BRONCHORELAXING AGENTS BASED ON INDOL AND ISOQUINOLINE

DERIVATIVES

FIELD OF THE INVENTION The present invention relates to novel bