H), 1.4-1.5 (m, 2H), 1.9 (m, 2H), 2.1 (m, 2H), 2.3 (m, 1H), 2.8-2.9 (m, 2H), 2.9-3.0 (m, 5H), 5.2 (br, 1H), 6.6(m, 4H),6.8 (db, J = 8.29 Hz, 1H ), 7.2 (m, 2H), 8.1 (s, 1H)

Step 5: tert-butyl l-((l-(2-(bis(2-((tert-butoxycarbonyl)amino)ethyl)amino)ethy l)piperidin- 4-yl)amino)-7-bromo-3-(trifluoromethyl)-10H-phenothiazine-10 -carboxylate

To stirred solution of di-tert-butyl (((2-(4-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)piperidin-l-yl)ethyl)azanediyl)bis(ethane-2,l -diyl))dicarbamate (0.3 g, 0.387 mmol) in Acetonitrile (15 mL) was added 4-dimethyl amine pyridine (0.165 g, 1.356 mmol) and di tert- butyl dicarbamate (0.422 g, 1.938 mmol). Reaction mixture was stirred at room temperature for 3 h. After completion of the reaction, diluted with ethyl acetate (25 mL) and washed with water (10 mL). Organic layer was dried over sodium sulfate, filtered and evaporated to give crude product, which was purified a by gradient chromatography (product eluted with 3% methanol/DCM) to give tert-butyl 1 -((1 -(2-(bis(2-((tert- butoxycarbonyl)amino)ethyl)amino)ethyl)piperidin-4-yl)amino) -7-bromo-3-(trifluoromethyl)- lOH-phenothiazine-10-carboxylate as brown solid (0.104 g, 30 %). ^X H NMR (400 MHz, DMSO- d ₆ ): δ 1.3 (s, 29H), 1.4 (d, J = 3.29 Hz, 3H ),1.5- 1 -57 (m, 1H), 1.7-1.9 (m, 2H), 2.1 - 2.2 (m, 2H), 2.3- 2.39 (m, 2H), 2.4-2.6 (m, 6H), 2.8- 2.89 (m, 1H), 2.9-3.01 (m, 1H), 3.01 - 3.2 (m, 4H ), 3.4 - 3.45 (m, 1H), 6.6 - 6.79 (m, 1H), 6.8(s, 1H ), 6.9 (s, 1H), 7.5-7.6 (dd, J = 1.8 Hz, 1H ), 7.8 (m,2H)

Step 6: tert-butyl l-((l-(2-(bis(2-((tert-butoxycarbonyl)amino)ethyl)amino)ethy l)piperidin- 4-yl)amino)-7-(pyrrolidin-l-yl)-3-(trifluoromethyl)-10H-phen othiazine-10-carboxylate

To a stirred solution of tert-butyl 1 -((1 -(2-(bis(2-((tert- butoxycarbonyl)amino)ethyl)amino)ethyl)piperidin-4-yl)amino) -7-bromo-3-(trifluoromethyl)- lOH-phenothiazine-10-carboxylate (0.080 g, 0.0916 mmol), Pyrrolidine (13 mg,0.183 mmol) in 1, 4 Dioxane (3 mL) was added sodium hydroxide (9.1 mg, 0.229 mmol) in water (0.5 mL) at room temperature. After degassed with argon for 10 min was added Pd ₂ (dba)3 (8.39 mg, 0.009 mmol), tert butyl Xphos,(5.82 mg, 0.0137 mmol) again degassed for 5 min and stirred the reaction mixture at 110 °C for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with water (10 mL), extracted with ethyl acetate (2x20 mL). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 6% MeOH/DCM) to afford tert-butyl 1 -((1 -(2-(bis(2-((tert- butoxycarbonyl)amino)ethyl)amino)ethyl)piperidin-4-yl)amino) -7-(pyrrolidin-l-yl)-3- (trifluoromethyl)-10H-phenothiazine-10-carboxylate(35 mg, yield: 43.75%) as brown solid. 1H 5 NMR (400 MHz, OMSO-d ₆ ): δ 1.3 (s, 28H), 1.5 -1.59 (m, 1H ), 1.6- 1.7 (m, 1H), 1.8-2.0 (m, 6H), 2.1 -2.2 (m, 2H), 2.3 -2.39 (m, 1H), 2.4-2.6 (m, 5H), 2.7-2.9 (m, 2H), 2.9-3.0 (m, 4H), 3.2 - 3.3 (m, 4H ), 3.4 -3.45 (m, 1H), 5.2 (br, 1H), 6.5 - 6.61 (m, 2H ), 6.62- 6.65 (m, 2H), 6.8 (s, 1H ),6.9 (s, 1H), 7.4(d, J = 0.0216 Hz, 1H )

Step 7: Nl-(2-aminoethyl)-Nl-(2-(4-((7-(pyrrolidin-l-yl)-3-(trifluor omethyl)-10H- 10 phenothiazin-l-yl)amino)piperidin-l-yl)ethyl)ethane-l,2-diam ine hydrochloride

To a stirred solution of tert-butyl l-((l-(2-(bis(2-((tert- butoxycarbonyl)amino)ethyl)amino)ethyl)piperidin-4-yl)amino) -7-(pyrrolidin-l-yl)-3- (trifluoromethyl)-lOH-phenothiazine-lO-carboxylate (35 mg, 0.04 mmol)in DCM (0.3 rriL) was

15 added 4M HC1 in 1 ,4-dioxane (1 mL) at 0 °C and stirred the reaction mixture at room temperature for 1 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was washed with n-Pentane (5 mL) and dried under reduced pressure to afford Nl-(2-aminoethyl)-Nl-(2-(4-((7-(pyrrolidin-l-yl)-3-(trifluor omethyl)- 10H-phenothiazin-l-yl)amino)piperidin-l -yl)ethyl)ethane-l,2-diamine hydrochloride(25 mg, yield:

20 100 %) as grey solid. 1H NMR (400 MHz, OMSO-d ₆ ): δ 1.8-2.2 (m, 6H), 2.7- 2.79 (m, 3H ), 2.8 - 2.9 (m, 2H), 3.0 - 3.09 (m, 4H), 3.1- 3.2 (m, 1H), 3.3-3.5 (m, 4H), 3.6-3.7 (m, 2H), 6.5-7.4 (m, 2H), 8.2 (br, 5H),10.3 (br, 1H )

Some examples of compounds synthesised by the method of Scheme XXIV are provided in Table XXIV

25 Table XXIV Cmpd # R ^b Cmpd #

Figure 19 shows general reaction scheme XXV for the synthesis of selectedl, 3, 6-trisubstituted phenothiazenes. Buchwald coupling of nitro bromo phenothiazines (XXVa) yielded substituted phenothiazene/substituted phenoxazines (XXVb). Compounds XXVb were redusing Zn/NH4C1 to yield the corresponding 1 -amino 6-substiutted phenothiazenes/1 -amino 6-substituted phenoxazines (XXVc). Compounds XXVc were reacted with acid chlorides or acids to form corresponding amides XXVd, further deprotection gave XXVe with corresponding salts and XXVd reduction with boran DMS to give XXVf, further deprotection gave XXV with corresponding salts.

Compound 300: N-((3-aminocyclohexyl)methyl)-7-(pyrrolidin-l-yl)-3-(trifluo romethyl)-10H- phenothiazin-l-amine:

Step-1: Synthesis of tert-butyl l-nitro-7-(pyrrolidin-l-yl)-3-(trifluoromethyl)-10H- phenothiazine- 10-carboxylate:

To a stirred solution of tert-butyl 7-bromo-l-nitro-3-(trifluoromethyl)-10H-phenothiazine-10- carboxylate (lg, 2.55mmol) and pyrrolidine (907 mg, 12.78 mmol) in 1,4-Dioxan (20 mL) was added cessium carbonate (2.49 g, 7.67 mmol) at room temperature , then degassed with argon for 15min. Then added xantphos (295mg, 0.511 mmol) and Pd ₂ (dba) 3 (234mg, 0.255 mmol) degased for 5min. The reaction mixture was stirred at 110°C for 12h. The progress of the reaction was monitored by TLC. Reaction mixture was cooled to room temperature, filtered through celite bed washed with ethyl acetate (lOOmL). Filtrate was concentrated under reduced pressure. The crude compound was purified by column chromatography (silica gel 60-120 mesh, eluted with 2-3% EtOAc in pet ether) to afford tert-butyl l-nitro-7-(pyrrolidin-l-yl)-3- (trifluoromethyl)-lOH-phenothiazine-lO-carboxylate (850 mg, yield: 87%) as brown solid.

1H NMR (400 MHz, DMSO-d6) δ 1.32 (s, 9H), 1.94-1.96 (m, 4H), 3.22-3.30 (m, 4H), 6.59-6.61 (m, 2H), 7.33-7.37 (m, 1H), 7.45-7.46 (m, 1H), 7.70 -7.80 (m, 1H), 8.27 (brs, 1H), 8.32 (s,lH). LC-MS m/z (M+H): 482.1

Step-2: Synthesis of tert-butyl l-amino-7-(pyrrolidin-l-yl)-3-(trifluoromethyl)-10H- phenothiazine- 10-carboxylate:

To a stirred solution of tert-butyl l -nitro-7-(pyrrolidin-l-yl)-3-(trifluoromethyl)-10H- phenothiazine-10-carboxylate ( 850 mg, 1.76 mmol, step-1) in 1 ,4-Dioxan/H20 (lOmL, 7:3) was added Zn (918mg, 14.13 mmol) followed by ammonium chloride (791 mg, 14.13 mmol) at 0°C. Then reaction mixture was stirred at rt for 15 h.The progress of the reaction was monitored by TLC. The reaction mixture was filtered through a pad of celite, washed with ethyl acetate (100 mL). Filtrate was washed with brine solution (50 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure to afford tert-butyl 1 -amino-7- (pyrrolidin-l-yl)-3-(triflu or omethyl)-10H-phenothiazine-l 0-carboxylate (700 mg, yield: 88%) as brown solid. 1H NMR (400 MHz, DMSO-d6) δ 1.34 (s, 9H), 1.94-1.97 (m, 4H), 3.20-3.30 (m, 4H), 5.73 (s, 2H), 6.47-6.57 (m, 2H), 6.88 (s, 1H), 6.93 (s, 1H), 7.37 (d, J = 8.6Hz, 1H) LC-MS (m/z) (M+l) 452.1

Step-3: Synthesis of tert-butyl l-(3-(tert-butoxycarbonylamino)cyclohexanecarboxamido)-7- (pyrrolidin-1 -yl)-3-(trifluoromethyl)- 1 OH-phenothiazine- 10-carboxylate:

To a stirred solution of 3 -(tert-butoxycarbonylamino)cyclohexanecarboxylic acid (65 mg, 0.266 mmol) in pyridine (2.5 mL) was cooled to 0°C, then added tert-butyl l -amino-7-(pyrrolidin-l- yl)-3-(trifluoromethyl)-10H-phenothiazine-10-carboxylate (100 mg, 0.221 mmol) in pyridine (2.5 mL) was stirred at 0-10°C for lh. The progress of the reaction was monitored by TLC. The reaction mixture was poured into ice-cold water (50 mL) slowly drop wise very carefully then extracted with ethyl acetate (2x40 mL). The combined organic layer was washed with saturated NaHCC solution (100 mL) dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by column chromatography (silica gel 60- 120 mesh, eluted with 2-3% EtOAc in DCM) to afford tert-butyl l -(3-(tert- butoxycarbonylamino)cyclohexanecarboxamido)-7-(pyrrolidin-l- yl)-3-(trifluoromethyl)-10H- phenothiazine-10-carboxylate (30 mg, yield: 16%) as grey color solid. IH NMR (400 MHz, DMSO-d6) δ 1.30-1.32 (m, 9H), 1.35-1.39 (m, 13H), 1.41 -1.49 (m, 10H), 1.50-1.59 (m, 2H), 1.94-1.97(m, 7H), 3.17-3.19(s, 4H), 4.19-4.20 (m, IH), 6.10-6.18 (m, 2H), 6.54 (d, J =7.43Hz, lH),6.80-6.89 (m,lH),6.98-6.99 (m,lH),7.34-739(m,lH), 7.51-7.53 (m,lH) 7.61 - 7.17 (m,lH), 8.03 - 8.20 (m, IH). LC-MS (m/z) (M+l):677.2

Step-4: Synthesis of 3-amino-N-(7-(pyrrolidin-l-yl)-3-(trifluoromethyl)-10H-pheno thiazin-l- yl)cyclohexanecarboxamide:

To a stirred solution of tert-butyl l-(3-(tert-butoxycarbonylamino)cyclohexanecarboxamido)-7- (pyrrolidin-l-yl)-3-(trifluoromethyl)-10H-phenothiazine-10-c arboxylate (12 mg, 0.017mmol, ) in DCM (0.5 mL) was added 4M HC1 in 1,4-Dioxan (lmL) at 0°C, then the reaction mixture was stirred at rt for lh. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure obtained solid was washed with diethyl ether (2x3 mL) and dried under reduced pressure to afford 3-amino-N-(7-(pyrrolidin-l-yl)-3-(trifluoromethyl)-10H- phenothiazin-l-yl)cyclohexanecarboxamide (5 mg, yield: 45%) as grey solid. 1H NMR (400 MHz, DMSO-i/ ₆ ) δ 1.12-1.37 (m, 2H), 1.41 -1.50 (m, 1H), 1.80-1.92 (m, 8H), 2.08 (d, J = 12.1Hz, 1H), 3.03-3.20 (m, 6H), 6.20-6.27 (m, 1H), 6.29-6.32 (m, 1H), 6.74 (s, 1H), 7.08 (s, 1H), 7.27 (s, 1H), 7.93 (s, 5H), 9.67 (brs, 1H) LC-MS (m/z) (M+l) 477.2

Step-5: Synthesis of tert-butyl l-((3-(tert-butoxycarbonylamino)cyclohexyl)methylamino)-7- (pyrrolidin-l-yl)-3-(trifluoromethyl)-10H-phenothiazine-10-c arboxylate:

To a stirred solution of tert-butyl l-(3-(tert-butoxycarbonylamino)cyclohexanecarboxamido)-7- (pyrrolidin-l-yl)-3-(trifluoromethyl)-10H-phenothiazine-10-c arboxylate (130 mg, 0.19 mmol, step-3,) in THF (2 mL) was cooled to 0°C, then added B¾.DMS (2.5 mL ), reaction mixture was stirred at 60°C for lh. The progress of the reaction was monitored by TLC. The reaction mixture was cooled to rt, quenched with IN HCl (10 mL) slowly drop wise then extracted with ethyl acetate (2x30 mL) and washed with water (1x50 mL). The organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by prepative TLC (eluted with 20% EtoAc in petether) to afford tert-butyl l-((3-(tert- butoxycarbonylamino)cyclohexyl)methylamino)-7-(pyrrolidin-l -yl)-3-(trifluoromethyl)-10H- phenothiazine-10-carboxylate (20 mg, yield: 15.7%) as yellow solid.1H NMR (400 MHz, DMSO-d6) δ 1.22-1.24 (m, 8H), 1.27 (s, 9H), 1.32 (s, 9H), 1.67-1.76 (m, 4H), 1.94-2.1 (m,6H), 2.99-3.1 (m, 2H), 3.20-3.29 (m, 5H), 5.81 (d, J = 6.92Hz, 1H), 6.12-6.18 (m, 1H), 6.41-6.45 (m, 2H), 6.90 (s, 1H), 7.44 (d, J= 8.81Hz, 1H). LC-MS (m/z) (M+l) 663.2

Step-6: Synthesis of N-((3-aminocyclohexyl)methyl)-7-(pyrrolidin-l-yl)-3-(trifluo romethyl)- lOH-phenothiazin-l-amine:

To a stirred solution of tert-butyl l-((3-(tert-butoxycarbonylamino)cyclohexyl)methylamino)-7- (pyrrolidin-l-yl)-3-(trifluoromethyl)-10H-phenothiazine-10-c arboxylate (20 mg, 0.030 mmol, step- 5) in DCM (1 mL) was added 1,4-Dioxan.HCl (lmL, 4M) at 0°C, then the reaction mixture was stirred at rt for lh. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure obtained solid was washed with diethyl ether (2x3 mL) and dried under reduced pressure to afford N-((3-aminocyclohexyl)methyl)-7-(pyrrolidin-l-yl)-3- (trifluoromethyl)-lOH-phenothiazin-l -amine hydrochloride (9 mg, yield: 56%) as yellow solid. 1H NMR(400 MHz, DMSO-d6) δ 0.82 - 0.90 (m, 1H), 0.95 -1.05 (m, 1H), 1.10 -1.35 (m, 3H), 1.68- 1.88 (m, 4H), 1.89 - 2.0 (m, 3H), 2.05 - 2.15 (m, 2H), 2.95 - 3.05 (m, 2H), 3.10 - 3.25 (m, 2H), 3.85 - 4.10 (m, 3H), 6.02 - 6.90 (m, 2H), 7.93 (brs, 3H). LC-MS (M+l) m/z: 463.1

Some examples of compounds synthesised by the method of Scheme XXV are provided in Table XXV

Table XXV

Figure 20 shows general reaction scheme XXVI for the synthesis of selectedl, 3, 6-trisubstituted phenothiazenes. XXVIa reacted with acid chloride yielded corresponding amide of 1 ,3 6 tri substituted phenothiazene/substituted phenoxazines (XXVIb). Compounds XXVIb futher udergoes alkylation in microwave gave XXVIc. Compounds XXVIc deprotection of Boc to give corresponding salt XXVTd by using HC1 in dioxane and XXVIc reduce the amide bond and followed by boc deprotections gave XXVIe with corresponding salts

Compound 298: Nl-(2-aminoethyl)-N3-(7-bromo-3-(trifluoromethyl)-10H-phenot hiazin-l- yl)propane-l,3-diamine

Step 1: N-(7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl)-3-chlo ropropanamide

To a stirred solution of 7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l -amine (2 g, 5.115 mmol) in DCM (20 mL) was added pyridine (5.2 mL) and 3-chloropropanoyl chloride (0.8 g, 6.138 mmol) at 0 °C. Reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, solvent was removed under vacuo to give crude residue, which was poured on to ice and stirred for 0.5 h, solid was collected from filration and washed with water (10 mL) dried to give N-(7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl)-3-chlo ropropanamide as light green solid (1.1 g, 47%)1H NMR (400 MHz, DMSO-d6) δ 2.89 (t, J = 6.64Hz, 3H), 3.38 (t, J = 6.72Hz, 2H), 6.83 (d, J = 8.24Hz,lH), 7.19-7.25 (m, 3H), 7.31(brs, 1H), 8.01 (s, 1H), 9.58 (s, 1H).LC-MS m/z (M+H): 452.11

Step 2: tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl)a mino)-3- oxopropyl)amino)ethyl)carbamate

To a stirred solution of N-(7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl)-3- chloropropanamide (0.5 g, 1.108 mmol) in 1 ,4-Dioxan (5 mL) was added tert-butyl (2- aminoethyl)carbamate (0.212 g, 1.33 mmol). Reaction mixture was stirred at 120 °C in Microwave irradiation for 0.5 h. After completion of the reaction, solvent was removed under vacuo to give crude residue, which was poured on to DCM (5 mL), solid was filtered and dried to give tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl)a mino)-3- oxopropyl)amino)ethyl)carbamateas green solid (0.4 g, 63%). 1H NMR (400 MHz, DMSO-d6) δ 1.22 (m, 2H), 1.32 (s 9H), 2.55-2.64 (m, 2H), 2.82-2.88 (m, 2H), 2.97-3.05(m, 2H), 6.72 (brs, 1H), 6.78(d, J = 8.19Hz, 1H), 7.20 (d, J = 8.1Hz,3H),7.28 (s, 1H), 8.13 (brs, 1H).LC-MS m/z (M+H): 577

Step 3: 3-((2-aminoethyl)amino)-N-(7-bromo-3-(trifluoromethyl)-10H-p henothiazin-l- yl)propanamide

To a stirred solution of tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)-3-oxopropyl)amino)ethyl)carbamate (100 mg, 0.21 mmol) in DCM (2 mL) was added 4M HC1 in 1,4-Dioxan (0.5 mL) at 0 °C, then the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, reaction mixture was concentrated under reduced pressure to give solid which was washed with diethyl ether (2x3 mL) and n-Pentane (2 mL) , dried under reduced pressure to afford 3-((2-aminoethyl)amino)-N-(7-bromo-3- (trifluoromethyl)-10H-phenothiazin-l -yl)propanamide hydrochloride ( 50 mg, yield: 43 %) as brown solid. 1H NMR (400 MHz, DMSO-i/ ₆ ) δ 2.96-2.99 (m, 2H), 3.12-3.30 (m, 7H), 7.06 (d, J = 8.19Hz, 1H), 7.15-7.23 (m, 3H), 7.48 (s, 1H), 8.22 (brs, 3H), 8.67 (s, 1H), 9.35 (brs, 2H), 10.07 (s, 1H) LC-MS (M+l):475.12

Step 4: tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)propyl)amino)ethyl)carbamate (BI-001-0027-150)

To a stirred solution of tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl) amino)-3-oxopropyl) amino) ethyl) carbamate (400 mg, 0.695 mmol) in THF (5 mL) was added 2M borane.DMS in THF solution at 0°C. Reaction mixture was stirred at 50 °C for 2 h. After completion of the reaction quenched with ice and extracted with ethyl acetate (2 X 25 mL). The combined organic layers were dried over sodium sulfate, filtered and evaporated to give crude residue, which was purified by gradient column chromatography (product eluted with 3% methanol in DCM) to afford tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)propyl)amino)ethyl)carbamate as Brown Solid (130 mg, 33%). 1H NMR (400 MHz, DMSO-i/ ₆ ) δ 1.22 (s, 4H), 1.37 (s, 9H),1.62-1.85 (m, 2H), 2.62-2.70 (m, 2H), 2.71-2.78 (m, 2H), 3.02-3.10 (m, 2H), 3.11-3.19 (m, 2H), 6.53 (m, 2H), 6.78 - 6.82 (m, 2H), 7.19-7.24 (m, 32H), 8.12 (s, 1H). LC-MS m/z (M+H): 563

Step 5: Nl-(2-aminoethyl)-N3-(7-bromo-3-(trifluoromethyl)-10H-phenot hiazin-l- yl)propane-l,3-diamine (BI-001-0027-167)

To a stirred solution of tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l- yl)amino)propyl)amino)ethyl)carbamate (50 mg, 0.089 mmol) in DCM (2 mL) was added 4M HC1 in 1,4-Dioxan (1 mL) at 0°C, then the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, reaction mixture was concentrated under reduced pressure to give solid which was washed with diethyl ether (3 mL) and n-Pentane (3 mL) , dried under reduced pressure to afford Nl -(2-aminoethyl)-N3-(7-bromo-3-(trifluoromethyl)-10H- phenothiazin-l -yl)propane- 1 ,3 -diamine hydrochloride ( 30 mg, yield: 68 %) as yellow solid. 1H NMR (400 MHz, OMSO-d ₆ ) δ 1.97-2.04 (m, 2H), 3.10-3.27 (m, 8H), 3.1 1 -3.24 (m, 8H), 6.52- 6.58 (m, 2H), 7.10-7.20 (m, 2H), 8.25 (brs, 3H), 8.67 (s, 1H) 9.22 (brs, 2H) LC-MS (M+l) 461.2

Some examples of compounds synthesised by the method of Scheme XXVI are provided in Table XXVI

Table XXVI

Figure 21 shows general reaction scheme XXVII for the synthesis of selectedl , 3, 6-trisubstituted phenothiazenes. XXVc reacted with acid chloride yielded corresponding amide of 1 ,3 6 tri substituted phenothiazene/substituted phenoxazines (XXVIIa). Compounds XXVIIafuther udergoes alkylation in microwave gave XXVIIb. Compounds XXVIIb reduce the amide bond and followed by boc deprotections gave XXVII with corresponding salts

Compound 315 :-3-((2-aminoethyl)amino)-N-(7-(pyrrolidin- l-yl)-3-(trifluoromethyl)-l 0H- phenothiazin-l-yl)propanamide hydrochloride

Step l:tert-butyl l-(3-chloropropanamido)-7-(pyrrolidin-l-yl)-3-(trifluorometh yl)-10H- phenothiazine-10-carboxylate

To a stirred solution of tert-butyl l-amino-7-(pyrrolidin-l -yl)-3-(trifluoromethyl)-10H- phenothiazine-10-carboxylate (0. 65 g, 1.441 mmol) in DCM (20 mL) was added pyridine (2 mL) and 3-chloropropanoyl chloride (0.218 g, 1.729 mmol) at 0 °C. Reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, solvent was removed under vacuo to give crude residue, diluted with ethyl acetate (50 mL) washed with water (50 mL). Organic layer was dried over sodium sulfate, filtered and evaporated to give crude product, which was purified by gradient column chromatography (20 -30 % ethyl acetate/Hexane) to give tert-butyl l-(3-chloropropanamido)-7-(pyrrolidin-l -yl)-3-(trifluoromethyl)-10H-phenothiazine- 10-carboxylate as off white solid (0.7 g, 89%)1H NMR (400 MHz, DMSO-d6) δ 1.29 (s, 9H), 1.90 -1.95 (m, 4H), 2.90 - 3.10 (m, 2H), 3.18 - 3.22 (m, 4H), 3.90 (t, J = 6.72Hz, 2H), 6.52-6.59 (m, 2H), 7.52 (d, J = 8.24Hz, 1H), 7.62(s, 1H), 8.14 (brs, 1H), 9.77 (brs, 1H).LC-MS m/z (M+H): 542.11

Step 2: tert-butyl l-(3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)propanamido )-7- (pyrrolidin-l-yl)-3-(trifluoromethyl)-10H-phenothiazine-10-c arboxylate

To a stirred solution of tert-butyl l -(3-chloropropanamido)-7-(pyrrolidin-l-yl)-3- (trifluoromethyl)-lOH-phenothiazine-lO-carboxylate (0.7 g, 1.29 mmol) in 1,4-Dioxan (3 mL) was added tert-butyl (2-aminoethyl)carbamate (0.250 g, 1.549 mmol). Reaction mixture was stirred at 120 °C in Microwave irradiation for 0.5 h. After completion of the reaction, solvent was removed under vacuo to give crude residue, which was diluted with ethyl acetate ( 20 mL) and washed with water (20 rriL). Organic layer was dried over sodium sulfate, filtered and evaporated to give crude product. Crude product was purified by gradient column chromatography (3 -4 % methanol/DCM) to givetert-butyl l-(3-((2-((tert-butoxycarbonyl) amino) ethyl) amino) propanamido)-7-(pyrrolidin-l -yl)-3-(trifluoromethyl)-10H-phenothiazine- 10-carboxylateas off white solid (0.62 g, 72%). 1H NMR (400 MHz, DMSO-d6) δ 1.30 (s, 9H), 1.37 (s,9H) 1.95 (m, 4H), 2.50-2.55 (m, 2H), 2.65-2.70 (m, 2H), 2.81 -2.89 (m, 2H), 3.17- 3.25(m, 4H), 6.51-6.59 (m, 2H), 6.80 (brs , 1H), 7.45 (d, J = 8.1Hz,lH), 7.58 (s, 1H), 8.45 (brs, 1H). LC-MS m/z (M+H): 666.30

Step 3: 3-((2-aminoethyl)amino)-N-(7-(pyrrolidin-l-yl)-3-(trifluorom ethyl)-10H- phenothiazin-l-yl)propanamide hydrochloride

To a stirred solution of tert-butyl l-(3-((2-((tert-butoxycarbonyl) amino) ethyl) amino) propanamido)-7-(pyrrolidin-l-yl)-3-(trifluoromethyl)-10H-phe nothiazine-10-carboxylate (100 mg, 0.15 mmol) in DCM (10 mL) was added 4M HC1 in 1,4-Dioxan (2 mL) at 0 °C, then the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, reaction mixture was concentrated under reduced pressure to give solid which was washed with diethyl ether (3 mL) and n-Pentane (3 mL), dried under reduced pressure to afford 3-((2- aminoethyl)amino)-N-(7-(pyrrolidin-l -yl)-3-(trifluoromethyl)-l OH-phenothiazin-1 - yl)propanamide hydrochloride (60 mg, 75 %) as grey solid. ^X H NMR (400 MHz, DMSO-i/ ₆ ) δ 1.92 (brs, 4H), 3.02 (brs, 3H), 3.15-3.35 (m, 9H), 6.20-6.47 (m, 2H), 6.80-7.01 (m, 1H), 7.02- 7.20 (m, 1H), 7.21-7.42 (m, 1H), 8.25 (s, 4H), 9.45 (s, 2H), 10.01 (brs, 1H) LC-MS (M+l):466.38

Some examples of compounds synthesised by the method of Scheme XXVII are provided Table XXVII Cmpd # R Cmpd # R R ^b

Figure 22 shows general synthetic scheme XXVIII for the synthesis of selectedl, 3, substituted phenothiazenes. Nucleophilic substitution of 2-amino thiophenol (XXVIIIa) with aryl halides (XXVIIIb)followed by insituSmiles rearrangement yielded 1,3-disubstituted phenothiazenes (XXVIIIc), which are reduce by Zn/NH4C1 to yield the corresponding 1 -amino subtituted phenothiazenes (XXVIIId). Reductive amination of compound XXVIIId with Keto yielded corresponding ^-alkylated phenothiazines XXVIIIg, Compounds XXVIIIg protected with boc yieldedtri boc of XXVIIIh was hydrolysed to gave acid (XXVIIIi) , XXXVIIIi were reacted with acid chlorides or acids to form corresponding amides XXVIIIj, which further deprotected to yield corresponding title compounds XX VEIL

Compound 288: (l-(l-(2-aminoethyl)piperidin-4-ylamino)-10H-phenothiazin-3- yl)(pyrrolidin-l-yl)methanone:

Ste -1: Synthesis of methyl 4-chloro-3,5-dinitrobenzoate:

To a stirred solution of 4-chloro-3,5-dinitrobenzoic acid (lOg, 40.5 mmol) in MeoH (100 mL) was added concentrated sulphuric acid( 5 mL) at 0°C, then the reaction mixture was stirred at 80°C for 8 h. The progress of the reaction was monitored by TLC. Reaction mixture was concentrated under reduced pressure remove the solvent, obtained residue was diluted with ice- water (200 mL). Then extracted with ethyl acetate (2x100 mL), the combined organic layer was washed with saturated. NaHCC solution (2x100 mL).The organic layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure to afford methyl 4-chloro- 3,5-dinitrobenzoate (9.5 g, yield: 90%) as light yellow solid. LC-MS m/z (M+H):

Ste -2: Synthesis of ethyl l-nitro-10H-phenothiazine-3-carboxylate:

To a stirred solution of methyl 4-chloro-3,5-dinitrobenzoate (1 g, 8mmol, step-1) in ethanol (15 mL) was added 2-aminobenzenethiol (2.08 g, 8 mmol) followed by sodium hydroxide (960 mg, 24 mmol) at 0 °C. Then reaction mixture was stirred at room temperature for 4 h. The progress of the reaction was monitored by TLC. Reaction mixture was filetered and washed with ethanol (10 mL) dried under vacuum to afford ethyl l-nitro-10H-phenothiazine-3-carboxylate (1.3 g, yield: 52%) as brown color solid. LC-MS m/z (M+H): 317.1

Step-3: Synthesis of ethyl l-amino-10H- henothiazine-3-carboxylate:

To a stirred solution of ethyl l-nitro-10H-phenothiazine-3-carboxylate (800 mg, 2.53 mmol, step-2) in 1 ,4-Dioxan/H20 (10:3 mL, 7:3) was added Zn (1.31 g, 20.25 mmol) followed by NH4C1 (1.09 g, 20.25 mmol) at 0 °C. Then reaction mixture was stirred at room temperature for 5 h.The progress of the reaction was monitored by TLC. The reaction mixture was filtered through a pad of celite, washed with ethyl acetate (100 mL). Take filtrate washed with brine solution (1x100 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure to afford ethyl 1 -amino- 10H-phenothiazine-3- carboxylate (600 mg,) as yellow solid. LC-MS m/z (M+H): 286.1

Step-4: Synthesis of ethyl l-(l-(tert-butoxycarbonyl)piperidin-4-ylamino)-10H-phenothia zine- 3-carboxylate:

To a stirred solution of ethyl 1 -amino- 10H-phenothiazine-3-carboxy late (250 mg, 0.70 mmol, step-3,) and tert-butyl 4-oxopiperidine-l-carboxylate (550 mg, 1.92 mmol) in 1 ,2-dichloroethane (50 rriL) was added molecular sieves powder (lOg), stirred at room temperture for lh. Then added sodium tri acetoxy borohydride (4 g, 19.2 mmol) at room temperature, then the reaction mixture was stirred at rt for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was filtered through celite pad washed with DCM (100 mL). Take filtrate concentrated under reduced pressure.. The crude compound was purified by column chromatography (silica gel 60-120 mesh, eluted with 10% EtoAc in DCM). The pure fractions were collected and concentrated under reduced pressure to afford ethyl l -(l-(tert- butoxycarbonyl)piperidin-4-ylamino)-10H-phenothiazine-3-carb oxylate (700 mg, yield: 70%) as yellow solid. LC-MS m/z (M+H): 470.2

Step-5: Synthesis of ethyl l-(piperidin-4-ylamino)-10H-phenothiazine-3-carboxylate:

To a stirred solution of ethyl l-(l-(tert-butoxycarbonyl)piperidin-4-ylamino)-10H- phenothiazine-3-carboxylate (1.8 g, 3.83 mmol, step-4) in DCM (2 mL) was added 1,4- Dioxan.HCl (5mL, 4M) at 0 °C, then the reaction mixture was stirred at rt for lh. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure to obtained crude was basified with saturated NaHCC solution up to pH~7 then extracted with 5%MeoH:DCM (2x100 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure to afford ethyl l-(piperidin- 4-ylamino)-10H-phenothiazine-3-carboxylate (1.2 g, yield: 85%) as yellow solid. LC-MS m/z (M+H): 370.1

Step-6: Synthesis of ethyl l-(l-(2-(tert-butoxycarbonylamino)ethyl)piperidin-4-ylamino) -10H- phenothiazine-3-carboxylate:

To a stirred solution of ethyl l -(piperidin-4-ylamino)-10H-phenothiazine-3-carboxylate (1.2 g, 3.25 mmol, step-5) in acetonitrile (20 mL) was added potassium carbonate (1.34 g, 9.75 mmol) at 0 °C, stirred for 5 min. Then added tert-butyl 2-bromoethylcarbamate (1.09 g, 4.87 mmol) at 0 °C, reaction mixture was stirred at 80 °C for 2h. The progress of the reaction was monitored by TLC. The reaction mixture was cooled to room temperature diluted with water (30 mL) and extracted with 5%MeoH:DCM (2x20 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by column chromatography (silica gel 60-120 mesh, eluted with 2-3% MeoH in DCM). The pure fractions were collected and concentrated under reduced pressure to afford ethyl 1-(1- (2-(tert-butoxycarbonylamino)ethyl)piperidin-4-ylamino)-10H- phenothiazine-3-carboxylate (1.1 g, yield: 66%) as yellow solid. LC-MS m/z (M+H): 513.2

Step-7: Synthesis of 10-tert-butyl 3-ethyl l-(tert-butoxycarbonyl(l-(2-(tert- butoxycarbonylamino)ethyl)piperidin-4-yl)amino)-10H-phenothi azine-3,10-dicarboxylate:

To a stirred solution of ethyl l-(l -(2-(tert-butoxycarbonylamino)ethyl)piperidin-4-ylamino)- 10H-phenothiazine-3-carboxylate (800 mg, 1.56 mmol) in ACN (10 niL) was added DMAP (476 mg, 3.90 mmol) and followed byDi-tert-butyl dicarbonate (1.02 g, 4.68 mmol) at 0 °C,reaction mixture was heated at 80 °C for 3 h. The progress of the reaction was monitored by TLC. Reaction mixture was cooled to room temperature, diluted with water (100 mL) then extracted with ethyl acetate (2x60 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by column chromatography (silica gel 60-120 mesh, eluted with 10% EtoAc in petether). The pure fractions were collected and concentrated under reduced pressure to afford and dried under reduced pressure to afford 10-tert-butyl 3-ethyl l-(tert-butoxycarbonyl(l -(2-(tert- butoxycarbonylamino)ethyl)piperidin-4-yl)amino)-10H-phenothi azine-3,10-dicarboxylate (400 mg, yield: 36%) as grey solid. LC-MS m/z (M+H): 712.1

Step-8: Synthesis of 10-(tert-butoxycarbonyl)-l-(l-(2-(tert- butoxycarbonylamino)ethyl)pipe thiazine-3-carboxylic acid:

To a stirred solution of 10-tert-butyl 3-ethyl l-(tert-butoxycarbonyl(l -(2-(tert- butoxycarbonylamino)ethyl)piperidin-4-yl)amino)-10H-phenothi azine-3,10-dicarboxylate (350 mg, 0.491 mmol) in ethanol (3.5 mL) was added sodium hydroxide (58.9 mg, 1.47 mmol) in H ₂ 0 (0.7 mL) at 0°C, Then the reaction mixture was stirred at room temperature for 16h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure, to obtained crude was acidified with 10% citric acid solution (pH~4) solid was obtained. Filter The solid dried under vacuum to afford 10-tert-butyl 3-ethyl l-(tert- butoxycarbonyl(l -(2-(tert-butoxycarbonylamino)ethyl)piperidin-4-yl)amino)-10 H- phenothiazine-3,10-dicarboxylate (200 mg, yield: 69%) as off white solid. LC-MS m/z (M+H): 685.3

Step-9: Synthesis of tert-butyl l-(l-(2-(tert-butoxycarbonylamino)ethyl)piperidin-4-ylamino) - 3-(pyrrolidine- 1-carbonyl)- 1 OH-phenothiazine- 10-carboxylate:

To a stirred solution of 10-tert-butyl 3 -ethyl l -(tert-butoxycarbonyl(l-(2-(tert- butoxycarbonylamino)ethyl)piperidin-4-yl)amino)-10H-phenothi azine-3,10-dicarboxylate (50 mg, 0.085 mmol) and pyrrolidine (9.1 mg, 0.12 mmol) in DMF (1 mL) was added DIPEA (33.1 mg, 0.25 mmol) at 0°C, stirred for 5min. Then added HATU (48.7 mg, 0.12 mmol) at 0 °C. Then the reaction mixture was stirred at room temperature for 2h. The progress of the reaction was monitored by TLC. The reaction mixture was poured into ice-water (lOmL) stirred for 5min solid is obtained. Filter the solid dried under vacuum. The solid was purified by preparative TLC (eluted with 3% MeOH /DCM) to afford tert-butyl l -(l-(2-(tert-butoxycarbonylamino)ethyl)piperidin-4-ylamino)- 3- (pyrrolidine-l -carbonyl)-lOH-phenothiazine-lO-carboxylate (20 mg, yield: 30%) as off white solid lH NMR (400 MHz, DMSO-d6) δ 1.36 (s, 9H), 1.38 (s, 9H), 1.52-1.61 (m, 1H), 1.72 - 1.92 (m, 4H), 2.05-2.15 (m, 2H), 2.29 - 2.36 (m, 2H), 2.71- 2.89 (m, 2H), 2.97 - 3.07 (m, 2H), 3.30 - 3.33 (m, 2H), 3.37-3.43 (m, 2H), 5.74 (brs, 1H), 6.60 - 6.68 (m, 1H), 6.72 (d, J= 3.12Hz, 2H), 7.25 (t, J = 7.72Hz, 1H), 7.35 (t, J = 7.71Hz, 3H), 7.46 (d, J = 7.70Hz, 1H), 7.73 (d, J = 7.79Hz, 1H). LC-MS m/z (M+H): 638.3

Step-10: Synthesis of (l-(l-(2-aminoethyl)piperidin-4-ylamino)-10H-phenothiazin-3- yl)(pyrrolidin-l-yl)methanone:

To a stirred solution of tert-butyl l -(l-(2-(tert-butoxycarbonylamino)ethyl)piperidin-4-ylamino)- 3- (pyrrolidine-l -carbonyl)-lOH-phenothiazine-lO-carboxylate (20 mg, 0.031mmol) in DCM (1.5 mL) was added 1,4-Dioxan.HCl (lmL, 4M) at 0 °C, then the reaction mixture was stirred at room temperature for 2h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure obtained solid was washed with diethyl ether (2x3 mL) and dried under reduced pressure to afford (l-(l-(2-aminoethyl)piperidin-4-ylamino)-10H- phenothiazin-3-yl)(pyrrolidin-l-yl)methanone (15 mg, yield: 78%) as yellow solid.

1H NMR (400 MHz, OMSO-d ₆ ) δ 1.79 (s, 4H), 1.92 (d, J =11.4Hz, 2H), 2.12 (d, J =12.4Hz, 2H), 3.13 (d, J =10.1Hz, 2H), 3.30 - 3.39 (m, 9H), 3.61 (d, J = 10.2Hz, 4H), 6.40-6.47 (m, 1H), 6.58 (s, 1H), 6.78 (t, J = 6.6Hz, 1H), 6.91-7.02 (m, 3H), 8.11 (s, 1H), 8.31 (s, 3H), 8.59 (s, 1H), 10.94 (brs, 1H)LC-MS m/z (M+H): 438.2

Some examples of compounds synthesised by the method of Scheme XXVIII are provided in Table XXVIII

Figure 23 shows general synthetic scheme XXIX for the synthesis of a selected 1, 3, 8- trisubstituted phenothiazine.N-Acylation of 2-bromo-amino pyridines (XXIXa) followed by neucleophilic substitution reaction with a thiol surrogate yielded compound XXIXd. Deprotection of alkyl chain using NaOEt followed by nucleophilic substitution and Smiles rearrangement give compound XXIXe. Acid-amine coupling or reductive amination of XXIXe followed by deprotection using HC1 yielded the corresponding salts.

Compound 257: 3-amino-N-(8-(trifluoromethyl)-5H-benzo[e]pyrido[3,4-b] [l,4]thiazin-6- yl)cyclohexanecarboxamide

Step 1 :N-(3-bromopyridin-4-yl)acetamide

To a stirred solution of 3 -bromopyridin-4-amine (1.0 g, 5.780 mmol) in dichloromethane (20 mL) was added di isopropyl ethylamine(1.5mL, 8.678mmol) followed by acetyl chloride (0.45mL, 6.345mmol) at 0 °C and starred at room temperature for 12 h. After completion of the reaction mixture was diluted with dichloromethane (20 mL) and washed with saturated sodium bicarbonate solution (20 mL), dried over sodium sulphate, and concentrated. The crude product obtained as N-(3-bromopyridin-4-yl)acetamide as off white solid.(1.24 g, 80%).

1H NMR (400 MHz, OMSO-d ₆ ) δ 3.62 (s, 3H), 6.43 (brs, 2H), 6.77 (d, J = 8.16Hz, 1H), 7.33 (d, J = 2.05Hz, 1H), 7.65 (dd, J= 8.5Hz, 1H)

LC-MS m/z (M+H): 215.0

Step 2: 3-ethylheptyl 3-(4-acetamidopyridin-3-ylthio)propanoate

To a stirred solution of N-(3-bromopyridin-4-yl)acetamide (1.1 g, 0.086 mmol) and 2-ethylhexyl 3-mercaptopropanoate(1.4 mL, 6.138 mmol) in toluene (20 mL) was added DIPEA (4.85 mL 27.62 mmol,) followed by Xanthpos (0.061 g, 0.1023 mmol) at room temperature. Then purged with argonfor 10 min, then added Pd ₂ (dba) 3 (0.046 g, 0.0511 mol), then purged with argonfor 5 min. Reaction mixture was stirred at 1 10 °C for 4 h. The reaction mixture was filtered through celite, concentrated under reduced pressure. Obtained crude product was purified on gradient coloum with 30-40% Ethyl acetate/hexane as eluant to gives 3-ethylheptyl 3-(4- acetamidopyridin-3-ylthio)propanoate as colour less oil (1.4 g, 77% ). 1H NMR (400 MHz, CDC1 ₃ ) δ 0.87-0.95 (m, 6H), 1.25-1.40 (m, 9H), 1.55-1.65 (m, 1H), 2.35 (s, 3H), 2.57 (t, J= 7.02 Hz, 2H), 2.95 (t, J= 7.06 Hz, 2H), 4.07-4.09 (m, 2H),8.43(d, J= 5.5 Hz, 1H), 8.47 (t, J= 5.7 Hz, 1H), 8.68 (s, 1H), 8.98 (brs, 1H). LC-MS m/z (M+H): 353.48

Step 3: sodium 4-acetamidopyridine-3-thiolate

To a stirred solution of 3-ethylheptyl 3-(4-acetamidopyridin-3-ylthio)propanoate (1.5 g, 4.225 mmol) in ethanol (20 mL) was added 21% sodium ethoxide solution in ethanol (3mL) at 0 °C, then stirred at 0 °C for 1 h, evaporated the solvent under reduced pressure, to get sodium 4- acetamidopyridine-3-thiolate (0.75 g) obtained was forwarded to the next step. Step 4: 6-nitro-8-(trifluoromethyl)-5H-benzo[e]pyrido[3,4-b] [l,4]thiazine

A stirred suspension of sodium 4-acetamidopyridine-3-thiolate (0.75g, 3.94 mmol), 2-chloro-l,3- dinitro-5-(trifluoromethyl)benzene(1.2g, 4.33 mmol), in DMF (7.5 mL) was heated at 100 °C for 12 h. To this added 50 mL of ice cold water, extracted the compound into EtOAc, dried the organic layer over sodium sulphate, filtered and concentrated. Obtained crude was purified on gradient coloum with 20-30% EtOAc/hexane as eluant to gives 6-nitro-8-(trifluoromethyl)-5H- benzo[e]pyndo[3,4-b][l,4]thiazine ( 0.8 g, 65 %) , as off Black solid 1H NMR (400 MHz, DMSO-i/ ₆ ) δ 7.13 (d, J = 5.39Hz, 1H), 7.79-7.80 (m, 1H), 8.05(s, 1H), 8.14(s, 1H), 8.18 (d, J = 6.12Hz, 1H), 9.85(brs, 1H) LC-MS m/z (M+H): 314.2 Step 5: 8-(trifluoromethyl)-5H-benzo[e]pyrido[3,4-b] [l,4]thiazin-6-amine

To a stirred solution of 6-nitro-8-(trifluoromethyl)-5H-benzo[e]pyrido[3,4-b][l,4]thi azine ( 0.3 g, 0.961 mmol) in methanol (10 mL) was added a solution of pd/C (10%, water wet, 0.04 g) , and stirred at room temperature for 12h. The reaction mixture was filtered through celite and concentrated to get 8-(trifluoromethyl)-5H-benzo[e]pyrido[3,4-b][l,4]thiazin-6-a mine(0.200 g,74 %))as a brown solid.1H NMR (400 MHz, OMSO-d ₆ ) δ 5.50 (brs, 2H) 6.52 (s, 1H), 6.73 (d, J= 5.39Hz, 1H), 6.77 (d, J= 1.86Hz, 1H), 7.93(s, 1H), 8.03 (d, J= 5.37Hz, 1H), 8.29 (brs, 1H).LC-MS m/z (M+H): 284.1

Step 6: tert-butyl 3-(8-(trifluoromethyl)-5H-benzo[e]pyrido[3,4-b][l,4]thiazin- 6- ylcarbamoyl)cyclohexylcarbamate

To a stirred solution of 3 -((tert-butoxycarbonyl)amino)cyclohexanecarboxylic acid(0.21 g, 0.864 mmol) in pyridine(2 mL) was added a solution POCI ₃ (0.8 mL) at 0 °C and starred at 0 °C for 10 min. and was added a solution of 8-(trifluoromethyl)-5H-benzo[e]pyrido[3,4-b][l,4]thiazin-6- amine in pyridine (2 ml) at 0 °C and starred at 0 °C for lh. After completion of the reaction mixture was poured in to ice, and basified with saturated sodium bi carbonate solution and extracted compound with EtOAc (3 X 25ml), dried the organic layer over sodium sulphate filtered and concentrated. The crude was purified by gradient column chromatography (product eluted with 20 % EtOAc/hexane) to give tert-butyl3-(8-(trifluoromethyl)-5H-benzo[e]pyrido [3,4-b][l,4]thiazin-6-ylcarbamoyl)cyclohexylcarbamate as off white solid (0.120 g, 35%). 1H NMR (400 MHz, DMSO-i/ ₆ ) 51.29-1.33 (m ,4H), 1.35-1.42 (m, 12H), 1.62-1.89 (m, 4H), 2.01- 2.09 (m, 1H), 6.84(d, J = 5.55Hz, 1H), 7.21 (s, 1H), 7.34(s, 1H), 7.99(s, 1H), 8.08 (d, J = 5.39Hz, 1H), 8.49(s, 1H), 9.38(s, 1H).LC-MS m/z (M+H): 509.56

Step 7: 3-amino-N-(8-(trifluoromethyl)-5H-benzo[e]pyrido[3,4-b] [l,4]thiazin-6- yl)cyclohexanecarboxamide

To a stirred solution of tert-butyl3-(8-(trifluoromethyl)-5H-benzo[e]pyrido [3,4-b][l,4]thiazin-6- ylcarbamoyl)cyclohexylcarbamate (0. lg, 0.196 mmol) in dichloromethane (20 mL) was added a solution of HCl/dioxane (4M, 2.0 mL) at 0 °C and starred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure, neutralized with saturated sodium bicarbonate solution, extracted with dichloromethane (30 ml), and concentrated. The crude was purified by preparative TLC(3%MeOH in DCM) to get the 3-amino-N-(8-(trifluoromethyl)-5H- benzo[e]pyrido[3,4-b][l ,4]thiazin-6-yl)cyclohexanecarboxamide (0.04 g, 50%). as off white solid 1H NMR (400 MHz, OMSO-d ₆ ) δ 1.01 -1.07 (m, 1H), 1.22-1.34 (m, 4H), 1.77-1.83 (m, 2H), 1.89 (d, J = 10.3Hz, 1H), 2.05 (d, J = 1 1.7Hz, 1H), 2.52-2.59 (m, 1H), 2.69-2.75 (m, 1H), 6.91 (d, J = 5.3Hz, 1H), 7.19 (s, 1H), 7.43 (s, 1H), 7.98 (s, 1H), 8.07 (d, J = 5.3Hz, 1H)

LC-MS m/z (M+H): 409.1

Some examples of compounds synthesised by the method of Scheme XXIX are provided in Table XXIX

Figure 24 shows general shynthetic scheme XXX for the synthesis of selected substituted phenothiazenes. 2- amino cyclo hexanol of XXXa were protected with boc and and neucleophilic substitution reaction with a thiol surrogate by using potassium thio acetate via., mesylation XXXd. followed bu insituSmiles rearrangement resulted in the formation of trisubstituted phemothiaxzenes XXXd. Nitro group reduction with Zn/NH4C1 gave compound XXXg, Reductive amination of XXXhwith an appropriate ketone resulted in compound XXXi, followed by deprotection resulted compound XXX.

Compound 259: .N-(piperidin-4-yl)-7-(trifluoromethyl)-2,3,4,4a,10,10a-hexa hydro-lH- phenothiazin-9-amine:

Step-1: Synthesis of tert-butyl (2-hydroxycyclohexyl) carbamate:

To a stirred solution of 2-aminocyclohexanol hydrochloride (6.5 g, 43.04 mmol) in DCM (120 mL) was added TEA (15 mL, 107.61 mmol) at 0 °C followed by di tert butyl carbamate (8.75 g, 40.89 mmol) and stirred the reaction mixture at room temperature for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with DCM(100 mL), washed with water (200 mL), saturated NaHCC solution (100 mL). The organic layer was separated, dried over anhydrous sodium sulphate and concentrated under reduced pressure to afford tert-butyl (2- hydroxy cyclohexyl) carbamate (8.5 g, yield: 92%) as off white solid.

Step-2: Synthesis of 2-((tert-butoxycarbonyl)amino)cyclohexyl methanesulfonate:

To a stirred solution of tert-butyl (2-hydroxycyclohexyl) carbamate (8.5 g, 39.53 mmol) in (100 mL) was DCM added tri ethyl amine (17 mL, 118.60 mmol) followed by mesyl chloride (4 mL, 51.39 mmol) at 0 °C. After stirring the reaction mixture for 15 min allowed to stir at room temperature for 3 h. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with DCM (100 mL), washed with brine (100 mL) and saturated NaHC0 ₃ solution (100 mL). The organic layer was separated dried over anhydrous sodium sulphate and concentrated under reduced pressure to afford 2-((tert-butoxycarbonyl)amino)cyclohexyl methanesulfonate (10.1 g, yield: 87%) as off white solid.

Step-3: Synthesis of 2-((tert-butoxycarbonyl)amino)cyclohexyl methanesulfonate:

To a stirred solution of 2-((tert-butoxycarbonyl)amino)cyclohexyl methanesulfonate (10 g, 34.12 mmol,) in DMF (100 mL) was added potassiumthioacetate (11.6, 102.38 mmol) at 0 °C and stirred the reaction mixture at 130 °C for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was poured into ice-cold water (100 mL), extracted with ethyl acetate (3x100 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 4-5% EtOAc in Hexane). The pure fractions were collected and concentrated under reduced pressure to afford 2-((tert-butoxycarbonyl)amino)cyclohexyl methanesulfonate (2.4 g, yield: 26%) as off white solid.

Step-4: Synthesis of tert-butyl (2-mercaptocyclohexyl) carbamate:

To a stirred solution of Lithium aluminium hydride (0.42 g, 10.98 mmol) in di ethyl ether (5 mL) was added 2-((tert-butoxy carbonyl) amino) cyclohexyl methanesulfonate (1 g, 3.66 mmol) in di ethyl ether (35 mL) at 0 °C drop wise over as period of 20 min and stirred the reaction mixture at room temperature for 1 h. The progress of the reaction was monitored by TLC. The reaction mixture was cooled to 0 °C, added 2N NaOH solution (2.5 mL) and stirred for 10 min; solid formed was filtered through a pad of celite, washed with ethyl acetate (2x25 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated (20-25 °C) under reduced pressure to afford tert-butyl (2-mercaptocyclohexyl)carbamate (380 mg, crude) as colorless solid. The crude compound was used in the next step without any purification.

Step-5: Synthesis of 2-((tert-butoxycar nyl)amino)cyclohexyl methanesulfonate:

To a stirred solution of tert-butyl (2-mercaptocyclohexyl)carbamate (700 mg, 3.03 mmol), 2-chloro- l,3-dinitro-5-(trifluoromethyl)benzene (818 mg, 2.42 mmol) in EtOH (10 mL) was added NaOH (364 mg, 9.09 mmol) at 0 °C and stirred the reaction mixture at room temperature for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with water (30 mL), extracted with ethyl acetate (3x25 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 4-5% EtOAc in Hexane). The pure fractions were collected and concentrated under reduced pressure to afford 2-((tert-butoxycarbonyl) amino)cyclohexyl methanesulfonate (180 mg, yield: 14%) as yellow oil.LC-MS m/z (M+H): 418.43

Step-6: Synthesis of tert-butyl 9-amino-7-(trifluoromethyl)-2,3,4,4a-tetrahydro-lH- phenothiazine-10(10aH)-carboxylate:

To a stirred solution of 2-((tert-butoxycarbonyl)amino)cyclohexyl methanesulfonate (35 mg, 0.08 mmol) in MeOH (1 mL), H ₂ 0 (1 mL) was added NH ₄ C1 (22 mg, 0.41 mmol) followed by Zn dust (27 mg, 0.41 mmol) at 0 °C and stirred the reaction mixture at room temperature for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was filtered through a pad of celite, washed with ethyl acetate (2x10 mL). The filtrate was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with EtOAc in Hexane) followed by preparative TLC ( to afford tert-butyl 9-amino-7-(trifluoromethyl)-2,3,4,4a-tetrahydro-lH-phenothia zine-10(10aH)- carboxylate (25 mg, yield: 80%) as yellow gummy liquid.LC-MS m/z (M+H): 389.33

To a stirred solution of tert-butyl 9-amino-7-(trifluoromethyl)-2,3,4,4a-tetrahydro-lH- phenothiazine-10(1 OaH)-carboxylate (60 mg, 0.15 mmol) in DCM (2 niL) was added 4M HC1 in Dioxane (2 mL) at 0 °C and stirred the reaction mixture at room temperature for 2 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The residue was basified by using aqueous saturated NaHCC , extracted with ethyl acetate (2x20 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by preparative HPLC (20% EtOAc in Hexane) to afford 7-(trifluoromethyl)-2,3 ,4,4a, 10,10a-hexahydro-lH- phenothiazin-9-amine (35 mg, yield: 79%) as colorless liquid. H NMR (400 MHz, DMSO-d6) δ 1.30-1.44(m, 2H), 1.55-1.68 (m, 4H), 1.70-1.81(m, 1H), 1.90 -1.99 (m, 1H), 3.26-3.31 (m, 1H), 3.73-3.74 (m, 1H), 5.03 (s, 1H), 5.15(s, 1H), 6.48 (s, 1H), 6.58 (d, J = 1.99 Hz, 1H). LC-MS m/z (M+H): 289.3

Step-8: Synthesis of tert-butyl 4-((7-(trifluoromethyl)-2,3,4,4a,10,10a-hexahydro-lH- phenothiazin-9-yl)amino) piperidine-l-carboxylate

12

To a stirred solution of 7-(trifluoromethyl)-2,3,4,4a,10,10a-hexahydro-lH-phenothiazi n-9-amine (30 mg, 0.10 mmol) in DCE (3 mL) was added molecular sieves (200 mg), tert-butyl 4- oxopiperidine-1 -carboxylate (24 mg, 0.12 mmol), Acetic acid (0.01 mL) at 0 °C. After stirring the reaction mixture for 1 h, was added sodium triacetoxy borohydride (177 mg, 0.83 mmol) and stirred the reaction mixture at room temperature for 48 h. The progress of the reaction was monitored by TLC. The reaction mixture was filtered through a pad of celite, washed with DCM (2x10 mL). The combined organic layer was washed with saturated NaHCC (25 mL). The aq layer was extracted with DCM (2x10 mL), dried over anhydrous sodium sulphate and concentrated under reduced pressure to afford tert-butyl 4-((7-(trifluoromethyl)-2,3,4,4a,10,10a-hexahydro-lH-phenoth iazin-9- yl)amino) piped dine- 1 -carboxylate (60 mg, crude) as brown color liquid. The crude compound was used in the next step without any purification.

Step-9: Synthesis of N-(piperidin-4-yl)-7-(trifluoromethyl)-2,3,4,4a,10,10a-hexah ydro-lH- phenothiazin-9-amine:

To a stirred solution of tert-butyl 4-((7-(trifluoromethyl)-2,3 ,4,4a, 10,1 Oa-hexahydro-lH- phenothiazin-9-yl) amino) piped dine- 1 -carboxylate (50 mg, 0.10 mmol) in DCM (2 mL) was added 4M HC1 in 1 ,4-dioxane (2 mL) at 0 °C and stirred the reaction mixture at room temperature for 1 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The residue was diluted with ethyl acetate (20 mL), washed with saturated NaHCC (20 mL).The organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by preparative TLC (3% MeOH in DCM) to afford N-(piperidin-4-yl)-7-(trifluoromethyl)-2,3 ,4,4a, 10,10a-hexahydro-lH- phenothiazin-9-amine (9 mg, yield: 24%) as grey solid. ^X H NMR (400 MHz, OMSO-d ₆ ) δ 8.05 (s, 1H), 6.56 (s, 1H), 6.48 (s, 1H), 5.43 (s, 1H), 4.92 (d, J = 6.8Hz, 1H), 3.74 (s, 1H), 3.54-3.51 (m, 1H), 3.26-3.23 (m, 5H), 2.93 (t, J = 1 1.6Hz, 2H), 2.03-1.90 (m, 2H), 1.79-1.78 (m, 1H), 1.65-1.51 (m, 5H), 1.49-1.33 (m, 2H) LC-MS m/z (M+H): 372.1

Figure 25 shows general reaction scheme XXXI for the synthesis of selected tert-butyl (2- aminoethyl) carbamate alkylated with ethyl 3-chloropropanoate in the presence of base to gave XXXIb, which were further protected with boc anhydride to give N-protected ester XXXIc, XXXIc was ester reduction with lithium aluminum hydride to gave XXXId and followed by oxidation with martin's reagent to give corresponding aldehydes XXXIe. Reductive amination of compound XXXI e with various aldehydes or ketones yielded corresponding ^-alkylated phenothiazines XXXIf, which were further deprotected to give the corresponding free amines XXXIg. And alkylated to give ΧΧΧΠι, further protection of the ΧΧΧΠι with boc anhydride to give tetra boc protected compounds of XXXIi. Further Buchwald coupling of compound XXXIi with various amines, followed by deprotection gave XXXI 1 with corresponding salts. And Further Suzuki coupling of compound of XXXIi with boronic acids followed by deprotection gave XXXI 3 with corresponding salts and also with XXXIj double bond reduction with platinum oxide and followed by deprotection gave XXXI 2

Compound 346: Nl-(2-aminoethyl)-N3-(7-(3, 5-dimethylpiperidin-l-yl)-3-(trifluoromethyl)- lOH-phenothiazin-l-yl) propane-1, 3-diamine hydrochloride

Step 1: Ethyl 3-((2-((tert-butoxycarbonyl) amino) ethyl) amino) propanoate:

To a solution of tert-butyl (2-aminoethyl) carbamate (20 g, 124.843 mmol) in acetonitrile (200 mL) was added potassium carbonate (68.9 g, 499.275 mmol) and ethyl 3- chloro formate (18.9 g, 138.38 mmol) at room temperature. Reaction mixture was stirred at room temperature for 16 h. Progress of the reaction was followed by TLC (5 % methanol/DCM). After completion of the reaction, diluted with water (400 mL) and extracted with ethyl acetate (150 mL x 3). Organic layer was dried over sodium sulphate, filtered and concentrated to give crude product. This crude product purified by gradient column chromatography (product eluted with 100% DCM) to give ethyl 3-((2-((tert- butoxycarbonyl) amino) ethyl) amino) propanoate as colorless liquid (20 g, 61 %). ¹ H NMR (400 MHz, DMSO-de) 51.1 (t, J = 7.16 Hz, 3H), 1.4 (s, 9H), 1.6 (br s, 1H), 2.3(t, J= 5.08 Hz, 2H), 2.4- 2.5 (m, 2H), 2.7 (t, J= 6.8 Hz, 2H), 2.9-3.0 (m ,2H),4.0(q, J= 7.04 Hz 2H).

Step 2: Ethyl 3-((tert-butoxycarbonyl)(2-((tert- butoxycarbonyl)amino)ethyl)amino)propanoate:

To a solution of ethyl 3-((2-((tert-butoxycarbonyl) amino) ethyl) amino) propanoate (20 g, 76.79 mmol) in DCM (200 mL) was added triethyl amine (31.1 g, 307.342 mmol) and followed by di tert butyl di carbonate (28.7 g, 131,651 mmol) at 0 °C. Reaction mixture was stirred at room temperature for 16 h. After completion of the reaction, diluted with DCM (200 mL) and washed with water (100 mL x 2). Organic layer was dried over sodium sulphate, filtered and evaporated to give crude product. This crude product purified by gradient column chromatography (product eluted with 10 % ethyl acetate/n-hexane) to give ethyl 3-((tert-butoxycarbonyl)(2-((tert- butoxycarbonyl)amino)ethyl)amino)propanoate as colorless liquid (23 g, 83 %). ^X H NMR (400 MHz, DMSO-de) 51.1 (t, J = 7.08 Hz, 3H), 1.4 (s, 19H), 2.4-2.5 (m, 1H), 2.9-3.0 (m, 2H), 3.1 (t, J = 6.36 Hz, 2H), 3.2 (s, 1H), 3.3 (t , J= 7.08 Hz, 2H), 4.0-4. l(m, 2H), 6.8(br s, 1H).

Step 3: Tert-butyl (2-((tert-butoxycarbonyl) amino) ethyl)(3-hydroxypropyl)carbamate:

H

HO ^' ^ ^/ ^ N ^N " boc

I

boc

To a solution of ethyl 3-((tert-butoxycarbonyl)(2-((tert- butoxycarbonyl)amino)ethyl)amino)propanoate (12 g, 33.291 mmol) in THF (120 mL) was added lithium aluminum hydride (1.53 g, 40.326 mmol) at 0 °C. Reaction mixture stirred at room temperature at room temperature for 3 h. After completion of the reaction, quenched with ice water and diluted with ethyl acetate, filtered through celite for remove inorganic salts. Organic layer was separated and dried over sodium sulphate, filtered and evaporated to give crude product. This crude product purified by gradient column chromatography (product eluted with 40 % ethyl acetate/n-hexane) to give tert-butyl (2-((tert-butoxycarbonyl) amino) ethyl)(3- hydroxypropyl)carbamate (8 g, 75.5 %). 1H NMR (400 MHz, DMSO-d ₆ ) 51.4 (s, 18H), 1.5 (br s, 2H), 3.0-3. l(m, 2H), 3.1-3.2 (m, 2H), 3.3 (q, J= 5.88 Hz, 2H), 4.4 (br s, 1H), 6.8 (br s, 1H).

Step 4: Tert-butyl (2-((tert-butoxycarbonyl) amino) ethyl) (3-oxopropyl) carbamate:

boc

To a solution of tert-butyl (2-((tert-butoxycarbonyl) amino) ethyl)(3-hydroxypropyl)carbamate (8 g, 25.157 mmol) in DCM (160 mL) was added Dessmartin's reagent (16 g, 37.735 mmol) at 0 °C. Reaction mixture stirred at room temperature at room temperature for 3 h. After completion of the reaction, diluted with DCM (100 mL) and filtered through celite, filtrate was washed with water (100 mL). Organic layer was separated and dried over sodium sulphate, filtered and evaporated to give crude residue, which was triturated with diethyl ether (100 mL) and filtered to remove the inorganic salts. Filtrate was concentrated to give crude product, which was used for the next step without further purification (8 g, crude). Step 5: tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl) amino) propyl) amino) ethyl) carbamate

To a stirred solution of 7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l -amine (5.2 g, 14.448 mmol) in dichloroethane (100 mL) was added tert-butyl (2-((tert-butoxycarbonyl) amino) ethyl) (3-oxopropyl) carbamate (5.485 g, 17.337 mmol) and 4A° Molecular sieves powder (10 g) at room temperature After stirring the reaction mixture for 1 h, was added sodium triacetoxy boro hydride (9.184 g, 43.344 mmol). Reaction mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with saturated NaHCC solution, extracted with DCM (2x100 mL). The combined organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 20% EtOAc in Hexane) to afford tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)- lOH-phenothiazin-l-yl) amino) propyl) amino) ethyl) carbamate as Off white solid (3 g, 31.4%). 1H NMR (400 MHz, DMSO-d ₆ ) 51.3-1.5 (m, 11H), 1.7-1.8 (m, 2H), 3.0-3. l (m, 4H), 3.2(t, J= 6.4 Hz, 2H), 3.3 (t, J= 9.64 Hz, 2H), 6.4 (s, 1H), 6.5(s, 1H), 6.8(d, J = 8.7 Hz, 2H), 7.2(m, 2H), 8.0 (s, 1H). Step 6: tert-butyl 7-bromo-l-((tert-butoxycarbonyl)(3-((tert-butoxycarbonyl)(2- ((tert- butoxycarbonyl)amino)ethyl)amino)propyl)amino)-3-(trifluorom ethyl)-10H-phenothiazine- 10-carboxylate

To a stirred solution of tert-butyl (2-((3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-yl) amino) propyl) amino) ethyl) carbamate (3 g, 4.534 mmol) in Acetonitrile (50 mL) was added DMAP (1.9 g, 15.871 mmol) followed by ditert-butyl dicarbonate (5 g, 22.673 mmol) at 0 °C and stirred the reaction mixture at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, reaction mixture was diluted with ethyl acetate (100 mL) and washed with ethyl acetate (50 mL). The combined organic layer was dried over anhydrous sodium sulpahte filtered and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 15% EtOAc in Hexane) to afford tert-butyl 7-bromo-l-((tert-butoxycarbonyl) (3-((tert-butoxycarbonyl) (2-((tert- butoxycarbonyl)amino) ethyl)amino ) propyl) amino)-3 -(trifluoromethyl)- 10H-phenothiazine- 10-carboxylate as off white solid (2.1 g, 53.8%). 1H NMR (400 MHz, DMSO-d ₆ ) ) 51.2 (s, 5H), 1.3-1.4 (m, 12H), 1.6-1.8 (m, 2H), 3.1-3.3 (m, 5H), 3.4 (s, 1H), 3.6 (s,2H), 6.2 (br, 1H), 6.7- 6.8(m, 1H), 6.9(m, 1H), 7.5 (d, J = 7.68 Hz, lH) 7.7 (d, J = 8.48 Hz, 1H), 7.7 (s, 1H)

Step 7: tert-butyl l-((tert-butoxycarbonyl)(3-((tert-butoxycarbonyl)(2-((tert- butoxycarbonyl)amino)ethyl)amino)propyl)amino)-7-(3,5-dimeth ylpiperidin-l-yl)-3- (trifluoromethyl)- 1 OH-phenothiazine- 10-carboxylate

To a stirred solution of tert-butyl 7-bromo-l-((tert-butoxycarbonyl) (3-((tert-butoxycarbonyl) (2- ((tert-butoxycarbonyl)amino) ethyl)amino ) propyl) amino)-3 -(trifluoromethyl)- 10H- phenothiazine-10-carboxylate (120 mg, 0.139 mmol), 3,5-dimethylpiperidine (31.5 mg,0.278 mmol) in 1 4 Dioxane(3 mL), water (1 mL) was added sodium hydroxide (13.92 mg, 0.348 mmol) at room temperature. After degassed with argon for 10 min was added Pd ₂ (dba)3 (12.75 mg, 0.013 mmol), tert-butylX-phos ( 8.85 mg, 0.020 mmol) again degassed for 5 min and stirred the reaction mixture at 110 °C for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with water (10 mL), extracted with ethyl acetate (2x20 mL). The combined organic layer was dried over anhydrous sodium sulpahte and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 25% ethyl acetate/n-Hexane) to afford tert-butyl l-((tert-butoxycarbonyl)(3-((tert- butoxycarbonyl)(2-((tert-butoxycarbonyl)amino)ethyl)amino)pr opyl)amino)-7-(3,5- dimethylpiperidin-l-yl)-3-(trifluoromethyl)-10H-phenothiazin e-10-carboxylate as off white solid (30 mg, 24.19%). 1H NMR (400 MHz, DMSO-d ₆ ) δ 0.6-0.8(m, 1H), 0.9 (d, J = 2.6 Hz ,6H), 1.2 (s, 4H), 1.3 (s, 9H), 1.4 (s, 18H), 1.5-1.6 (m, 5H), 1.7-1.9 (m, 1H), 3.6-3.7 (m, 4H), 5.4 (br, 1H), 6.7-6.8 (m, 2H), 6.9 (s, 1H), 7.2 (d, J= 8.68 Hz, 1H), 7.3 (d, J= 2.08 Hz, 1H). Step 8: Nl-(2-aminoethyl)-N3-(7-(3,5-dimethylpiperidin-l-yl)-3-(trif luoromethyl)-10H- phenothiazin-l-yl)propane-l,3-diamine hydrochloride

To a stirred solution of tert-butyl l-((tert-butoxycarbonyl)(3-((tert-butoxycarbonyl)(2-((tert- butoxycarbonyl)amino)ethyl)amino)propyl)amino)-7-(3,5-dimeth ylpiperidin-l-yl)-3- (trifluoromethyl)-lOH-phenothiazine-lO-carboxylate (30 mg, 0.003 mmol)in DCM (1 mL) was added 4M HC1 in 1,4-dioxane (2 mL) at 0 °C and stirred the reaction mixture at room temperature for 2 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was washed with n-pentane (2x2 mL) and dried under reduced pressure to afford Nl -(2-aminoethyl)-N3-(7-(3,5-dimethylpiperidin-l-yl)-3- (trifluoromethyl)-10H-phenothiazin-l -yl)propane-l,3-diamine hydrochloride as grey solid (16 mg, 90.3%) ^X H NMR (400 MHz, DMSO-d ₆ ) δ 0.8-0.9 (m, 7H), 1.1 -1.3 (m, 1H), 1.7-1.8 (m, 1H), 2.0-2.1 (m ,2H), 2.2-2.3 (m, 2H), 3.0-3.2 (m, 9H), 3.3-3.4 (m, 4H), 6.5(d, J = 6 Hz 2H), 7.2-7.5 (m, 8.3(br s, 3H), 9.3 (br s, 2H). MS m/z (M+H): 494.23

Compound 358: Nl-(2-aminoethyl)-N3-(7-(cyclopent-l-en-l-yl)-3-(trifluorome thyl)-10H- phenothiazin-l-yl) propane-1, 3-diaminehydrochloride

Step 9: tert-butyl

l((3((tertbutoxycarbonyl)(2((tertbutoxycarbonyl)amino)eth yl)amino)propyl)amino)-7- (cyclopent-l-en-l-yl)-3-(trifluoromethyl)-10H-phenothiazine- 10-carboxylate:

To a stirred solution (200 mg, 0.232 mmol, compound-1) in 1 4 Dioxane/H ₂ 0 (8 mL) was added potassium carbonate (76 mg, 0.696 mmol) at room temperature. After degassed with argon for 10 min was added boronic acid (52 mg, 0.464 mmol) and finally added Pd2(dppf)C12 DCM complex. (10 mg, 0.116 mmol) again degassed for 5 min and stirred the reaction mixture at 110 °C for 12 h. Progress of the reaction was monitored by TLC. The reaction mixture was filtered through a pad of celite; the filtrate was concentrated under reduced pressure. The crude compound was purified by combi-flash chromatography (eluted with 2-3% EA in Pet Ether) followed by preparative TLC to afford tert- butyll((3((tertbutoxycarbonyl)(2((tertbutoxycarbonyl)amino)e thyl)amino)propyl)amino)-7-

(cyclopent-l-en-l-yl)-3-(trifluoromethyl)-10H-phenothiazi ne-10-carboxylate as off white solid (82 mg, yield: 47%). 1H NMR (400 MHz, DMSO-d6) δ 1.2 (s, 18H), 1.31 -1.35 (m, 4H), 1.4 (s, 18H), 1.7-1.8 (m,2H), 1.91 -1.92 (m, 2H), 2.6 (d, J = 4.5Hz, 2H), 3.19-3.25 (m, 4H), 3.29-3.33 (m, 1H), 3.61 -3.69 (m, 2H), 6.11 (brs, 1H), 6.33(s, 1H), 6.72 (s, 1H), 6.94 (s, 1H), 7.45(d, J = 16.4Hz, 1H), 7.5 (s, 1H), 7.64 (d, J = 8.2Hz, 1H).

LC-MS m/z (M+H): 749.2

Step 10: Nl-(2-aminoethyl)-N3-(7-(cyclopent-l-en-l-yl)-3-(trifluorome thyl)-10H- phenothiazin-l-yl) propane-1 3-diamine hydrochloride

To a stirred solution of tert- butyll((3((tertbutoxycarbonyl)(2((tertbutoxycarbonyl)amino)e thyl)amino)propyl)amino)-7- (cyclopent-l-en-l-yl)-3-(trifluoromethyl)-10H-phenothiazine- 10-carboxylate (25 mg, 0.0334 mmol) in DCM (1 mL) was added 1,4-dioxane HC1 (2 mL) at 0 °C and stirred the reaction mixture at room temperature for 2 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was triturated with n- pentane (2x2 mL) and dried to afford Nl-(2-aminoethyl)-N3-(7-(cyclopent-l -en-l-yl)-3- (trifluoromethyl)-lOH-phenothiazin-l -yl) propane-1, 3-diamine hydrochloride as pale yellow solid (12 mg, yield: 85%). ¾ NMR (400 MHz, DMSO-d ₆ ) 51.8- 1.9 (m, 2H), 2.1-2.19 (m, 2H), 2.4-2.5 (m, 2H), 2.5-2.55 (m, 1H), 3.19-3.25 (m, 10H), 3.5-3.7 (m, 2H), 6.14 (s, 1H), 6.5 (d, J = 6.8Hz, 2H), 7.1 (s, 3H), 8.23 (br s, 3H), 8.4 (br s, 1H), 9.2 (br s, 2H). LC-MS m/z (M+H): 449.3

Compound 353: Nl-(2-aminoethyl)-N3-(7-cyclopentyl-3-(trifluoromethyl)-10H- phenothiazin- 1-yl) propane-1, 3-diamine h drochloride

Step 11: tert-butyl l-((3-((tert-butoxycarbonyl)(2-((tert- butoxycarbonyl)amino)ethyl)amino)propyl)amino)-7-cyclopentyl -3-(trifluoromethyl)-10H- phenothiazine-10-carboxylate

To a stirred solution of tert- butyll((3((tertbutoxycarbonyl)(2((tertbutoxycarbonyl)amino)e thyl)amino)propyl)amino)-7- (cyclopent-l-en-l-yl)-3-(trifluoromethyl)-10H-phenothiazine- 10-carboxylate (60 mg, 0.0802 mmol) in Toluene (5 mL) was added platinum oxide (30 mg) at room temperature under hydrogen atmosphere for 6 h. The progress of the reaction was monitored by TLC. The reaction mixture was filtered through celite bed filtrate was concentrated under reduced pressure, crude product was purified by prep TLC (20 % ethyl acetate/ n-Hexane) to afford tert-butyl l-((3-((tert- butoxycarbonyl)(2-((tert-butoxycarbonyl)amino)ethyl)amino)pr opyl)amino)-7-cyclopentyl-3- (trifluoromethyl)-lOH-phenothiazine-lO-carboxylate as colorless gel (42 mg, yield: 70%). LC-MS m/z (M+H): 751.2

Step 12: Nl-(2-aminoethyl)-N3-(7-cyclopentyl-3-(trifluoromethyl)-10H- phenothiazin-l-yl) propane-1, 3-diamine hydrochloride

To a stirred solution of tert-butyl l -((3-((tert-butoxycarbonyl)(2-((tert- butoxycarbonyl)amino)ethyl)amino)propyl)amino)-7-cyclopentyl -3-(trifluoromethyl)-10H- phenothiazine-10-carboxylate (42 mg, 0.056 mmol) in DCM(1 mL) was added 1,4-dioxane HC1 (2 mL) at 0 °C and stirred the reaction mixture at room temperature for 3 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was triturated with n-pentane (2x2 mL) and dried to afford Nl -(2- aminoethyl)-N3-(7-cyclopentyl-3-(trifluoromethyl)-10H-phenot hiazin-l-yl) propane-1, 3-diamine hydrochloride as pale yellow solid (24 mg, yield: 96%). 1H NMR (400 MHz, DMSO-d ₆ ) δ 1.4-1.5 (m, 2H), 1.5-1.6 (m, 2H), 1.7(m, 2H), 1.9-2.0(m, 2H), 2.01(t, J = 6.6Hz, 2H), 2.71-2.81 (m, 1H), 3.1-3.2 (m, 8H), 6.54 (d, J = 7.6Hz, 2H), 6.8 (s, 1H), 6.9 (d, J = 12.2Hz, 1H), 7.03 (s, 1H), 8.2-8.3 (m, 4H), 9.2 (br s, 2H).

Some examples of compounds synthesized by the method of Scheme XXXI are provided in Table XXXI

Table XXXI

Figure 26 shows general reaction scheme XXXII for the synthesis of selectedl, 3, 6- trisubstituted phenothiazines. Tert butyl acryl ate was treated with protected 2- amine ethanol to gave XXXIIa, further reduction with lithium aluminum hydride and followed by oxidation with dessmartin' reagent to give corresponding aldehydes (XXXIId). Reductive amination of compound XXXIId with XXIIId yielded corresponding n-alkylated phenothiazines XXXIIe and further deprotection gave XXXII with corresponding salts. Compound 343: N-(3-(2-aminoethoxy) propyl)-7-bromo-3-(trifluoromethyl)-10H- phenothiazin-l-amine hydrochloride

Step-1: tert-butyl 3-(2-(tert-butoxycarbonylamino)ethoxy)propanoate:

To a stirred solution of tert-butyl acrylate (16 g, 124.223 mmol), in 1 , 4-dioxane (100 mL) was added 60% KOH in water at room temperature and stirred the reaction mixture at room temperature for 24 h. The progress of the reaction was monitored by TLC. Reaction mixture diluted with ethyl acetate (100 mL), washed with water (100 mL). The organic layer was dried over anhydrous sodium sulphate and concentrated under reduce pressure. The crude compound was purified by gradient column chromatography (eluted with 0-2 MeOH% in DCM) to afford tert-butyl 3-(2-(tert- butoxycarbonylamino)ethoxy)propanoate (12 g, yield: 66%) as colourless oil H NMR (400 MHz, DMSO-i/ ₆ ) δ, 1.40-1.44 (s, 9H), 1.45-1.50 (s, 9H), 2.45-2.49 (m, 2H), 3.29-3.33 (m, 2H), 3.49-3.52 (m, 2H), 3.67-3.70 (m, 2H), 4.95 (br s, 1H)

Step-2: tert-butyl 2-(3-hydroxypropoxy) ethyl carbamate: To a stirred solution of tert-butyl 3-(2-(tert-butoxycarbonylamino)ethoxy)propanoate (5 g, 17.301 mmol) in dry THF (100 mL) was added lithium aluminium hydride at 0 °C and stirred the reaction mixture at room temperature for 4h. The progress of the reaction was monitored by TLC. Reaction mixture was quenched with aqueous saturated sodium sulphate solution at 0 °C and stirred the reaction mixture at room temperature for 10 min, diluted with ethyl acetate (50 mL). The reaction mixture was filtered through a pad of celite and washed with ethyl acetate (20 mL). The filtrate was dried over anhydrous sodium sulphate and concentrated under reduce pressure. The crude compound was purified by gradient column chromatography (eluted with 30-50% in ethyl acetate/ n-Hexane) to afford tert-butyl 2-(3- hydroxypropoxy) ethylcarbamate (2.5 g, yield: 67%) as colorless oil H NMR (400 MHz, DMSO-<¾) δ, 1.39 (s, 9H), 1.57-1.62 (m, 2H), 3.02-3.37 (m, 2H), 3.38-3.45 (m, 3H), 4.33 (m, 1H), 6.78 (brs, 1H)

Step-3: tert-butyl 2-(3-oxopropoxy) ethylcarbamate:

H

0 ^{<i s} ^ ¾'^ -^ "Boc

To a stirred solution of tert-butyl 2-(3 -hydroxypropoxy) ethylcarbamate (600 mg, 2.575 mmol) in DCM (10 mL) was added Desmartin's reagent (1.64 g, 3.86 mmol) at 0 °C and stirred the reaction mixture at room temperature for 4 h. The progress of the reaction was monitored by TLC. The reaction mixture was filtered through a pad of celite; the filtrate was extracted with ethyl acetate (2x20 mL). The combined organic layer was washed with water (15 mL), dried over anhydrous sodium sulphate and concentrated under reduce pressure to afford tert-butyl 2-(3-oxopropoxy) ethylcarbamate as colour less oil (340 mg, crude). The crude product was used for next step without further purification. Step-4: tert-butyl 2-(3-(7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-ylamino ) propoxy)ethylcarbamate:

To a stirred solution of 7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l -amine (420 mg, 1.169 mmol) and tert-butyl 2-(3-oxopropoxy)ethylcarbamate (235 mg, 1.169 mmol), molecular sieves (3 g) in DCE (10 mL) was stirred at room temperature for lh. And was added sodium triacetoxyborohydride at 0 °C under nitrogen atmosphere and stirred the reaction mixture at room temperature for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with saturated sodium bicarbonate solution (20 mL), extracted with DCM (3x30 mL). The combined organic layer was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 20% ethyl acetate/n-Hexane) to afford tert-butyl 2-(3-(7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l - ylamino)propoxy)ethylcarbamate (120 mg, yield: 18%) as brown oil. 1H NMR (400 MHz, OMSO-d ₆ ) δ 1.30 (s, 9H) 1.83 (t, 2H), 3.07 (d, J = 5.85Hz, 2H), 3.12 (d, J = 5.85Hz, 2H), 3.31 -3.37 (m, 3H), 3.52 (m, 2H), 6.5407 (d, J = 5.39 Hz, 2H), 6.76-6.80 (m, 2H), 7.18-7.20 (m, 2H), 8.05 (s, 1H). LC-MS m/z (M+H): 561.9

Step-5: Synthesis of N-(3-(2-aminoethoxy)propyl)-7-bromo-3-(trifluoromethyl)-10H- phenothiazin- 1-amine hydrochloride:

To a stirred solution of tert tert-butyl 2-(3-(7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l- ylamino)propoxy)ethylcarbamate (120 mg, 0.213 mmol) in 1 ,4-Dioxane (3 mL) was added 4M HCI in 1,4-Dioxane (3 mL) at 0 °C, and stirred the reaction mixture at room temperature for 2 h.

The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was washed with diethyl ether (2x3 mL), pentane (3 mL) and dried under reduced pressure to afford N-(3-(2-aminoethoxy)propyl)-7-bromo-3- (trifluoromethyl)-lOH-phenothiazin-l -amine hydrochloride (90 mg, 84%) as pale green solid. Ή NMR (400 MHz, DMSO-i/ ₆ ) δ 1.80-1.92 (m, 2H) 2.90 -3.11 (m, 2H), 3.15-3.22 (m, 2H), 3.50- 3.62 (m, 4H) 6.51 (s, 1H), 7.13 (d, 2H), 7.93 (br s, 3H), 8.73 (s, 1H). LC-MS m/z (M+H): 462 Figure 27 shows general reaction scheme XXXIII for the synthesis of selectedl, 3, 6- trisubstituted phenothiazines. 3 amino cyclo hexyl carboxylic was protected boc anhydride to gave XXXIIIa, and followed by acid amide coupling with weinreb amine to giave XXXnic further reduction with lithium aluminum hydride to give corresponding aldehydes (XXXIIId).

Reductive amination of compound XXXIIId with XXIIId yielded corresponding n-alkylated phenothiazines XXXDIe and further deprotection gave XXXIII with corresponding salts.

Compound 343: N-((3-aminocyclohexyl)methyl)-7-bromo-3-(trifluoromethyl)-10 H- phenothiazin-l-amine hydrochloride

Step-1: 3-(tert-butoxycarbonylamino) cyclohexane carboxylic acid

To a stirred solution of 3 -aminocyclohexanecarboxylic acid (5 g, 34.92 mmol), in 1 , 4-dioxane (50 mL) was added sodium hydroxide and stirred the reaction mixture at room temperature for 24h. The progress of the reaction was monitored by TLC. Reaction mixture was cooled to 0°C then added IN HCl to adjust the P ^H to 4,the solid precipitates were filtered and washed with water (100 mL) dried under vacuum to give 3-(tert-butoxycarbonylamino) cyclohexane carboxylic acid (7.0 g, 82%) as white coloursolid . H

NMR (400 MHz, OMSO-d ₆ ) δ 0.985-1.3(m, 4H), 1.39 (s, 9H), 1.6-1.8 (m, 3H), 1.9-2.0 (m, 1H), 2.2- 2.4(m, 1H), 3.15-3.3 (m, 1H), 6.73-6.75(d, J = 8.0 Hz, 1H), 12.04 (s, 1H). LC- MS m/z (M+H): 144.05 Step-2: tert-buty!3-(methoxy (methyl) carbamoyl) cyclohexylcarbamate

To a stirred solution 3-(tert-butoxycarbonylamino)cyclohexane carboxylic acid (3.0 g, 12.341 mmol) in dry DCM (30 mL) was added TEA at 0 °C and stirred for 15 min then added Ν,Ο- dimethylhydroxylamine and Py-Bop then stirred at room temperature for 14 h. The progress of the reaction was monitored by TLC. Reaction mixture was quenched with saturated NaHCC solution at 0 °C and stirred the reaction mixture at room temperature for 10 min, Organic layer was separated and dried over anhydrous sodium sulphate and concentrated under reduce pressure. The crude compound was purified by combi flash (eluted with 20 ethyl acetate/n-Hexane) to afford the title compound (3.2 g, yield: 91%) as brown color oil H NMR (400 MHz, DMSO-i/ ₆ ) δ, 1.1 -1.3 (m,4H) ,1.39 (s, 9H), 1.57- 1.62 (m, 1H), 1.7 (brs, 3H), 2.62 - 2.81 (m, 1H), 3.02 (s, 3H), 3.258-3.298 (m, 1H), 3.653 (s, 3H), 5.742 (s, lH),6.736-6.75 (d, J =7.6 Hz 1H), LC-MS m/z (M+H): 187.1

Step-3: tert-butyl 3-formylcyclohexylcarbamate

To a stirred solution of tert-butyl3-(methoxy (methyl) carbamoyl) cyclohexylcarbamate (3.5 g, 12.195 mmol) in THF (10 mL) was added lithium aluminum hydride (463 mg, 12.195 mmol) at 0°C and stirred the reaction mixture at room temperature for 0.5 h. The progress of the reaction was monitored by TLC. The reaction mixture was quenched with saturated Na ₂ S0 ₄ then filtered the reaction mass, filtrate was dried over anhydrous sodium sulphate then concentrated under vacuum to get title compound tert-butyl 3-formylcyclohexylcarbamate (1.3 g, crude) as colorless oil, which was used for the next step without further purification. 1H NMR (400 MHz, OMSO-d ₆ ) δ, 0.9-1.29 (m, 9H), 1.39 (s, 9H), 1.57-1.9(m, 5H), 1.97-2.00 (m, 2H), 4.0(m, 1H), 4.33(m, 1H), 6.77-6.79 (d, J =8.0 Hz 1H), 9.59 (s. lH).

Step-4: tert-butyl 3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l-ylamino) methyl) cyclohexylcarbamate

To a stirred solution of tert-butyl 3-formylcyclohexylcarbamate (1.062 g, 4.678 mmol) in DCE (10 mL) added 7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l -amine (1.4 g,3.899 mmol) molecular sieves (3 g) was stirred at room temperature for lh, then added sodium triacetoxyborohydride at 0 °C under nitrogen atmosphere and stirred the reaction mixture at same temperature for 16 h. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with sat NaHCC solution (20 mL), extracted with DCM(3x30 mL). The combined organic layer was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by gradient column chromatography (eluted with 20% ethyl acetate/n-Hexane) to afford tert-butyl 3-((7-bromo-3-(trifluoromethyl)-10H-phenothiazin-l -ylamino) methyl) cyclohexylcarbamate (2.2 g, 22%) as brown oil. ¾ NMR (400 MHz, OMSO-d ₆ ) δ 1.33 (s, 9H), 1.751 - 1.957 (m, 5H), 1.977-2.02 (m, 1H), 2.50 (br s, 2H), 3.37 (s, 1H), 5.4 (s, 1H), 6.530 - 6.558 (d, J = 1 1 Hz, 2H), 6.75-6.822 (m, 2H), 7.16-7.19 (m, 2H), 8.125 (s, 1H). LC-MS m/z (M+H): 572.1. Step-5: N-((3-aminocyclohexyl)methyl)-7-bromo-3-(trifluoromethyl)-10 H-phenothiazin-l-amine hydrochloride

To a stirred solution of tert-butyl 3-((7-bromo-3-(trifluoromethyl)-l OH-phenothiazin-1 - ylamino)methyl)cyclohexylcarbamate (70 mg, 0.225 mmol) in 1 ,4-Dioxane (3 mL) was added 4M HC1 in dioxane (3 mL) at 0 °C, and stirred the reaction mixture at room temperature for 2 h. The progress of the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure. The crude compound was washed with diethyl ether (2x3 mL), pentane(3 mL) and dried under reduced pressure to afford N-((3-aminocyclohexyl)methyl)-7-bromo-3-(trifluoromethyl)-10 H-phenothiazin-l - amine mg, yield: 82%) as pale yellow solid. 1H NMR (400 MHz, OMSO-d ₆ ) δ 0.842- 0.866 (m, 1H) 0.981 -1.014 (m, 1H), 1.199-1.308 (m, 3H), 1.761 -1.828 (m, 1H) 6.49-6.51 (d, J =l l Hz 2H), 7.07-7.09 (m, 3H), 7.901 (s, 3H), 8.660 (s, 1H). LC-MS m/z (M+H): 471.98

Characterisation of the Synthesised Compounds

Table 13 below provides LC-MS data on the compounds synthesised and indicates which general synthetic method (Scheme number) was used to obtain the compound.

Table 13

LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

Nl -(3-

(trifluoromethy 1)- 1 OH- Ill

97 C , phenothiazin-1 - 339.38 340.1

yl)propane-l ,3 -diamine

1 -(2-aminoethy l)-3 -(3 - (trifluoromethy 1)- 1 OH- III

98 phenothiazin-1 - 384.07 385.1

yl)thiourea

(R)-N-(3- o (trifluoromethy 1)- 1 OH- III

100 phenothiazin-1 - 393.11 394.1

yl)piperidine-3- carboxamide

3 -(piperazin-1 -yl)-N- (3 -(trifluoromethy 1)- III

101 COX ° ^" 1 OH-phenothiazin-1 - 422.47 423.1

yl)propanamide

3-amino-N-(3- (trifluoromethy 1)- 1 OH- III

105 phenothiazin-1 - 407.13 408.1

yl)cyclohexanecarboxa

mide LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - Ill

106 yl)piperidine-3- 393.11 394.1

carboxamide

N-((4- aminocyclohexyl)meth III

108 yl)-3 -(trifluoromethyl)- 393.15 394.2

1 OH-phenothiazin-1 - amine

N,N-dimethyl-3-(3- phenyl-10H- IV

110 phenothiazin- 10- 360.17 361.5

yl)propan- 1 -amine

3-(3-(lH-indol-2-yl)- 1 OH-phenothiazin-10- IV

111 yl)-N,N- 399.18 400.2

dimethylpropan-1 - amine

2-amino-N-(3-

O NH ₂ (trifluoromethy 1)- 1 OH- III

113 phenothiazin-1 - 407.13 408.1

yl)cyclohexanecarboxa

mide LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

2-(4,4-difluoro-l- hydroxycyclohexyl)-N- Ill

115 (3 -(trifluoromethy 1)- 458.11 459.1

O _s X _CF3 1 OH-phenothiazin-1 - yl)acetamide

3 -(3 -cyclopropyl- 10H- phenothiazin- 10 -y 1) - IV

116 N,N-dimethylpropan- 324.17 325.2

1 -amine

3-(3-(lH- benzo [d] imidazol-5 - IV yl)-l OH-phenothiazin-

117 10-yl)-N,N- 400.17 401.2

dimethylpropan-1 - amine

(lR,3S)-3-amino-N-(3- (trifluoromethy 1)- 1 OH- III

118 phenothiazin-1 - 407.13 408.1

yl)cyclohexanecarboxa

mide

3-amino-N-(3-cyano- 1 OH-phenothiazin-1 - III

119 364.14 365.2

O _s XX _CN yl)cyclohexanecarboxa

mide LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

2-(4,4- difluorocyclohexyl)-N- Ill

132 (3 -(trifluoromethy 1)- 442.11 443.4

1 OH-phenothiazin-1 - yl)acetamide

N-((3- aminocyclohexyl)meth III

133 yl)-3 -(trifluoromethyl)- 393.15 394.2

1 OH-phenothiazin-1 - amine

3-amino-N-(3- (trifluoromethy 1)- 1 OH- III

134 phenothiazin-1 - 401.08 402.1

yl)benzamide

3-(7-(lH-indol-2-yl)- 10H- benzo[b]pyrido[2,3- VI

135 e][l,4]thiazin-10-yl)- 400.17 401.2

N,N-dimethylpropan- 1 -amine

3-(3-(l -(2- aminoethyl)-lH-indol- IV NH ₂ 2-yl)-10H-

136 rr ^N n phenothiazin- 10 -y 1) - 442.22 443.2

N,N-dimethylpropan- 1 -amine LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

3-(3-(lH- benzo[d]imidazol-2- VII yl)-l OH-phenothiazin-

137 10-yl)-N,N- 400.17 401.2

dimethylpropan-1 - amine

2-amino-N-(3- (trifluoromethy 1)- 10H- III

139 phenothiazin-1- 408.12 408.1

yl)piperidine-4- carboxamide

N-(l -(2- aminoethyl)piperidin- III

140 4-yl)-3- 408.16 409.2

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine r^NH N-(piperidin-4-yl)-3-

141 (trifluoromethy 1)- 10H- III

365.12 366.1

phenothiazin- 1 -amine

3-(3-(benzo[d]thiazol- 2-yl)-10H- VII

143 phenothiazin- 10 -y 1) - 419.15 418.2

N,N-dimethylpropan- 1 -amine LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

3-(3-(5-methoxy-lH- indol-2-yl)-10H- IV

144 phenothiazin- 10 -y 1) - 429.19 430.2

N,N-dimethylpropan-

1 -amine

3 -hydroxy -N-(3 - (trifluoromethy 1)- 10H- III

148 phenothiazin-1- 408.11 409.0

yl)cyclohexanecarboxa

mide

3 -(aminomethy l)-N-(3 - (trifluoromethy 1)- 10H- III

149 phenothiazin-1- 421.14 422.2

yl)cyclohexanecarboxa

mide

3-(3-(lH-indol-2-yl)-

IV

150 n ^N n « 1 OH-phenothiazin-10- 371.15 372.1

yl)propan- 1 -amine

2-(10-(3-

(dimethylamino)propyl IV ° )-l 0H-phenothiazin-3 -

151 yl)-lH-indole-5- 538.17 425.2

carbonitrile 2,2,2- trifluoroacetate salt LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

2-(10-(3-

(dimethylamino)propyl IV

152 )-l 0H-phenothiazin-3 - 415.17 416.2

yl)-lH-indol-5-ol

3-(3-(2- methoxypyridin-4-yl)- IV 1 OH-phenothiazin-10-

153 yl)-N,N- 391.53 392.2

dimethylpropan-1 - amine l -(2-aminoethyl)-N-(3- (trifluoromethy 1)- 1 OH- III ^^ phenothiazin-1 -

155 ¾xx _CFj v„: - yl)piperidine-4- 550.15 437.2

carboxamide 2,2,2- trifluoroacetate

3-amino-N-(3- (trifluoromethy 1)- 10H- III

156 phenoxazin-1- 391.15 392.2

yl)cyclohexanecarboxa

mide LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

Γ NH N-(pyrrolidin-3 -yl)-3 -

174 (trifluoromethy 1)- 10H- 351.10 352.1 Ill phenothiazin- 1 -amine

3-amino-N-(3- (trifluoromethy 1)- 10H- III

175 phenothiazin-1- 379.10 380.1

yl)cyclobutanecarboxa

mide

3-chloro-N2-(3- (trifluoromethy 1)- 10H- III

178 phenothiazin-1- 373.06 374.1

yl)propane-l ,2-diamine

N-((4-(2- aminoethyl)cyclohexyl III

179 )methyl)-3- 421.18 423.0

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine

Nl -(3-

(trifluoromethy 1)- 10H- III

180 phenothiazin-1- 379.13 380.0

yl)cyclohexane- 1,4- diamine LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

3 -(3 -(5 -(aminomethyl)- lH-indol-2-yl)-10H- IV

181 cc Ac phenothiazin- 10 -y 1) - 428.20 429.2

N,N-dimethylpropan- 1 -amine

N-(((lR,3S)-3- aminocyclohexyl)meth III

182 yl)-3 -(trifluoromethyl)- 393.15 394.0

1 OH-phenothiazin-1 - amine r^NH 3 -chloro-N-(piperidin-

184 4-yl)-10H- 331.09 332.1 III

C phenothiazin- 1 -amine

3-(3-(lH-indol-2-yl)- 1 OH-phenothiazin-10- IV

186 Cf l ^« yl)-N-methylpropan-l - 385.16 386.2

amine

N-(((l S,3S)-3- aminocyclohexyl)meth III H ⁰ ^ ^CFs yl)-3 -(trifluoromethyl)-

T

187 O'

1 OH-phenothiazin-1 - 507.14 394.0

amine 2,2,2- trifluoroacetate

LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l -(2- aminoethyl)piperidin- Ill

214 4-yl)-3-chloro-10H- 374.13 375.0

phenothiazin- 1 -amine l -((l -(2- aminoethyl)piperidin- III

215 4-yl)amino)-10H- 365.17 366.3

phenothiazine-3 - carbonitrile

Nl -(2-aminoethyl)-Nl - (2-(4-((3- III

(trifluoromethy 1)- 10H- phenothiazin-1-

216 yl)amino)piperidin-l - 608.24 495.2

yl)ethyl)ethane-l ,2- diamine 2,2,2- trifluoroacetate salt

9-((l -(2- aminoethyl)piperidin- 4-yl)amino)-7- XVII

217 x i (trifluoromethy 1)- 10H- 433.15 434.2

phenothiazine-3 - carbonitrile LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l -(2- aminoethyl)piperidin- Ill

218 4-yl)-10-methyl-3- 422.18 423.2

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine

Nl -(2-(4-((3- (trifluoromethy 1)- 10H- III phenothiazin-1-

219 yl)amino)piperidin-l - 451.20 451.2

yl)ethyl)ethane-l ,2- diamine

N-((l-(2- aminoethyl)piperidin- III

220 3-yl)methyl)-3- 422.18 423.2

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine y NH

N-(azepan-4-yl)-3 - fuoromethy III

221 (tril 1)- 10H- 379.13 380.1

phenothiazin- 1 -amine

3-(3-(l -(2- aminoethyl)-lH-indol- 3c 2-yl)-10H- IV

222 phenothiazin- 10- 642.17 415.2

yl)propan- 1 -amine

trifluoroacetate) salt LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l -(2- aminoethyl)piperidin-

I

4 -y 1) -7 -(aminomethy 1)- XVIII

227 3 -(trifluoromethyl)- 665.58 436.4

1 OH-phenothiazin-1 - amine 2,2,2- trifluoroacetate

N-(l -(2-

N aminoethyl)pyrrolidin- III

228 3-yl)-3- 394.47 395.2

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine

1 -(piperidin-4-yl)-4- ΧΠΙ (trifluoromethyl)imida

232 zo[4,5,l- 375.42 376.1

kl]phenothiazine

Nl -(2-aminoethyl)-Nl - (2-(4-((3-

(trifluoromethy 1)- 1 OH- III

234 phenothiazin-1 - 494.63 495.2

yl)amino)piperidin-l - yl)ethyl)ethane-l ,2- diamine

LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

7-(lH-indol-2-yl)-N- (piperidin-4-yl)-3- vni

243 (trifluoromethy 1)- 10H- 480.56 481.1

phenothiazin- 1 -amine

3-(3-(lH-indol-2-yl)-7- (piperazin-1 -yl)-l 0H-

244 phenothiazin- 10- 569.65 456.2 XX yl)propan- 1 -amine

2,2,2-trifluoroacetate

3-(3-(lH-indol-2-yl)-7- (trifluoromethy 1)- 10H- IV

245 phenothiazin- 10- 439.51 440.1

yl)propan- 1 -amine methyl 9-((l -(2- aminoethyl)piperidin- 4-yl)amino)-7- XXII

247 (trifluoromethy 1)- 10H- 466.53 467.2

phenothiazine-3 - carboxylate

N-(l -(2- aminoethyl)piperidin- III

252 4-yl)-l - 408.49 409.4

H

(trifluoromethy 1)- 10H- phenothiazin-3 -amine LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

3-amino-N-(8-

(trifluoromethyl)-5H- benzo[b]pyrido[4,3-

257 e][l,4]thiazin-6- 408.12 409.1 XXIX yl)cyclohexane- 1 - carboxamide ethyl 10-(3- aminopropy l)-7-( 1 H-

258 indol-2-yl)-10H- 443.17 444.2 IV

phenothiazine-3 - carboxylate

^NH N-(piperidin-4-yl)-7- (trifluoromethyl)-

259 d _{s CF3} 2,3 ,4,4a, 10,10a- 371.16 372.1 XXX hexahydro-lH- phenothiazin-9-amine J 10-(3 -aminopropy l)-7- (lH-indol-2-yl)-10H-

262 phenothiazine-3 - 529.13 416.2 IV

carboxylic acid

trifluoro acetic acid LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

3-amino-N-(7- (trifluoromethy 1)- 10H- benzo[b]pyrido[2,3-

263 e][l,4]thiazin-9- 408.12 409.1 XXIX yl)cyclohexane- 1 - carboxamide

^NH N-(piperidin-4-yl)-7- (trifluoromethy 1)- 10H-

H

264 benzo[b]pyrido[2,3- 366.11 367.1 XXIX e] [ 1 ,4] thiazin-9-amine

3-(3-(piperazin-l-yl)- J 7 -(trifluoromethy 1)-

265 1 OH-phenothiazin-10- 480.11 409.1 IV

yl)propan- 1 -amine

hydrochloride

3-(3-(4-(2- aminoethyl)piperazin- J l -yl)-7-

266 n ^N i (trifluoromethy 1)- 10H- 523.16 452.1 IV

phenothiazin- 10- yl)propan- 1 -amine

hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

3-(8-(lH-indol-2-yl)- 5H- J

benzo[b]pyrido[4,3-

267 e][l,4]thiazin-5- 486.13 374.1 VI yl)propan- 1 -amine

trifluoro acetic acid

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-

268 (trifluoromethy 1)- 10H- 409.15 410.1 XXIX benzo[b]pyrido[2,3- e] [ 1 ,4] thiazin-9-amine

7-(4-(2- aminoethyl)piperazin- 1 -yl)-N-(piperidin-4-

269 yl)-3 -(trifluoromethyl)- 492.23 493.2 XXIII

1 OH-phenothiazin-1 - amine hydrochloride

7-(piperazin-l -yl)-N- (piperidin-4-yl)-3-

270 (trifluoromethy 1)- 10H- 557.12 450.1 XXIII phenothiazin- 1 -amine

hydrochloride

LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z) l -((l -(2- aminoethyl)piperidin- 1 J 4-yl)amino)-N'-(4-

280 fluorophenyl)- 10H- 564.16 493.2 XXVII o ^H phenothiazine-3 - carbohydrazide

hydrochloride l -((l -(2- aminoethyl)piperidin- 1 J

4-yl)amino)-N'-phenyl-

281 10H-phenothiazine-3 - 546.17 475.2 XXVII o ^H

carbohydrazide

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-bromo-3-

282 (trifluoromethy 1)- 10H- 558.02 488.1 XXIII

phenothiazin- 1 -amine

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(3,5-

283 dimethylpiperazin-1 - 664.17 521.2 XXIII

. JJ

yl)-3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l -(2- aminoethyl)piperidin- 4 -y 1) -7 -(pyrr olidin- 1 -

284 yl)-3 -(trifluoromethyl)- 549.17 478.2 XXIII

1 OH-phenothiazin-1 - amine hydrochloride ethyl 9-((l -(2- aminoethyl)piperidin-

4-yl)amino)-7-

(trifluoromethy 1)- 10H-

285 benzo[b]pyrido[2,3- 589.11 482.1 XXIX

e][l,4]thiazine-3- carboxylate

hydrochloride

3-(3-(4-(2- aminoethyl)piperazin- l -yl)-7-(3,5-

286 dimethylpiperidin-1 - 638.23 495.4 XV yl)-l OH-phenothiazin- 10-yl)propan- 1 -amine

hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z) l -((l -(2- aminoethyl)piperidin- 1 J

4-yl)amino)-10H-

287 455.13 384.2

phenothiazine-3 - XXVII carboxamide

hydrochloride

(l -((l -(2- aminoethyl)piperidin- 1 J 4-yl)amino)-10H-

288 phenothiazin-3-

O _g O _y N 509.18 438.2 XXVII yl)(pyrrolidin-l - yl)methanone

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(piperidin-l -

289 563.19 490.2 XXIII yl)-3 -(trifluoromethyl)-

1 OH-phenothiazin-1 - amine hydrochloride

N-(8- azabicy clo[3.2.1 ] octan-

290 3-yl)-3- c X. 391.13 392.2 XXIII

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

3-(3-(4-(2- aminoethyl)piperazin- 1 -yl)-7-(pyrrolidin-l -

291 x ^N xx yl)-l OH-phenothiazin- 560.2 451.3 XV

10-yl)propan- 1 -amine

hydrochloride

7-(4-(2- aminoethyl)piperazin- l -yl)-10-(3-

292 XX ^N XX aminopropy 1)- 10H- 636.2 409.3 XV phenothiazine-3 - carbonitrile trifluoro

acetic acid

(9-((l-(2- aminoethyl)piperidin-

4-yl)amino)-7-

(trifluoromethy 1)- 10H-

293 phenothiazin-3- 577.17 506.2 XXVIII

yl)(pyrrolidin-l- yl)methanone

hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

3-amino-N-(7- (pyrrolidin- 1 -yl)-3 - (trifluoromethy 1)- 1 OH-

294 phenothiazin-1 - 512.16 477.2 XXV yl)cyclohexane- 1 - carboxamide

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(4-

(dimethylamino)piperi

295 . din-l-yl)-3- 606.23 535.2 XXIII

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(azetidin-l-yl)-

296 3 -(trifluoromethyl)- 499.18 502.2 XXIII

1 OH-phenothiazin-1 - amine hydrochloride

Nl -(2-aminoethyl)-N3- (3 -(trifluoromethy 1)-

J

297 1 OH-phenothiazin-1 - 418.12 383.22 XXVI

COX yl)propane-l,3-diamine

hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

Nl -(2-aminoethyl)-N3- (7-bromo-3- (trifluoromethy 1)- 10H-

298 phenothiazin-1- 496.03 461.20 XXVI

yl)propane-l ,3 -diamine

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(3-

(dimethylamino)pyrroli

299 din-l-yl)-3- 592.21 521.22 XXIII

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine

hydrochloride

N-((3- aminocyclohexyl)meth

"fY " ² yl)-7-(pyrrolidin- 1 -yl)-

300 rx ^N f 3 -(trifluoromethyl)- 498.18 463.1 XXV

1 OH-phenothiazin-1 - amine hydrochloride

LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(4-

307 xx ^N xi methylpiperidin-1 -yl)- 577.20 506.3 XXIII

3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

3 -((2- aminoethyl)amino)-N- (7-(3 -methylpiperidin- l -yl)-3-

308 xx ^N xi ^{H hci} (trifluoromethy 1)- 1 OH- 565.17 494.2 XXVII phenothiazin-1 - yl)propanamide

hydrochloride

2-(l-(2- aminoethyl)piperidin- 4-yl)-8-(pyrrolidin-l- yl)-4-

309 575.15 504.43 XXIII

(trifluoromethyl)imida

zo[4,5,l- kl]phenothiazin-l (2H)- one hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(indolin-l -yl)-

310 3 -(trifluoromethyl)- 597.17 526.2 XXIII

1 OH-phenothiazin-1 - amine hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(3-

311 rx x ^HCI methylpyrrolidin-1 -yl)- 563.19 492.22 XXIII

3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

3 -amino-N-(7-bromo- 3 -(trifluoromethyl)- 1 OH-phenothiazin-1 -

312 yl)cyclohexane- 1 - 556.99 488.0 XXV

carboxamide

hydrochloride

3-amino-N-(7-(3- methylpiperidin-1 -yl)- o

3 -(trifluoromethyl)-

313 oi;; 1 OH-phenothiazin-1 - 576.17 505.2 XXV yl)cyclohexane- 1 - carboxamide

hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z) l -((2- aminoethyl)amino)-3 - ((7-(pyrrolidin-l-yl)-3-

317 (trifluoromethy 1)- 1 OH- 573.11 468.22 XXVII phenothiazin-1 - yl)amino)propan-2-one

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(3-

318 fluoropiperidin-1 -yl)- 545.20 510.23 XXIII

3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

7-bromo-N-(l -(2- (piperazin-1 - yl)ethyl)piperidin-4-

319 yl)-3 -(trifluoromethyl)- 591.10 556.17 XXIII

1 OH-phenothiazin-1 - amine hydrochloride

LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(3-

320 ΎχΆ isopropylpiperidin- 1 - 641.21 534.28 XXIII yl)-3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(3-

321 methoxypyrrolidin- 1 - 579.18 508.21 XXIII yl)-3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(2-

322 azaspiro [4.4 ]nonan-2- 603.22 557.22 XXIII yl)-3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z) l -((2-

(dimethylamino)ethyl)

amino)-3-((7- (pyrrolidin- 1 -yl)-3 -

323 (trifluoromethy 1)- 1 OH- 565.15 494.21 XXVII phenothiazin-1 - yl)amino)propan-2-one

hydrochloride

Nl -(7-(pyrrolidin-l - yl)-3 -(trifluoromethyl)-

324 1 OH-phenothiazin-1 - 444.14 409.12 XXIII yl)propane-l,3-diamine

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(2-

325 azaspiro [3.3 ]heptan-2- 539.21 504.2 XXIII l~~p < ^s ^ yl)-3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(3-

326 methoxypiperidin-1 - 557.22 422.2 XXIII yl)-3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(2- mt azabicyclo[2.2. l]hepta

327 n-2-yl)-3- 539.21 504.2 XXIII

(trifluoromethy 1)- 10H- phenothiazin- 1 -amine

hydrochloride

N-(7-(pyrrolidin-l -yl)- 3 -(trifluoromethyl)-

328 1 OH-phenothiazin-1 - 429.09 394.2 XXVII yl)acetamide

hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-bromo-3-

329 methyl-lOH- 468.08 433.15 XXIII phenothiazin- 1 -amine

hydrochloride

LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

Nl -(7-bromo-3- (trifluoromethy 1)- 1 OH-

334 phenothiazin-1 - 487.18 452.2 XXIII

1 yl)propane-l ,3 -diamine

hydrochloride

Nl -(2-aminoethyl)-Nl - (2-(4-((7-(pyrrohdin-l- yl)-3 -(trifluoromethyl)-

335 1 OH-phenothiazin-1 - 635.26 564.3 XXIV yl)amino)piperidin-l - yl)ethyl)ethane-l ,2- diamine hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(2,3-

336 dimethylpyrrolidin-1 - 541.23 506.31 XXIII yl)-3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

N-(l -(2- aminoethyl)piperidin- 4-yl)-7-(2,4-

337 dimethylazetidin- 1 -yl)- 527.17 491.97 XXIII

3 -(trifluoromethyl)- 1 OH-phenothiazin-1 - amine hydrochloride

hydrochloride

hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

Nl-(7-(2- azaspiro [3.3] heptan-2 -y 1) -3 - (trifluoromethyl)- 10H-

347 phenothiazin-l-yl)-N3-(2- 513.19 476.23 XXXI cp aminoethyl)propane-l,3 - diamine hydrochloride

Nl-(3-((2- aminoethyl)amino)propyl)- N7 -ethy 1-N7 -methyl-3 -

348 (trifluoromethyl)- 10H- 511.16 440.18 XXXI

J phenothiazine- 1 ,7-diamine

hydrochloride

Nl-(2-aminoethyl)-N3-(7- (2,4-dimethylpyrrolidin-l - yl)-3 -(trifluoromethyl)-

349 1 OH-phenothiazin- 1 - 515.21 480.24 XXXI y l)propane- 1 ,3 -diamine

hydrochloride

Nl-(7-(2- azabicyclo[2.2. l]heptan-2- yl)-3 -(trifluoromethyl)-

350 1 OH-phenothiazin- 1 -yl)- 513.19 478.23 XXXI

G N3 -(2-aminoethyl)propane- 1,3 -diamine hydrochloride

Nl-(2-aminoethyl)-N3-(7- (hexahydrocyclopenta[b]py

rrol-l(2H)-yl)-3-

351 (trifluoromethyl)- 1 OH- 527.21 492.22 XXXI phenothiazin- 1 -y l)propane- 1,3 -diamine hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l-(2- aminoethyl)piperidin-4-yl)- 8-bromo-3-

352 (trifluoromethyl)- 10H- 522.05 487.2 XXIII phenothiazin- 1 -amine

hydrochloride

Nl-(2-aminoethyl)-N3-(7- cyclopentyl-3-

353 (trifluoromethyl)- 10H- 486.18 451.2 XXXI phenothiazin- 1 -y l)propane- 1,3 -diamine hydrochloride

N-(l-(2- aminoethyl)piperidin-4-yl)- 7-cyclopentyl-3-

354 (trifluoromethyl)- 10H- 548.16 477.2 XXIII phenothiazin- 1 -amine

hydrochloride

N 1 -(2 -aminoethy 1)-N3 -(8 - bromo-3-(trifluoromethyl)-

355 1 OH-phenothiazin- 1 - 496.03 461.1 XXXI y l)propane- 1 ,3 -diamine

hydrochloride

N-(l-(2- aminoethyl)piperidin-4-yl)- 8 -(pyrrolidin- 1 -y 1) -3 -

356 (trifluoromethyl)- 1 OH- 513.19 478.24 XXIII phenothiazin- 1 -amine

hydrochloride LC-MS Synthesis

Cmpd Exact

Structure Name [M+H]+

# mass (Scheme no.)

(m/z)

N-(l-(2- aminoethyl)piperidin-4-yl)- 8 -(3 , 5 -dimethy lpiperidin- 1 -

357 yl)-3 -(trifluoromethyl)- 555.24 520.2 XXIII

1 OH-phenothiazin- 1 -amine

hydrochloride

Nl-(2-aminoethyl)-N3-(7- (cy clopent- 1 -en- 1 -yl) -3 -

358 (trifluoromethyl)- 1 OH- 484.17 449.3 XXXI phenothiazin- 1 -y l)propane- 1,3 -diamine hydrochloride

N-(l-(2- aminoethyl)piperidin-4-yl)- 7 -(cyclopent- 1 -en- 1 -yl) -3 -

359 (trifluoromethyl)- 1 OH- 510.18 475.2 XXXI phenothiazin- 1 -amine

hydrochloride

Table 14 provides a summary of NMR data for the compounds synthesised.

Table 14

Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 2.49-2.56 (m, 4H), 2.66-2.72 (m, 3H), 2.82 (s, 4H),

5.65 (bs, 1H), 6.85-6.90 (m, 2H), 6.98-7.00

101 (d, J= 6.8 Hz, 1H), 7.04-7.07 (t, J= 6.8 Hz,

1H), 7.16 (s, 1H), 7.34 (s, 1H), 8.03 (s, 1H),

9.66 (s, 1H)

¾ NMR (DMSO-de, 400 MHz) 61.18- 1.22 (m, 4H), 1.75-1.78 (m, 2H), 1.84-1.86 (m, 1H),1.98- 2.07(d, J= 12.4 Hz ,1H), 2.59 (s, 1

105 H), 3.45-3.48(m, 1H), 6.83-6.91 (m, 2H),

6.96 (d,J= 6.8 Hz ,1H), 7.02 (t, J= 7.6 Hz, 1H), 7.12 (s, 1H), 8.13 (s, 1H), 7.38 (s, 1H), 8.03 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.47 (s, 1H), 1.62 (s, 2H), 1.97 (d, J= 12 Ηζ,ΙΗ), 2.66 (t, J= 11.2 Hz 1H), 2.80 (t, J= ΙΟ Ηζ ,Ι

106 H), 2.92(d, J= 10 Hz, 1H), 3.17(d, J= 12 Hz,

1H),6.84- 6.90 (m, 2H), 6.98 (d, J= 7.6 Hz ,1H), 7.02 (t, , J = 7.6 Hz ,1H), 7.14 (s, 1H), 7.37 (s, 1H), 8.02 (s, 1H), 9.45 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.43(d, J = 9.6 Hz, 3H), 1.41-1.49 (m, 5H), 1.68 (d, J = 5.6 Hz,3H), 2.65 (s, 1H), 3.01 (t, J= 6 Hz ,2

108 H), 6.54(d, J= 8Hz, 2H), 6.84(t, J= 7.2 Hz,

1H), 6.90 (d, J= 8 Hz, 1H), 6.96 (d, J= 7.2 Hz, 1H), 7.05 (t, J= 7.2 Hz ,1H), 8.01 (s, 1H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.83 (t, J = 8.0 Hz, 2H), 2.16 (s, 6H), 2.40 (s, 2H), 3.93 (t, J= 7.2 Hz, 2H), 6.94 (t, J= 7.2 Hz, 1H),

110 o:;:a _c 7.04 (d, J= 8.0 Hz, 1H), 7.09 (d, J= 8.0 Hz,

1H), 7.15-7.22 (m, 2H), 7.30 (t, J = 7.6 Hz, 1H), 7.39-7.43 (m, 3H), 7.49 (d, J= 8.0 Hz, 1H), 7.61 (d, J= 7.6 Hz, 2H)

^X H NMR (DMSO-de _, 400 MHz) δ 1.94 (s, 6H), 2.31 (s, 2H), 2.66 (s, 2H), 3.96 (s, 2H), 6.80 (s, 1H), 6.96-6.97 (m, 2H), 7.04-7.08

111 (m, 2H), 7.12 (d, J = 8.0 Hz, 1H), 7.19-7.24

(m, 2H), 7.35 (d, J = 8.0 Hz, 1H), 7.48 (d, J = 8.0 Hz, 1H), 7.66-7.69 (m, 2H), 11.40 9s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.22-1.24 (m, 1H), 1.31 -1.35 (m, 1H),1.50-1.53 (m,

113 1H),1.60 (s, 3H), 1.73-1.81 (m, 2H), 2.48- 2.57 (m, 1H),3.35 (s, 1H), 4.66 (bs, 2H),6.80-

6.86 (m, 2H), 6.93-6.98 (m, 2H), 7.13-7.14 (d, J= 5.2 Hz, 1H), 8.8 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.70-1.75 (m, 2H), 1.83-1.97 (m, 4H), 1.97-2.15 (m, 2H), 2.53 (s, 2H), 5.15 (s, 1H), 6.82-6.87 (m,

115 2H), 6.98 (d, J= 7.6 Hz, 1H), 7.05 (t, J= 7.6

Hz, 1H), 7.18 (s, 1H), 7.26 (s, 1H), 7.89 (s, 1H, 9.39 (s, 1H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 0.58 (d, J = 4.0 Hz, 2H), 0.85 (d, J = 6.0 Hz, 2H), 1.73- 1.81 (m, 3H), 2.09 (s, 6H), 2.30 (t, J= 6.0

116 Hz, 2H), 3.84 (t, J = 6.8 Hz, 2H), 6.83 (s,

1H), 6.86-6.91 (m, 3H), 6.97 (d, J= 8.0 Hz,

1H), 7.11 (d, J= 8.0 Hz, 1H), 7.16 (t, J= 8.0 Hz, 1H)

^X H NMR (DMSO-de _, 400 MHz) δ 1.81-1.83 (m, 2H), 2.11 (s, 6H), 2.33 (d, J = 7.6 Hz, 2H), 3.90-3.92 (m, 2H), 6.94 (t, J= 8.0 Hz,

117 1H), 7.04 (d, J= 8.0 Hz, 1H), 7.09 (d, J= 8.0

Hz, 1H), 7.15-7.22 (m, 2H), 7.40-7.55 (m, 3H), 7.65-7.68 (m, 2H), 8.20 (s, 1H), 12.44 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 0.97-1.00 (m, 2H), 1.18-1.25 (m, 2H), 1.29-1.38 (m, 3H), 1.78(d, J = 10.4 Hz, 2H), 1.86 (d, J=7.6 Hz, 1H),2.01 (d, J= 12 Hz, 1H), 2.59 (d, J =

118 11.2 Hz, 1H), 6.87 (d, J= 7.2 Hz, 1H), 6.91

(d, J= 8 Hz, 1H), 6.98 (d, J= 7.2 Hz, 1H), 7.07 (t, J= 7.6 Hz, 1H),7.12 (S,1H); 7.38 (s,lH), 8.03(s,lH)

¾ NMR (DMSO-de _, 400 MHz) δ 1.33-1.37 (m, 2H), 1.49-1.61 (m, 1H), 1.61-1.74 (m, 1H), 1.77-1.99 (m, 4H), 2.31-2.57 (m, 1H),

119 6.84-6.89 (m, 2H), 6.95-6.97 (d, J= 7.2 Hz ,

1H), 7.02-7.06 (t, J= 7.2 Hz, 1H), 7.24 (s, 1H), 7.43 (s, 1H), Ί.6Ί-Ί.6Ζ (m, 1H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ, 0.90-0.96 (m,2H),1.39 ( q, J=12.4 Hz ,2H), 1.85 (d, J=12.0 Hz ,2H),1.95 (d, J=l 1.6 Hz, 2H),

120 2.38-6.2.49 (m,4 H), 6.83-6.91 (m, 1H), 6.97

(d, J= 7.6 Hz, 1H),7.05 (t,J= 7.2

Hz,lH),7.12 (s,lH),7.39 (s,lH), 8.00 (bs,lH)

¾ NMR (DMSO-de _, 400 MHz) δ 1.81-1.83 (m, 2H), 2.10 (s, 6H), 2.33 (d, J = 6.0 Hz, 2H), 3.91 -3.93 (m, 2H), 6.44 (s, 1H), 6.93 (t,

121 J= 7.2 Hz, 1H), 7.05 (q, J= 8.0 Hz, 2H),

7.15-7.21 (m, 2H), 7.33 (d, J= 8.0 Hz, 2H), 7.40-7.42 (m, 2H), 7.47 (d, J= 8.0 Hz, 1H), 7.74 (s, 1H), 11.07 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.80 (t, J = 6.0 Hz, 2H), 2.09 (s, 6H), 2.31 (t, J= 7 ^' .20 Hz, 2H), 3.95 (t, J = 6.0 Hz, 2H), 6.96 (t, J =

122 8.0 Hz, 1H), 7.06 (d, J= 8.0 Hz, 1H), 7.1 1- 7.17 (m, 2H), 7.21 (t, J= 8.0 Hz, 1H), 7.48 (s, 1H), 7.66 (s, 2H), 7.71 (d, J= 8.0 Hz, 1H), 8.22 (d, J= 6.0 Hz, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.81 (t, J = 6.0 Hz, 2H), 2.10 (s, 6H), 2.33 (t, J= 7.2 Hz, 2H), 3.94 (t, J= 7.2 Hz, 2H), 6.95 (s, 2H), 7.05 (d, J= 8.0 Hz, 1H), 7.12 (d, J = 8.0 Hz,

123 OCO 1H), 7.18-7.23 (m, 2H), 7.40 (d, J= 8.0 Hz,

1H), 7.50 (d, J= 8.0 Hz, 1H), 7.67 (s, 1H), 7.70 (d, J= 8.4 Hz, 1H), 8.00 (s, 1H), 11.99 (s, 1H) Cmpd# Structure NMR data

¾ NMR (DMSO-i e, 400 MHz) 51.36-1.49 (m ,3 H), 1.80-2.01 (m, 5H),2.11-2.31 (m,

124 IH), 3.089 (bs, IH), 6.70-6.77 (m,2 H), 7.00- 7.04 (m, IH), 7.20(s, IH), 7.30(s, IH), 7.77 (bs, 3H), 8.18(s, IH), 9.04 (s, IH)

¾ NMR (DMSO-de _, 400 MHz) δ 1.16 (s, IH), 1.24 (d, J= 10.8 Hz, 2H),1.34 (d, J = 5.6 Hz, 2H), 1.83(s, 2H), 1.86 (d, J =8.4 Hz,

125 1H),2.02 (d, J= 12 Hz, IH), 2.65 (s, 2H),

6.87 (t, J= 6.8 Hz, IH), 6.92 (d, J= 8 Hz, IH), 6.98 (d, J= 7.6 Hz, IH), 7.07 (t, J= 7.6 Hz, 1H),7.12 (s,lH); 7.38 (s,lH), 8.04(s,lH)

¾ NMR (DMSO-i 6, 400 MHz) δ 1.27-1.32 ( m, 2H), 1.76-1.82(m, 2H), 2.14 (s, 7H), 2.34 (t, J= 6.8Hz, 2H), 4.02(t, J= 8.0Hz, 2H),

126 N 6.86-6.89(m,lH), 6.99 (d, J= 8.4Hz, 1 H),

7.21 (d, J= 8.4Hz, 2H), 7.43 (d, J= 7.6Hz, IH), 7.67-7.69(m,lH),8.01(d, J= 4.0Hz, IH)

¾ NMR (DMSO-de _, 400 MHz) δ 1.81-1.83 (m, 2H), 2.11 (s, 6H), 2.33-2.35 (m, 2H), 3.95-3.97 (m, 2H), 6.95 (t, J= 7.2 Hz, IH),

129 7.06 (d, J= 8.0 Hz, IH), 7.13-7.23 (m, 4H),

7.35 (d, J= 8.0 Hz, IH), 7.44 (s, IH), 7.50 (d,

J= 8.0 Hz, IH), 7.71 (d, J= 8.0 Hz, IH), 8.14 (s, IH), 13.16 (s, IH) Cmpd# Structure NMR data

¾ NMR (DMSO-i e, 400 MHz) 50.84 (bs , H), 1.22(s, 2H), 1.85-1.89 (m, 2H), 2.18 (s, 6H), 2.31 -2.49 (m, 2H), 4.08 (t, J= 7.2Hz,

135 m 2H), 6.81(s, IH), 6.86-6.89 (m,lH), 6.96(t,

J= 7.6Hz, IH), 7.04-7.12 (m, 2 H), 7.35(d, J= 8.4Hz, IH), 7.47 (t, J= 8.0Hz, 2H), 7.58(s, IH).

^X H NMR (DMSO-de _, 400 MHz) δ 1.83 (t, J = 6.0 Hz, 2H), 2.06 (s, 6H), 2.33 (t, J= 7.2 Hz, 2H), 2.72 (t, J= 7.2 Hz, 2H), 3.95 (t, J= 6.8 NH ₂ Hz, 2H), 4.12 (t, J= 7.2 Hz, 2H), 6.44 (s,

136 (ΧΟ IH), 6.96 (t, J= 7.6 Hz, IH), 7.01 -7.07 (m,

2H), 7.12-7.20 (m, 3H), 7.22 (t, J= 8.0 Hz, IH), 7.33 (s, IH), 7.38 (d, J= 8.0 Hz, IH), 7.51 (d, J= 7.2 Hz, 2H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.78-1.89 (m, 2H), 2.10 (s, 6H), 2.33 (t, J= 7.2 Hz, 2H), 3.96 (t, J= 7.2 Hz, 2H), 6.97 (t, J= 7.2

137 Hz, IH), 7.07 (d, J= 8.0 Hz, IH), 7.12-7.24

(m, 5H), 7.47 (d, J = 6.8 Hz, 2H), 7.60 (d, J =

7.2 Hz, IH), 7.90 (s, IH), 7.60-7.99 (m, IH), 12.72 (s, IH)

¾ NMR (DMSO-de _, 400 MHz) δ 1.78 (d, J=8 Hz, IH), 2.06 (s, 2H), 2.42 (s, 3H), 2.94 (s, 2H), 3.29 (s, 2H), 6.87-6.84 (m, IH), 6.90

139 (d, J=8.6 Hz, IH), 6.97 (d, J=7.2 Hz, IH),

7.06 (t, J= 8 Hz, IH), 7.15 (s,lH), 7.36 (s,lH), 7.48 (s, IH), 8.0 (s, IH), 9.44 (s, IH) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.41-1.48 (m, 4H), 1.90-1.93 (d, J= 12 Hz, 2 H), 2.063 (t, J=12 Hz, 2H), 2.30 (t, J=4 Hz, 2H), 2.58- 2.61 (m, 2H), 2,82-2.84 (d, J=8 Hz, 2H),

140 a: k„ 5.26-5.28 (d, J=8 Hz, 1H), 6.54 (s, 1H), 6.58

(s, 1H), 6.80-6.82 (m, 1H), 6.84-6.88 (m, 1H), 6.90-6.95 (m, 1H), 7.01-7.05 (m, 1H), 8.02 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.25-1.32 (m, 2H), 1.89 (d, J = 11.2 Hz, 2H), 2.01-2.02

^NH (m, 1H), 2.49-2.57 (m, 2H), 2.97 (d, J= 12

141 Hz, 2H), 3.29 - 3.33(m, 1H), 6.53 (s, 2H),

6.58 (s, 2H), 6.82 (t, J= 7.2 Hz, 1H), 6.90 (d, J= 8 Hz, 1H), 6.95 (d, J= 7.6 Hz, 1H), 7.03 (t, J= 7.2 Hz, 1H), 8.02 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.78-1.85 (m, 2H), 2.08 (s, 6H), 2.32 (t, J= 7.2 Hz, 2H), 3.97 (t, J = 6.8 Hz, 2H), 6.98 (t, J= 7.6 Hz, 1H), 7.08 (d, J= 8.0 Hz, 1H), 7.16-7.18

143 (m, 2H), 7.23 (t, J= 7.6 Hz, 1H), 7.42 (t, J =

7.6 Hz, 1H), 7.51 (d, J= 7.2 Hz, 1H), 7.79 (d, J= 2.4 Hz, 1H), 7.87-7.89 (m, 1H), 7.99 (d, J = 8.0 Hz, 1H), 8.09 (d, J= 8.0 Hz, 2H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 2.05-2.07 (m, 2H), 2.75 (s, 6H), 3.15-3.17 (m, 2H), 4.00 (t, J= 6.8 Hz, 2H), 6.98 (s, 1H), 7.02 (d, ° J= 8.0 Hz, 1H), 7.10 (d, J= 8.4 Hz, 1H),

151 7.17 (d, J = 8.4 Hz, 1H), 7.22-7.27 (m, 2H),

7.41 (d, J= 8.1 Hz, 1H), 7.51 (d, J= 8.4 Hz, 1H), 7.69-7.75 (m, 2H),8.01 (s, 1H), 9.29 (s, 1H), 12.02 (s, 1H).

¾ NMR (DMSO-de _, 400 MHz) δ 1.82-1.84 (m, 2H),1.97-1.99 (m, 1H), 2.06 (s, 6H), 2.48 (t, J= 7.2 Hz, 2H), 3.93 (t, J= 7.2 Hz, 2H),

152 6.55-6.60 (m, 2H), 6.78 (t, J= 2.0 Hz, 2H),

6.94 (t, J= 7.2 Hz, 1H), 7.03-7.22 (m, 5H),

7.57 (s, 1H), 7.61 (d, J= 7.6 Hz, 1H), 11.06 (s, 1H)

¾ NMR (DMSO-de, 400 MHz) δ 1.22 (s, 6H), 1.89 (s, 2H), 3.87 (s, 3H), 3.94 (t, J = 6.0, 2H), 6.86 (d, J= 8.8, 1H), 6.96 (t, J =

153 C C , 6.2, 2H), 7.108-7.042 (m,2H), 7.215-7.162

(m,2H), 7.449 (s, 1H), 7.48 (d, J= 8.4 Hz, 1H), 7.96 (d ,J= 6 Hz, 1H), 8.43(d ,J= 2.8 Hz, 1H).

¾ NMR (DMSO-de _, 400 MHz) δ 0.1.82- 1.85 (m, 1H), 2.10-2.15 (m, 2H), 2.99-3.22 (m,

155 7H), 6.84-6.89 (m, 2H), 6.98 (d, J= 7.6 Hz,

1H), 7.05 (t, J= 7.6 Hz, 1H), 7.17 (s, 1H), 7.34 (s, 1H), 7.91-8.04 (m, 4H). Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 0.94-0.97 (m, 1H), 1.18-1.26 (m, 3H), 1.74-1.83 (m,

156 4H), 1.96 (d, J = 11.2 Hz, 1H), 2.49-2.65 (m,

4H), 6.65-6.67 (m, 3H), 6.73 (s, 1H), 6.77- 6.81 (m, 1H), 7.26 (s, 1H), 7.87 (s, 1H).

¾ NMR (DMSO-de _, 400 MHz) δ 2.49 - 2.62(m, 2H), 2.81-2.92 (m, 2H), 3.43 (s, 2H), 3.59 (d, J= 8.8 Hz, 1 H), 3.69-3.74 (m, 3H),

157 4.03 (d, J= 10 Hz, 2H), 6.90 (d, J= 6.4 Hz,

2H), 7.01 (d, J= 7.2 Hz, 1H) , 7.09 (t, J = 7.2 Hz, 1H) , 7.20 (s, 1H), 7.40 (S, 1H), 7.56 (br s, 2 H), 7.97 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.22-1.32 (m, 4H), 1.45-1.55 (m, 2H), 1.55 (bs, 1H), 1.78 (m, 2H), 1.88-1.91 (m, 2H), 2.95 (s,2H),

158 5.50 (s, 1H), 6.50 (s, 1H), 6.55 (s, 1H), 6.81- 6.84 (t, J=7.2 Ηζ,ΙΗ), 6.89-6.91 (d, J=7.6 Ηζ,ΙΗ), 6.94-6.96 (d, J=7.6 Hz, 2H), 7.02- 7.05 (t, J= 7.2 Hz, 1H), 8.03 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 2.73-2.81 (m, 2H), 2.73-2.81 (m, 2H), 2.92-2.94 (m, 1H), 2.97-2.02 (m, 2H), 3.35 (t, J= 10.4 Hz, 1H), 3.67 (d, J= 7.2 Hz, 1H), 3.83 (d, J =

159 10.4 Hz, 1H), 5.39 (br s, 1H), 6.58 (s,

2H), 6.82 (t, J= 7.2 Hz, 1H), 6.89 (d, J = 7.6 Hz, 1H), 6.95 (d, J= 7.2 Hz, 1H), 7.04 (t, J= 6.8 Hz, 1H), 7.97 (s, 1H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 0.99(d, J=12.4Hz, 2H), 1.22 (s, 2H), 1.4 (d,

J=13.2Hz, 2H), 1.543(s, 1H), 1.8(d, J=12.4

160 Hz, 2H), 1.96(d, J=9.6 Hz, 2H), 2.65 (s, 1H), 6.66 (d, J=7.2Hz, 2H), 6.744(s, 1H), 6.67 (d,

J= 5.6 Hz, 1H), 7.24 (s, 1H), 7.65 (bs, 2H), 7.82 (s, 1H), 9.11 (s, 1H)

^X H NMR (DMSO-de _, 400 MHz) δ 0.95-0.98 (m, 1H), 1.62-1.32 (m, 4H), 1.75-1.99 (m,

161 3H), 2.60 (s, 1H), 3.46 (t, J= 4.0 Hz, 1H),

3.68 (d, J= 7.2 Hz, 1H), 6.80-6.90 (m, 3H), 6.96 (d, J= 7.6 Hz, 1H), 7.04 (t, J= 8.0 Hz, 1H), 7.14 (s, 1H), 7.71 (s, 1H), 9.14 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.12 (s, 2H), 1.30 (s, 2H), 1.46 (s, 1H), 1.62 (s, 3H),

162 1.77 (s, 2H), 1.88 (s, 2H), 2.16 (d, J=7.6 Hz,

S ^L _CFj

2H), 2.66 (s, 1H), 2.79 (s, 2H), 6.83 (s, 2H), 7.01-6.93 (m, 4H), 7.09 (s, 1H), 7.41 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ, 0.92-1.10 (

NH, m ,2H), 1.44-1.59 (m ,2H), 1.71 -1.98 (m,3H), ° ^

163 2.11-2.14 (m ,2H), 2.49-2.95 (m,8 H), 6.84- 7.10 (m, 5H),7.41 (s,lH),8.17 (bs,lH),9.15 (bs,l H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 0.73-0.76 (m, 2H), 1.27-1.34 (m, 3H), 1.60-1.69 (m, 1H), 1.75-1.78 (m, 3H), 1.93-1.96 (m, 1H),

164 2.66 (b, 2H), 2.91-2.92 (m, 2H), 4.13 (s, 1H),

5.01 (s, 1H), 6.25 (s, 1H), 6.31 (s, 1H), 6.56- 6.62 (m, 2H), 6.74-6.76 (m, 1H), 7.67 (m, 1H)

^X H NMR (400 MHz, DMSO-d6): δ 1.34 - 1.16 (m, 2 H), 1.60 - 1.54 (m, 2 H), 1.79 -

°Y I.69 (m, 2 H), 1.88 (s, 2 H), 2.03 (d, J =

I I .2 Hz, 1 H), 2.49 - 2.25 (m, 3 H), 3.02 (s,

165 1H), 6.88 (q, J= 8.4 Hz, 2 H), 6.98 (d, J =

CX _s i _CF3 7.2 Hz, 2 H), 7.05 (t, J = 7.2 Hz, 1 H), 7.15

(s, 1H), 7.36 (s, 1H), 7.70 (bs, 3 H), 8.03 (s, 1H), 9.39 (s, 1H)

¾ NMR (400 MHz, DMSO-d6): δ 1.80 - 1.13 (m, 11 H), 2.30 (d, J = 10 Hz, 2 H),

167 2.99 (s, 1H), 6.97 - 6.83 (m, 3 H), 7.04 (t, J

= 7.6 Hz, 1 H), 7.11 (s, 1H), 7.39 (s, 1H), 8.3 (bs, 1H), 9.4 (bs, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.27-1.36 (m, 2H), 1.88 (d, J = 11.6 Hz, 2H), 2.56-2.66 ^NH

(m, 2H), 2.98 (d, J = 12 Hz, 2H), 3.28 (b,

169

C A _CF3 1H), 4.13 (s, 1H), 4.84 (d, J= 6.8 Hz, 1H),

6.24 (s, 1H), 6.39 (s, 1H), 6.55-6.64 (m, 3H), 6.74-6.78 (m, 1H), 7.66 (s, 1H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ: 0.80-0.83 (m, IH), 0.95 (d, J = 11.6 Hz, IH), 1.05-1.14 (m, 2H), 1.22 (s, IH), 1.94-1.97 (m, 4H),

170 2.65 (s, IH), 6.65 (bs, IH), 6.83-6.87 (m,

IH), 6.91 (s, IH), 6.97 (d, J= 6.8 Hz, IH), 7.03-7.05 (m, IH), 7.12 (s, IH), 7.37 (s, IH), 8.03 (bs, IH)

^X H NMR (DMSO-de _, 400 MHz) δ: 0.68-0.77 (m, IH), 0.81 -0.87 (m, IH), 0.90-0.96 (m, IH), 1.25 (bs, 2H), 1.63-1.79 (m, 6H), 1.94

171 (d, J= 12.4 Hz, IH), 2.95 (d, J= 6.0 Hz, 2H),

3.16 (d, J= 4.8 Hz, IH), 5.49 (s, IH), 6.50 (s, IH), 6.54 (s, IH), 6.80-6.84 (m, IH), 6.88- 6.90 (m, IH), 7.02-7.05 (m, IH), 8.01 (s, IH)

¾ NMR (DMSO-de _, 400 MHz) δ 1.65 (t,

H j=8.4 Hz, 2H), 1.96 (s, 3H), 2.77 (t, IH), 2.88

(s, IH), 3.19 (

172 JO s, IH), 3.73 (s, IH), 5.22 (s,

IH), 6.65 (d, j=13.2 Hz, 2H), 6.85-6.81 (m, IH), 6.95 (s, J=7.2 Hz, IH), 7.02 (d, J=6 Hz, IH), 8.22 (s, IH)

¾ NMR (DMSO-de _, 400 MHz) δ 1.08-1.22 (m, 2H), 1.74 (d, J = 12 Hz, 2H), 2.95-3.15 (m, 4H), 3.46-3.50 (m, 2H), 3.67-3.70 (m,

173 2H), 5.47 (br s, IH), 6.55 (d, J= 12 Hz,

2H), 6.82 (t, J= 7.6 Hz, IH), 6.90 (d, J= 8 Hz, IH), 6.96 (d, J= 6.8 Hz, IH), 7.03 (t, J = 7.02 Hz, IH), 8.020 (s, IH) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.59-1.63 (m, 1H), 2.00-2.06 (m, 1H), 2.67 (t, J = 8 Hz, 1H), 2.75-2.78 (m, 1 H), 2.88-2.90 (m, 1H), 2.93-3.05 (m, 1H), 3.84 (br s, 1H), 5.46 (d, J

174 = 5.6 Hz, 2H), 6.57 (d, J= 11.2 Hz, 1H) ,

6.83 (t, J= 7.6 Hz, 1H) , 6.89 (d, J= 7.6 Hz, 1H) , 6.95 (d, J= 7.2 Hz, 1H) , 7.01-7.05(m, 1H),7.20 (s, 1H), 8.01 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.88 (d, J = 8.8 Hz, 2H), 2.38 (d, J= 7.2 Hz, 2H), 2.82 (t, J= 8 Ηζ,ΙΗ), 3.26(s,lH), 6.83- 6.90 (m, 1H),

175 6.97 (d, J= 7.6 Hz ,1H), 7.06 (t, J= 7.2 Hz

,1H), 7.11 (s, 1H), 7.39 (s, 1H), 8.07(s,lH), 9.40 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 2.77-2.88 (m, 2H), 3.65-3.72 (m, 2H), 3.77-3.81 (m, 1H), 5.29-5.31 (m, 1H), 6.64 (s, 1H), 6.69 (s,

178 1H), 6.81 -6.85 (t, J= 7.6 Hz, 1H), 6.90 (d, J

= 8 Hz, 1H), ), 6.96 (d, J= 7.6 Hz, 1H), 7.02- 7.06 (t, J= 7.6 Hz, 1H), 8.26 (bs, 1H)

Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.23-1.25 (m, 3H), 1.59 (br s, 1H), 1.79 (d, J= 11.2 Hz, 2H), 1.91 (t, J= 10.8 Hz, 2H), 2.34 (br s, 2H), 2.71 (br s, 2H), 2.87 (d, J= 10.8 Hz, «ϊΎ>

179 2H), 2.98 (br s, 2H), 4.48 (br s, 1H), 5.48 (br s,lH), 6.56 (d, J = 13.2 Hz, 2H), 6.83 (t, J =

7.2 Hz, 1H), 6.91 (d, J= 7.6 Hz, 1H), 6.96 (d, J= 7.6 Hz, 1H), 7.04 (t, J= 7.2 Hz, 1H),

8.03 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.63(d, J=11.2Hz, 6H), 1.72(d, J=8.4Hz, 2H), 1.871 (s, 2H), 1.543(s, 1H), 2.99(bs, 1H), 3.423

180 (bs, 1H), 5.2(d, J=6.4Hz, 1H), 6.54 (d,

J=5.2Hz, 2H), 6.840 (t, J =7.6 Hz, 1H), 6.900-6.956 (m, 2H), 7.053 (t, J=7.2Hz, 1H), 8.213 (s, 1H), 9.117 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.81 (t, J = 6.8 Hz, 2H), 2.11 (s, 6H), 2.34 (t, J= 6.8 Hz, 2H), 3.94 (t, J = 6.8 Hz, 2H), 4.04 (s, 2H),

181 6.85 (s, 1H), 6.95 (t, J= 7.6 Hz, 1H), 7.05 (d,

J= 8.0 Hz, 1H), 7.09-7.23 (m, 4H), 7.38 (d, J = 8.0 Hz, 1H), 7.57 (s, 1H), 7.65-7.95 (m, 4H), 11.51 (s, 1H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 0.75-0.97 (m, 1H), 1.01 (d, J = 10.8 Hz, 1H), 1.25 (br s, 2H), 1.63-1.79 (m, 4H), 1.96 (d, J= 11.6 Hz, 1 H), 2.64(br s, 1H), 2.96 (d, J= 5.6 Hz, 2H),

182 5.50 (s, 1H), 6.53 (d, J= 14 Hz, 2H), 6.83 (t,

J= 7.2 Hz, 1H), 6.88 (d, J= 7.6 Hz, 1H), 6.94 (d, J= 6.8 Hz, 1H), 7.02 (t, J= 7.2 Hz, 1H), 7.98 (s,lH), 8.06 (s,lH)

¾ NMR (DMSO-de _, 400 MHz) δ 1.13-1.31 (m, 3H), 1.87 (d, J = 12.0 Hz, 2H), 2.53-2.64

^NH (m, 2H), 2.96 (d, J = 12.0 Hz, 2H), 5.19 (d, J

184 = 7.6 Hz, 1H), 6.27 (s, 1H), 6.37 (s, 1H), 6.77

S CI (t, J= 7.6 Hz, 1H), 6.84 (t, J= 8.0 Hz, 1H),

6.91 (t, J= 7.2 Hz, 1H), 6.99 (t, J= 7.6 Hz, 1H),7.77 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ, 1.81 ( t, J=6.8 Hz ,2H), 2.23 (s ,3H),2.54-2.64(m,2H), 3.93 (t, J=7.2 Hz ,2H), 6.77 (bs,lH),6.93 (q, J

186 = 7.2 Hz ,2H),7.03(t, J= 6.0 Hz,2H)7.08(d, J

= 8.4 Hz,l H),7.14-7.20 (m,2H),7.33 (d, J = 8.0 Ηζ,ΙΗ), 7.46 (d, J= 7.6 Hz,lH),7.60-7.65 (m,2H)

Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ: 0.84-087 (m, 1H), 0.93-0.96 (m, 1H), 1.02-1.28 (m, 2H), 1.68 (bs, 1H), 1.76-1.83 (m, 2H), 1.88- 1.90 (m, 1H), 3.02 (bs, 4H), 5.64 (s,lH),

187 6.52-6.54 (d, J= 6.0 Hz, 2H), 6.80 (t, J =

7.6Hz, 1H), 6.88 (d, J= 7.6 Hz, 1H), 6.94 (d, J= 6.8 Hz, 1H), 7.00-7.04 (t, J= 6.8 Hz, 1H), 7.73 (bs, 3H), 7.99 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 3.54 (t, J = 7.2 Hz, 2H), 3.61-3.66 (m, 1H), 3.78 (t, J =

0 7.6 Hz, 2H), 3.91-3.99 (m, 1H), 6.82 (d, J =

188 7.6 Hz, 1H), 6.86 (d, J= 6.0 Hz, 1H), 6.95 (d,

J= 7.2 Hz, 1H), 7.03 (d, J= 8.0 Hz, 1H), 7.13 (s, 3H), 7.34 (s, 1H), 8.00 (s, 1H), 9.30 (s, 1H)

¾ NMR (DMSO-de, 400 MHz) δ 0.78-.084 (m, 3H), 1.26-1.33 (m, 4H), 1.40 (bs, 1H), 1.75 (d, J = 11.2 Hz, 2H), 1.83 -1.92 (m, 3H),

189 3.27 (s, 2H), 4.11 (s, 1H), 5.51 (s, 1H), 6.48

(s, 1H), 6.53 (s, 1H), 6.81 (t, J= 7.2 Hz, 1H), 6.89 (d, J= 7.6 Hz, 1H), 6.94 (d, J= 7.2 Hz, 1H), 7.02 (t, J= 7.2 Hz, 1H),8.01 (s, 1H)

Cmpd# Structure NMR data

¾ NMR (DMSO-de, 400 MHz) δ 1.10 (d, J=6.4 Hz, 3H), 1.48 (s, 2H), 1.72 (d, J=14.0 Hz, 4H), 1.86 (s, IH), 1.96 (s, IH), 3.4 (s,

190 2H), 5.21 (d, J=8 Hz, IH), 6.52 (d, J=10 Hz,

2H), 6.81 (d, J=7.6 Hz, IH), 6.89 (d, J=7.2 Hz, IH), 6.94 (d, J=7.6 Hz, IH), 7.03 (t, J=7.6 Hz, IH), 8.04 (s, IH)

^X H NMR (DMSO-de, 400 MHz) δ 1.07 (d, J=12.0 Hz, IH), 1.52-1.45 (m, 3H), 1.78 (s, IH), 1.87 (s, J=12.8 Hz, IH), 2.06 (s, IH),

191 2.20 (d, J=12.4 Hz, IH), 2.85 (s, IH), 3.46 (s,

IH), 7.18 (t, J=7.6 Hz, IH), 7.23 (s, IH), 7.31

(t, J=7.6 Hz, 2H), 7.54 (s, IH), 7.63 (d, J=8.4 Hz, IH)

¾ NMR (DMSO-de, 400 MHz) δ 0.99-1.08 (m, 2H), 1.15-1.21 (m, 3H), 1.27(br s, IH), 1.76 (d, J= 12 Hz, 2 H), 2.01 (d, J= 11.2 Hz,

192 2 H), 3.21-3.27 (m, 2H), 6.51 (s, 1 H), 6.55

(s, 1 H), 6.82 (t, J= 6.8 Hz, IH), 6.88 (d, J = 8 Hz, IH), 6.94 (d, J= 7.2 Hz, IH), 7.03 (t, J= 7.2 Hz, IH), 7.97 (s,lH)

¾ NMR (DMSO-de, 400 MHz) δ 3.53-3.57 (m, 2H), 3.91 -3.97 (m, 2H), 4.26-4.27 (m,

Γ^ΝΗ

IH), 5.93-5.94 (m, IH), 6.30 (s, IH), 6.61 (s,

193 ooX IH), 6.72-6.86 (m, 2H), 6.93 (d, J= 7.6 Hz,

IH), 7.01 -7.05 (t, J= 7.6 Hz, IH), 7.96 (s, IH) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.21-1.25 (m, IH), 1.48 (d, J = 8.4 Hz, 2 H), 1.66 (br s, IH), 1.87 (br s, IH), 1.98 (br s, IH), 2.18 (d, J= 10 Hz, IH), 2.33 (br s, IH), 2.64-2.67 (m,

203 3H), 2.94 (d, J= 10 Hz, IH), 3.07 (br s,

IH), 6.82-6.88 (m, 2H), 6.96 (d, J= 7.6 Hz, I H), ), 7.04 (t, J= 7.2 Hz, 1 H), 7.11 (s, 1 H), 7.34 (s, 1 H)

¾ NMR (DMSO-de _, 400 MHz) δ, 0.92-1.07 ( m, 2H),1.31-1.35 (m, 2H),1.52-1.75 (m,4H), 1.91(d, J=12.4 Hz,lH),2.06 (d, J=11.6 Ηζ,ΙΗ), 3.13-3.72 (m, 0.5H, m, 0.5 H), 5.10

204 (d, J=6.0Hz,0.5 H), 5.23 (d, J=6.0Hz,0.5 H),

6.52 (s,lH), 6.56 (d, J= 11.6 Hz,l H), 6.80 (t, J= 7.2 Ηζ,ΙΗ), 6.86 (d, J = 8.0 Ηζ,ΙΗ), 6.93 (d, J= 7.6 Hz,lH),7.04 (t, J= 7.6 Hz, IH), 7.97 (s,0.5 H), 8.05 (s,lH)

¾ NMR (DMSO-de _, 400 MHz) δ 5.43 (s, 2H), 6.52 (s, IH), 6.72 (d, J=5.2 Hz, 2H), 6.88 (d, J=8.4 Hz, IH), 6.94 (t, J=7.6 Hz,

205 IH), 7.02 (t, J=7.6 Hz, IH), 7.31 (d, J=8.0

Hz, IH), 7.43 (d, J=4 H, IH), 7.45 (s, IH), 7.49 (d, J=8.4 Hz, IH), 7.92 (s, IH), 11.3 (s, IH) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.20-1.25 (m, IH), 1.33-1.37 (m, 2H), 1.77-1.80 (m, 2H), 1.91 (s, IH), 2.03-2.06 (d, J=12 Hz,

211

IH), 2.64-2.69 (m, IH), 2.89-291 (m, IH),

8.05-8.12 (m, 2H), 8.28 (s, IH), 8.31-8.34 (dd, IH), 8.40-8.42 (dd, IH), 8.91 (s, IH)

¾ NMR (400 MHz, DMSO-d6):5 1.58 (q, J = 10 Hz, 2 H), 2.00 - 1.96 (m, 1 H), 2.08 (d, J= 13.6 Hz, 2 H), 3.05 (d, J= 10.8 Hz, 2 H), 3.37 (bs, 2H), 3.66 (s, IH), 5.33 (s, IH),

212 6.64 (s, IH), 5 6.71 (d, J= 17.6 Hz, 2 H),

6.96 - 6.90 (m, 2 H), 7.03 (t, J= 7.2 Hz, 1 H), 7.32 (d, J= 8 Hz, 1 H), 7.45 (s, 2 H), 7.52 (d, J= 8.4 Hz, 1 H), 8.10 (s, 1 H), 8.29 (bs, 1 H), 8.47 (bs, 1 H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.31-1.33 (m, 2H), 2.89 (t, J= 6.4 Hz, 2H), 3.92 (t, J = 6.4 Hz, 2H), 6.78 (s, IH), 6.94 (t, J= 7.6 Hz,

213 2H),7.02-7.21 (m, 5H), 7.33 (d, J= 7.6 Hz, IH), 7.46 (d, J= 7.2 Hz, IH), 7.62-7.68 (m,

2H), 11.3 (s, IH)

Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.37-1.45 (m, 2H), 1.77-1.90 (m, 2H), 2.03-2.06 (m, 2H), 2.26-2.29 (m, 2H), 2.58-2.64 (m, 2H), 2.82 (d, J= 12.0 Hz, 2H), 3.20-3.27 (m, 1H),

214 5.23 (d, J= 6.4 Hz, 1H), 6.22 (s, 1H), 6.38 (s,

1H), 6.76 (t, J= 8.4 Hz, 1H), 6.85 (d, J= 8.0 Hz, 1H), 6.91 (d, J= 8.0 Hz, 1H), 6.99 (t, J = 8.0 Hz, 1H), 7.82 (s, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.31 (m, 2H), 1.39-1.44 (m, 2H), 1.65 (s, 1H), 1.77- 1.91 (m, 2H), 2.04-2.20 (m, 2H), 2.30 (s,

215 2H), 2.57-2.64 (m, 2H), 2.82 (d, J= 9.6 Hz,

2H), 5.31 (d, J= 5.6 Hz, 1H), 6.67 (s, 1H), 6.74 9s, lH),6.79-6.83 (m, 1H), 6.87-6.91 (m, 2H), 6.92-7.03 (m, 1H),8.19 (s, 1H)

¾ NMR (DMSO-de, 400 MHz) δ, 1.52( d, J=11.6 Hz ,2H), 1.88-1.92 (m,2H),2.10(t, J=11.2 Hz ,2H), 2.37- 2.64(m,10H),2.80(bs,3H) 2.89 (d, , J= 12.0

216 Hz ,3H),5.49 (d, J= 6.4 Hz

,lH),6.53(s„lH),6.58(s,l H),6.80- 6.82(m,lH),6.92(d, J= 7.6 Hz,lH), 7.01(d, J = 3.6 Hz,lH), 8.19(s, 1H) Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.42-1.45 (m, 2H), 1.88 (bs, 2H), 2.07 (d, J=l 1.2 Hz, 2H), 2.30-2.33 (m, 2H), 2.65 (d, J= 8 Hz,

217 2H), 2.81 (bs, 2H), 5.32 (d, J=6.4 Hz, 1H),

6.54 (s, 1H), 6.59 (s, 1H), 6.93 (d, J= 8.4 Hz, 1H), 7.39-7.43 (m, 2H), 8.51 (bs, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.5 (d, J=10.8Hz, 2H), 1.85 (d, J=9.2Hz, 2H), 2.107(t, J=10Hz, 2H), 2.2(t, J=5.6Hz, 2H), 2.5(d, J=6 Hz, 2H), 2.77 (d, J=10 Hz, 2H),

218 2.970(s, 1H), 3.122(s, 3H), 3.272(s, 2H),

CC _{S F3} 5.144(d, J=8.4Hz, 1H), 6.715 (s, 1H),

6.744(s, 1H), 7.044 (t, J=7.6Hz, 1H), 7.171 (d, J=7.6Hz, 1H), 7.23 (t, J=7.2Hz, 1H), 7.3015 (d, J=7.6Hz, 1H)

¾ NMR (DMSO-de _, 400 MHz) δ, 0.95 ( d, J=6.4 Hz ,2H), 1.42 (d, J=10.0 Hz ,2H),1.90 (d, J=11.6 Hz ,2H), 2.37-2.70 (m,10H),2.83

219 (d, J=11.6 Hz, 2H),5.23 (d, J= 6.4 Hz ,1H),

6.53 (s,lH), 6.57 (s,l H),6.81 (t, J= 7.6 Ηζ,ΙΗ), 6.86 (d, J= 7.6 Ηζ,ΙΗ), 6.93 (d, J = 7.6 Ηζ,ΙΗ), 7.02 (t, J= 7.6 Hz), 7.97 (s, 1H)

Cmpd# Structure NMR data

¾ NMR (DMSO-de _, 400 MHz) δ 1.01 (d, J = 11.6 Hz, 1 H), 1.40-1.45 (m, 2H), 1.77 (d, J = 10.8 Hz, 2H), 1.93-1.98 (m, 2H), 2.11 -2.23 (m, 2H), 2.60-2.64 (m, 1H), 2.68 (br s, 1H),

220 2.71 (br s, 1H), 2.86 (br s, 2H), 5.47 (br s,

1H), 6.50 (s,lH) , 6.54 (s,lH) ,6.81 (t, J = 7.2 Hz, 1 H), 6.88 (d, J = 7.2 Hz, 1 H), 6.94 (d, J = 6.8 Hz, 1 H), 7.02 (t, J = 7.2 Hz, 1 H), 8.02 (s, 1 H)

¾ NMR (DMSO-de _, 400 MHz) δ 1.21 (br s, NH 2H), 1.63-1.69 (m, 2H), 1.74-1.84 (m, 2H),

HN ' 1.98-2.01 (m, 2H), 3.06 (t, J = 11.2 Hz, 2H),

221 σΐιΧ, 3.08-3.18 (m, 2H), 3.66 (br s, 1H), 6.55 (d, J

= 12.4 Hz, 2H), 6.80 (t, J = 7.2 Hz, 1H), 6.92-7.03 (m, 3H), 8.20 (s, 1H)

¾ NMR (DMSO-de, 400 MHz) δ 1.99 (t, J = 7.2 Hz, 2H), 2.91 (m, 4H), 4.02 (t, J = 7.2 Hz, 2H), 4.35 (t, J = 7.6 Hz, 2H), 6.47 (s, 1H),

222 7.00 (t, J = 7.2 Hz, 1H), 7.08 (t, J = 8.0 Hz,

2H), 7.16-7.27 (m, 4H), 7.36 (d, J = 9.6 Hz, 2H), 7.55 (d, J = 8.0 Hz, 2H), 7.70-7.76 (m, 6H).

NMR (DMSO-de _, 400 MHz) δ 1.26-1.31 (m, 2H), 1.85-1.88 (m, 2H), 2.48-2.58 (m, 2H), 2.93-2.96 (m, 2H), 2.31-2.39 (m, 1H), 5.22

223 (d, J=6.8 Hz, 1H), 6.53 (s, 1H), 6.58 (s, 1H),

6.85 (d, J = 8 Hz, 1H), 7.04-7.06 (m, 2H), 8.11 (bs, 1H).

Cmpd# Structure NMR data

IH NMR (400 MHz, DMSO-d6) δ 1.01 -1.07 (m, IH), 1.22-1.34 (m, 4H), 1.77-1.83 (m, 2H), 1.89 (d, J_= 10.3Hz, IH), 2.05 (d, J_=

257 11.7Hz, IH), 2.52-2.59 (m, IH), 2.69-2.75

(m, IH), 6.91 (d, J_= 5.3Hz, IH), 7.19 (s, IH), 7.43 (s, IH), 7.98 (s, IH), 8.07 (d, J_= 5.3Hz, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.22-1.28 (m, 2H), 1.32 (t, J_ = 5Hz, 3H), 1.81 (t, J_ = 6.7Hz, 2H), 2.71 (t, J_= 6.6Hz, 2H), 4.02 (t, J =6.9Hz, 2H), 4.27 (dd, J=7Hz, J=7Hz, 2H),

258 6.84 (s, IH), 6.97 (t, J=7.3Hz, IH), 7.07 (t, J

-^^J,

= 7.7Hz, IH), 7.18 (t, J=9.5Hz, 2H), 7.36 (d, J=8Hz, IH), 7.49 (d, J=7.6Hz, IH), 7.65- 7.71 (m, 3H), 7.80 (dd, J_ = 2Hz, J_= 8.6Hz, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.23 (s, IH), 1.41 (s, 2H), 1.60-1.63 (m, 6H), 1.80 (s, IH), 1.92 (s, IH), 2.05 (d, J_= 11.9Hz, 2H),

259 2.96 (t, J_= 11.9Hz, 2H), 3.27 (d, J_= 3.6Hz,

C _{S CF3} 3H), 3.53 (s, IH), 3.81 (s, 2H), 4.92 (d, J_=

6.8Hz, IH), 5.41-5.44 (m, IH), 6.48 (s, IH), 6.56 (s, IH)

Cmpd# Structure NMR data

IH NMR (400 MHz, DMSO-d6) δ 1.98-2.01 (m, 2H), 2.90-2.95 (m, 2H), 4.07 (t, J_=

H ₂ N 6.6Hz, 2H), 6.86 (s, IH), 6.97 (t, J_= 7.6Hz, TFA 1

IH), 7.07 (t, J_= 7.6Hz, IH), 7.15 (dd, J_=

262

0 -J ₎ 8.4Hz, J_= 11.3Hz, 2H), 7.36 (d, J=8Hz,

IH), 7.49 (d, J=7.8Hz, IH), Ί .69-1.13 (m, 5H), 7.78 (dd, J_= 1.6Hz, J=8.4Hz, IH), 11.46 (s, IH), 12.49 (s, IH)

IH NMR (400 MHz, DMSO-d6) 51.14-1.41 (m, 4H), 1.81-1.89 (m, 3H), 2.03 (d, J

H =10.5Hz, IH), 2.55 (s, IH), 2.95 (s, IH), 6.85

263 r ^NH2

(dd, J=4.8Hz, J=7.4Hz, IH), 7.19 (s, IH), 7.40 (d, J=7.5Hz, IH), 7.53 (s, IH), 7.88 (d, J=4.6Hz, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.22 (s, IH), 1.33-1.41 (m, 2H), 1.89-1.91 (m, 2H), 2.658-2.70 (m, IH), 3.05 (d, J=12.7Hz, 2H),

264 3.44 (d, J=5.4Hz, IH), 6.65 (s, IH), 6.61 (s,

IH), 6.82 (dd, J=5Hz, J=7.5Hz, IH), 7.34 (d, J=7.4Hz, IH), 7.86 (d, J_ = 4.9Hz, IH), 8.36 (s, IH), 8.74 (s, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.93-1.95

H ₂ N (m, 2H), 2.85-2.88 (m, 2H), 3.18 (s, 4H), 2HCI J

3.26 (s, 4H), 3.97 (t, J=7.2Hz, 2H), 6.87 (s,

265

2H), 7.00-7.02 (m, IH), 7.15 (d, J_= 8.4Hz, IH), 7.49-7.53 (m, 2H), 7.77 (s, 2H), 8.89 (s, 2H) Cmpd# Structure NMR data

IH NMR (400 MHz, DMSO-d6) δ 1.93 -1.96 (m, 2H), 2.86 (d, J_ = 5.1Hz, 2H), 3. 10-3.19 (m, 4H), 3.36 (d, J = 12.1Hz, 4H), 3.63-3.69

266 C x (m, 2H), 3.82 (s, 2H), 3.96 (t, J_= 6.5Hz, 2H),

6.91 (s, 2H), 7.02 (d, J_= 8.5Hz, IH), 7.16 (d, J_= 8.2Hz, IH), 7.43 -7.59 (m, 2H), 7.87 (s, 2H), 8.30 (s, 2H)

IH NMR (400 MHz, DMSO-d6) δ 1.97-2.00 (m, 2H), 2.95-2.96 (m, 2H), 4.08 (t, J_=

H ₂ N

6.7Hz, 2H), 6.90 (s, IH), 6.98 (t, J_= 7.4Hz, TFA J IH), 7.09 (t, J_= 7.4Hz, IH), 7.16 (d, J_=

267

6. 1Hz, IH), 7.24 (d, J_= 8.4Hz, IH), 7.37 (d, J_ = 8Hz,lH), 7.50 (d, J_= 7.8Hz, IH), 7.71 - 7.74 (m, 4H), 8.31 (s, IH), 8.36 (d, J_ =

5.7Hz, IH), 1 1.49 (s, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.42-1.50 (m, 2H), 1.87 (d, J =10.8Hz, 2H), 2.08 (t, J_= 1 lHz, 2H), 2.33 (t, J_= 6.2Hz, 2H), 2.66 (t, J

268 Γ ^Ν Τ ^Ν Υ Ί = 6.3Hz, 2H), 2.83 (d, J_= 1 1.8Hz, 2H), 5.69

(d, J_= 6.7Hz, IH), 6.57 (d, J =l l Hz, 2H), 6.81 (dd, J=5Hz, J=7.4Hz, I H), 7.33 (d, J =7.4Hz, IH), 7.86 (d, v = 4.8Hz, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.05 (t, J =7Hz, IH), 1. 19 (s, IH), 1.63 (d, J=12.4Hz, 2H), 2.05 (d, J =13Hz, 2H), 3.01 (t, J_=

269 13.9Hz, 2H), 3.13-3.48 (m, 16H), 6.58 (s,

IH), 6.66 (s, 2H), 6.71 (dd, J_= 2.4Hz, J_= 8.7Hz, IH), 6.82-6.89 (m, IH), 8.33-8.39 (m, 5H), 8.76 (s, 2H) Cmpd# Structure NMR data

IH NMR (400 MHz, DMSO-d6) δ 0.83-0.86 (m, IH), 0.89 (d, J=6.4Hz, 6H), 1.22-1.26 (m, IH), 1.74-1.78 (m, IH), 1.94-1.97 (m,

274 2H), 2.11 (d, J_=\ 1.9Hz, 4H), 2.95 (s, 4H), 3.10-3.18 (m, 2H), 3.30-3.39 (m, 7H), 6.63

(d, J=21.3Hz, 2H), 7.02 (s, IH), 7.42 (s, 2H), 8.26 (s, 3H)

IH NMR (400 MHz, DMSO-d6) δ 1.22 (s, 7H), 1.94 (s, 2H), 2.10 (s, 2H), 3.18 (d, J

275 =10.3Hz, 2H), 3.31-3.45 (m, 7H), 3.63 (s, 4H), 3.83 (s, IH), 4.27 (t, J=7.6Hz, 2H),

6.58-7.12 (m, 4H), 8.34 (s, 4H), 10.93 (s, IH)

IH NMR (400 MHz, DMSO-d6) δ 0.82 (d, J =6.7Hz, IH), 1.06-1.09 (m, IH), 1.22 (s, 4H), 1.34-1.37 (m, 2H), 1.58 (d, J=13.5Hz, IH), J 1.75 (d, J=15Hz, 2H), 1.98-2.05 (m, 3H),

2.87 (t, J=6.5Hz, 2H), 3.08-3.13 (m, 2H),

275

A, 'S 3.21 (s, 2H), 3.34 (s, 4H), 3.63 (s, 2H), 3.75

(s, IH), 3.99 (t, J=5.5Hz, 2H), 6.88 (d, J =8Hz, IH), 7.03 (d, J=8.7Hz, IH), 7.13 (d, J =8.8Hz, IH), 7.76 (s, IH), 8.42 (d, J=8.3Hz, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.26 (t, J =7Hz, 3H), 1.89-1.91 (m, 2H), 2.79 (t, J

=7.1Hz, 2H), 3.98 (t, J=6.8Hz, 2H), 4.24

276 (dd, J=7Hz, J=l 4.1Hz, 2H), 7.04 (d, J

=9.1Hz, IH), 7.15 (d, J=8.7Hz, IH), 7.38- 7.40 (m, 2H), 7.14 (d, J=1.9Hz, IH), 7.75 (dd, J=1.9Hz, J=8.5Hz, IH) Cmpd# Structure NMR data

IH NMR (400 MHz, DMSO-d6) δ 1.91 -1.94 (m, 2H), 2.85 (t, J=6.6Hz, 2H), 3.22-3.29 (m, 3H), 3.30-3.36 (m, 7H), 3.93 (t, J

277 =6.3Hz, 2H), 6.86-6.90 (m, 2H), 7.01 (d, J

=8.4Hz, IH), 7.08 (d, J=8.7Hz, IH), 7.61 (s, IH), 7.74 (d, J=8.3Hz, IH), 7.86 (s, 3H), 8.29 (s, 3H)11.24 (brs, IH)

IH NMR (400 MHz, DMSO-d6) δ 8.36 (s, 2H), 7.92 (s, 2H), 7.36-7.34 (m, 2H), 6.97 (t, J

J=8.1Hz, IH), 6.89 (d, J=10Hz, 2H), 3.90

278 (t, J=6.5Hz, 2H), 3.75 (d, J=l 1.5Hz, 2H),

3.65-3.56 (m, 2H), 3.48-3.35 (m, 4H), 3.19- 3.11 (m, 3H), 2.85 (s, 2H), 1.95-1.91 (m, 2H), 1.28-1.22 (m, 2H)

IH NMR (400 MHz, DMSO-d6) δ 1.08 (d, J J

=6.1Hz, 6H), 1.91 (s, 2H), 2.22 (s, 2H), 2.85

279 XXXI (s, 2H), 3.47 (d, J=10.9Hz, 2H), 3.69 (s, 4H),

V 6.86 (s, IH), 6.95 (d, J=6.7Hz, 2H), 7.35(s,

2H), 7.81 (s, 3H)

11 H NMR (400 MHz, DMSO-d6) δ 1.92- 1.95 (m, 2H), 2.19 (d, J=12.7Hz, 2H), 3.12- 1 J 3.15 (m, 2H), 3.32-3.48 (m, 5H), 3.66 (d, J

280 =10.8Hz, 2H), 6.72-6.79 (m, 2H), 6.80 (t, J

=7Hz, IH), 6.92-7.01 (m, 7H), 8.23 (s, 4H), 10.11 (bs, IH), 10.95 (bs, IH) (s,

Cmpd# Structure NMR data

1H NMR (400 MHz, DMSO-d6) δ 1.26 (t, J =7Hz, 3H), 1.88-1.94 (m, 2H), 2.08 (d, J =13.4Hz, 2H), 3.13 (q, J=l l. lHz, 2H), 3.30

285 (s, 5H), 3.60-3.67 (m, 3H), 4.25 (dd, J=7Hz,

J=14Hz, 2H), 6.59-6.64 (m, 2H), 7.68 (d, J =1.7Hz, 1H), 8.10-8.38 (m, 4H), 9.30 (s, 1H), 11.11 (brs, 1H)

1H NMR (400 MHz, DMSO-d6) δ 8.39 (s,

V 2H), 8.00 (s, 2H), 7.73-7.55 (m, 2H), 7.33-

286 Y 7.64 (m, 4H), 3.88 (s, 2H), 3.35-3.12 (m,

13H), 2.85 (s, 4H), 1.94 (s, 2H), 1.77 (s, 1H), 0.89 (s, 6H)

1H NMR (400 MHz, DMSO-d6) δ 1.89-1.96 (m, 2H), 2.16 (d, J=12.2Hz, 2H), 3.14 (d, J =10.5Hz, 2H), 3.31-3.37 (m, 5H), 3.65 (d, J

287 =10.5Hz, 3H), 6.78 (d, J=6.7Hz, 1H), 6.80- 6.98 (m, 3H), 7.00-709 (m, 1H), 7.70 (brs,

1H), 8.14 (s, 1H), 8.27 (s, 3H), 10.93 (brs, 1H)

1H NMR (400 MHz, DMSO-d6) δ 1.79 (s, 4H), 1.92 (d, J=11.4Hz, 2H), 2.12 (d, J

1 J =12.4Hz, 2H), 3.13 (d, J=10.1Hz, 2H), 3.30-

288 3.39 (m, 9H), 3.61 (d, J=10.2Hz, 4H), 6.40- o 6.47 (m, 1H), 6.58 (s, 1H), 6.78 (t, J=6.6Hz,

1H), 6.91 -7.02 (m, 3H), 8.11 (s, 1H), 8.31 (s, 3H), 8.59 (s, 1H), 10.94 (brs, 1H)

8.55(s,lH), 10.82 (brs, IH)

8.25 (brs, 3H), 8.67 (s, IH) 9.22 (brs, 2H) Cmpd# Structure NMR data

IH NMR (400 MHz, DMS0-d6) δ 1.93 (s, IH), 2.11 ( s,lH), 2.77 ( s,lH),3.14-

299 3.17(m,2H), 3.27-3.39 (m,6H), 3.51 -3.62 (m,3H),6.21-6.2 (m,lH), 6.19-6.29 (m,lH),

6.88-7.1 (m,lH), 8.47(s,3H), 10.98 (s,lH)

IH NMR (400 MHz, DMS0-d6) δ 0.82-0.90 (m, IH), 0.95-1.05 (m, IH), 1.10-1.35 (m, 3H), 1.68-1.88 (m, 4H), 1.89-2.0 (m, 3H),

300

2.05-2.15 (m, 2H), 2.95-3.05 (m, 2H), 3.10- 3.25 (m, 2H), 3.85-4.10 (m, 3H), 6.02-6.90 (m, 2H), 7.93 (brs, 3H)

IH NMR (400 MHz, DMSO-d6) δ 0.84 ((t, J =6.89Hz, 2H), 1.72-1.85 (m, 2H), 1.88-2.0

301 (m, 2H), 3.10-3.13 (m, 2H), 3.28-3.42 (m, 7H), 3.6-3.75 (m, 4H), 6.62 (d, J=22.87Hz,

2H), 8.32 (s, 3H), 8.76 (s, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.92 (brs, 4H), 3.02 (brs, 3H), 3.15-3.35 (m, 9H), 6.20-

302 6.47 (m, 2H), 6.80-7.01 (m, IH), 7.02-7.20

(m, IH), 7.21-7.42 (m, IH), 8.25 (s, 4H), 9.45 (s, 2H), 10.01 (brs, IH)

IH NMR (400 MHz, DMSO-d6) δ 1.92 (s, 2H), 2.10 (s, 2H), 3.13-3.19 (m, 2H), 3.29-

303 3.37 (m, 7H), 3.61 (s, 5H), 6.28-6.37 (m, 2H), 6.60 (s, 2H), 6.85-6.87 (m, IH), 7.94

(brs, IH), 8.22 (s, 3H), 10.89 (brs, IH)

IH), 12.65 (brs, IH)

2H), 8.40 (s, 3H), 11.00 (brs, IH)

IH), 10.87 (brs, IH) Cmpd# Structure NMR data

1H NMR (400 MHz, DMSO-d6) δ 1.33-1.34 (m, 2H), 1.52-1.62 (m, 4H), 1.93 (d, J

=10.2Hz, 2H), 2.11 (d, J=11.8Hz, 2H), 3.14-

327 3.16 (m, 2H), 3.30 (s, 4H), 3.38 (s, 1H), 3.62

(d, J=8.9Hz, 3H), 6.57 (d, J=6.2Hz, 3H), 8.33 (s, 3H), 10.94 (brs, 1H)

1H NMR (400 MHz, DMSO-d6): δ 9.52 (brs,

328 1H), 7.99 (brs, 1H), 7.60-6.80 (m, 3H), 3.20 (brs, 3H), 2.1 (brs, 3H), 1.93 (brs, 4H)

1H NMR (400 MHz, DMSO-d6): δ 10.90- 10.81 (m, 1H), 8.41(brs, 3H), 8.06 (s, 1H), 7.18-7.05 (m, 2H), 6.86 (d, J_= 8.53 Hz, 1H),

329

6.32 (s, 1H), 6.13 (s, 1H), 3.70-3.58 (m, 3H), 3.37-3.31 (m, 4H), 3.12-3.09 (m, 2H), 2.14- 2.09 (m, 5H), 1.94-1.88 (m, 2H)

1H NMR (400 MHz, DMSO-d6) δ 1.21 (s, 2H), 1.89-1.93 (m, 4H), 2.09 (s, 2H), 3.07 (s,

330 4H), 3.14-3.17 (m, 4H), 3.27 (s, 4H), 3.36 (s,

2H), 3.56 (s, 8H), 3.62-3.68 (m, 3H), 6.56 (brs, 2H), 9.62 (brs, 1H)

1H NMR (400 MHz, DMSO-d6): δ 10.23 (brs, 1H), 8.52 (brs, 1H), 8.08 (brs, 6H), 7.18- 7.15 (m, 2H), 6.95 (brs, 1H), 6.65 (s, 1H),

331

6.58 (s, 1H), 3.65-3.63 (m, 4H), 3.28 (brs, 2H) 3.11 (brs, 2H), 2.98 (s, 4H), 2.84 (s, 2H), 2.69-2.65 (m, 4H), 2.20-1.90 (m, 4H) Cmpd# Structure NMR data

1H NMR (400 MHz, DMSO-d6) δ 1.04-1.22 (m, 2H), 1.32-1.48 (m, 6H), 1.75-1.93 (m, 3H), 2.10-2.13 (m, 3H), 2.51-2.60 (m, 1H)

337 xx ^N xi 3.13-3.17 (m, 3H), 3.28-3.29 (m, 6H), 3.37- 3.42 (m, 1H), 4.40-4.42 (m, 2H), 6.65-6.67 (m, 2H), 6.94-7.07 (m, 2H), 8.34-8.38 (m, 3H), 10.89 (brs, 1H)

1H NMR (400 MHz, DMSO-d6) δ 1.11 -1.24 (m, 1H), 1.74-2.11 (m, 3H), 3.05-3.17 (m,

338 3H), 3.31 -3.49 (m, 9H), 3.56-3.72 (m, 4H),

6.93 (s, 1H), 7.08-7.34 (m, 3H), 7.45 (s, 1H), 8.35 (s, 4H), 10.98 (brs, 1H)

1H NMR (400 MHz, DMSO-d6) δ 1.03 (t, J =6.9Hz, 3H), 1.92-1.98 (m, 2H), 2.12 (d, J =13.2Hz, 2H), 3.00 (s, 3H), 3.11-3.18 (m, 2H),

339 3.30 (s, 3H), 3.37-3.48 (m, 3H), 3.55-3.68 (m, tH), 6.63 (d, J=22.1Hz, 2H), 7.14 (d, J=7.4Hz, 1H), 7.36-7.47 (m, 1H), 8.30 (s, 3H), 10.85 (s,

1H), 12.61 (brs, 1H)

LH NMR (400 MHz, DMSO-d6) δ 1.44 (s, 4H), 1.11-2.13 (m, 5H), 3.14 (t, J=11.9Hz, 2H), 3.29-

340 3.36 (m, 4H), 3.65 (s, 8H), 7.21 (br s, 1H), 8.40

(s, 3H), 10.96 (s, 1H)

H NMR (400 MHz, DMSO-d6) δ 1.11 -1.21 m, 8H), 1.54 (s, 1H), 1.74-2.10 (m, 2H), 3.03-

341 .19 (m, 2H), 3.31 -3.48 (m, 6H), 3.64-3.68 (m,

H), 6.52-6.75 (m, 2H), 7.07-7.65 (m, 2H), 8.34 s, 3H), 10.81-10.97 (m, 1H) Cmpd# Structure NMR data

¾ NMR (400 MHz, DMSO-d ₆ ) δ 1.4-1.49 (m, 2H), 1.6-1.7(m, 2H), 1.7-1.8(m,2H), 1.9-2.0(m 4H), 2.1(d, J=13.1Hz, 2H), 2.8-2.9(m, 1H),

354 3.1-3.2(m, 2H), 3.27-3.33 (m, 4H), 3.6 (d, J

=11.8Hz, 3H), 5.6(brs, 1H), 6.6(s, 1H), 6.7(s, 1H), 6.8(s, 1H), 6.9(d, J=8.4Hz, 2H), 8.2(br, 4H), 10.94(brs, 1H).

¾ NMR (400 MHz, DMSO-d ₆ ) δ 1.1 -1.2 (m, 2H), 1.9-2.1 (m, 2H), 3.2(br s, 8H), 6.5(br

355

s,lH), 6.8-6.9 (m, 2H), 7.5 (s, 1H), 8.1-8.3 (m, 3H), 8.8 (br s, 1H), 9.3 (br s, 1H).

¾ NMR (400 MHz, DMSO-d ₆ ) δ 1.9-2.0 (m, 6H), 2.1-2.2 (m, 2H), 3.1-3.2(m, 5H), 3.3-3.4

356 (m, 5H), 3.6 (d, J= 11.28 Hz ,3H), 6.1 (br, 1H), 6.3 (s, 1H ), 6.5 (s, 2H), 6.6 (s ,1H), 6.7- 6.8 (m, 1H), 8.1 (br, 1H), 8.3 (br s, 3H).

¾ NMR (400 MHz, DMSO-d ₆ ) δ 0.8 (d, J = 6.44, 6H), 1.1-1.3 (m, 2H), 1.6-1.7 (m, 1H),

357 1.9-2.0 (m, 3H),2.0-2.1 (m, 3H), 3.3 (s, 4H),

3.4-3.5 (m, 6H ), 6.5 (s, 1H), 6.6 (s ,1H), 6.7- 6.9 (m, 2H), 8.3 (br s, 3H).

¾ NMR Γ400 MHz. DMSO-dfi) 51.8- 1.9 Cm. 2H). 2.1-2.19 Cm. 2H). 2.4-2.5im. 2H). 2.5- 2.55im. 1H). 3.19-3.25 (m. 10H). 3.5-3.7 Cm.

358

2H). 6.14iS. 1H). 6.5id. J= 6.8Hz. 2H).

7. lis. 3HV 8.23ibrs. 3H). 8.4ibrs. 1H).

9.2ibrs. 2HY Cmpd# Structure NMR data

¾ NMR (400 MHz, DMSO-d ₆ ) δ 1.22 (s, 9H), 1.3 (s, 18H), 1.5-1.6(m, 3H), 1.81-1.9(m,lH), 1.97 (t, J= 7.4 Hz,3H), 2.17-2.2(m, 2H), 2.31- 2.39(m ,1H), 2.43(t, J=6.72 Hz,2H), 2.6-2.7 (n

359 2H), 2.84-2.89 (m, 2H), 3.29-3.43(m, 1H),

3.56(t, J=6.5Hz, 2H), 5.42(br, 1H), 6.34(s, 1H 6.84(s, 1H), 6.96(s, 1H), 7.46(d, J=8.3Hz, 1H) 7.51(s, 1H), 7.716(d, J= 8.32, 1H).

Anti-infective activity of the synthesised compounds

The compounds as disclosed by the present application have anti-infective activity.

Initial minimal inhibitory concentration (MIC) tests were made on two bacterial strains:

- Escherichia coli (ATCC25922)

- Staphylococcus aureus (ATCC25923).

The results of these tests are shown in Table 15.

The MIC of selected compounds was determined against a number of further strains:

Enterococcus faecalis (ATCC29212)

Pseudomonas aeruginosa (ATCC27853)

Staphylococcus aureus subsp. aureus (ATCC29213)

Klebsiella pneumoniae subsp. pneumoniae (ATCC13883) Streptococcus pneumoniae (ATCC33400) Haemophilus influenzae (ATCC49766) Neisseria meningitidis (ATCC 13077) Listeria monocytogenes (ATCC15313) Legionella pneumophila subsp. pneumophila (ATCC33152) Mycobacterium bovis BCG (ATCC 19210)

The results of these tests are shown in Table 16.

Minimal Inhibitory Concentration βΤΙϋ)

MIC values were determined using the standard broth microdilution procedure based on the guidelines by the Clinical and Laboratory Standards Institute (CLSI). Briefly, the compounds were dissolved in DMSO to 10 mM. They were diluted in cation-adjusted Mueller-Hinton broth (CAMHB) to four times the highest concentration tested. A serial 2-fold dilution in CAMHB was done in microdilution plates. The inoculum of bacterial strain to be tested was prepared by making a suspension of colonies from an 18 to 24 hours old plate in CAMHB. The inoculum was diluted so that, after inoculation, each well contained approximately 5 x 10 ⁵ CFU/mL. To a volume of 50 μΐ compound in CAMHB an equal volume of inoculum was added. The tray was sealed in a plastic bag and incubated at 35°C for 16 to 20 hours. To aid in the detection of growth the dye resazurin was added to a final concentration 0.001% and incubated at room temperature for 1 h. Reduction of resazurin, and therefore bacterial growth, was seen as a change from blue to pink. The MIC is the lowest concentration of compound that completely inhibits growth of the organism.

The method used is described in detail in: Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard— Ninth Edition. CLSI document M07-A9. Wayne, PA: Clinical and Laboratory Standards Institute; 2012. Inhibition of bacterial RNaseP activity.

The assay is based on how much the cleavage of the substrate pATSerUG by E. coli RNase P RNA, Ml RNA, is inhibited by the compound. The substrate pATSerUG is a 45 nt long model substrate that maintains T-stem/loop structure of the tRNA ^{s r} precursor. It was purchased from Dharmacon/GE Healthcare, and labelled with ³² P at the 5' end with [γ- ³² Ρ]ΑΤΡ according to standard procedures, and purified by electrophoresis on a denaturing polyacrylamide gel. The Ml RNA was generated by T7 in vitro transcription using a PCR product with the Ml RNA gene as template.

The compound to be tested was dissolved in assay buffer (see below). Assay buffer was added to a theoretical concentration of up to 10 mM. After vortexing and incubation at room temperature for 30 minutes the undissolved compound was removed by centrifugation (17,000xg 10 min). The concentration of compound in the supernatant was determined spectroscopically by measuring the absorbance at a wavelength where the compound had an absorbance maximum. The calibration curve was made from known concentrations of the compound dissolved in DMSO.

The cleavage reaction was performed in assay buffer (50 mM Tris-HCl, pH 7.9, 1 mMNELtCl, 10 mM MgCl ₂ , 5% PEG6000, 10 mM spermidine).

Ml RNA was diluted to 10 times the concentration to be used in assay buffer and preincubated at 37°C for 10 min to allow proper folding. The final concentration of Ml RNA was determined for each batch of enzyme, and was the concentration that gave approximately 50% cleavage of the substrate in a 10 min reaction. The folded Ml RNA was mixed with the compound to be tested in a total volume of 9 μΐ and incubated for an additional 10 min at 37°C. The substrate was preheated separately for 5 min at 37°C. The reaction was started by the addition of Ι μΐ substrate to the Ml RNA-compound mixture. After 10 min incubation at 37°C the reaction was stopped by the addition of 20 μΐ stop solution (10 M urea, 100 mM EDTA, 0,05% bromophenol blue, 0,05% xylene cyanol). The reactions were then heated to 95°C for 3 min, chilled on ice, then resolved on 20% polyacrylamide/7 M urea/TBE gels and detected using a Phosphoimager. The signals were quantitated using the softwares QuantityOne or ImageLab.

Initial inhibition of RNase P activity

To test if any inhibition could be detected for the compound an initial inhibition of RNase P activity was determined. The maximum amount of compound was used, ie. 8 μΐ of the supernatant from freshly dissolved compound in assay buffer in a 10 μΐ cleavage reaction. The degree of inhibition was judged from the normalised cleavage (the ratio between cleavage with compound divided by cleavage without compound). If this ratio was <0,5, the IC50 value was determined. IC50 determination.

About 8 different concentrations, generally ranging from maximum concentration for the compound down to 8000 times diluted, were tested for cleavage. The IC50 values and Hill slopes were calculated using the software GraphPad Prism. The determined IC50 values are listed in Table 15.

Table 15: RNase P inhibition and Antibacterial Efficacy Results

NA: Not available NI: No inhibition

Table 16: MIC of selected compounds against a range of bacteria

faecalis pneumoniae

Strain: ATCC ATCC ATCC JW5503 ATCC

ATCC 11758 29212 700221 25922 43816

Cmpd # MIC MIC MIC MIC MIC

MIC ( g/ml) ^g/ml) ^g/ml) ^g/ml) ^g/ml)

44 16 8 >128 8 >128

92 2 0,5 >128 >128 >128

1050 32 16 32 16 32

111 4 2 64 4 >128

123 2 2 >128 4 >128

133 8 4 32 8 32

140 4 4 32 8 8

150 2 2 16 4 16

186 4 4 16 4 128

197 2 2 64 4 >128

24239 4 2 64 64 128

245 2 1 8 8 32

255 16 32 64 16 128

274 2 1 4 8 4 4

284 2 2 4 4 4 4

300 1 1 4 2 8 4

301 2 2 4 2 4 8

302 8 4 16 16 16 16

305 1 1 4 4 4 2

306 4 2 4 2 4 8

307 2 2 4 2 4 8

Organism: aureus _* S aureus _* S aureus S. S. aureus

+ 50% HS MRSA pneumoniae USA300 M. fortuitum

MRSA

Strain: ATCC ATCC ATCC ATCC BAA ATCC 29213 29213 33591 49619 1717 110

Cmpd # MIC MIC MIC MIC MIC MIC

^g/ml) ^g/ml) ^g/ml) ^g/ml) ^g/ml)

44 4 >128 64 16

92 0,5 128 2 8

1050 16 128 32 32

111 2 128 4 8

123 2 64 8 8

133 4 128 4 32

140 4 128 4 32

150 2 64 2 8

186 4 64 8 32

197 2 128 16 4

242 2 128 2 16

245 2 128 2 16

255 8 64 16 64

274 1 16 1 4 1 4

284 2 32 2 8 2 8

300 1 16 1 4 1 2

301 4 16 2 4 2 4

302 8 64 8 64 8 8

305 1 32 1 2 1 4

306 2 8 2 4 2 4

307 2 32 2 8 2 4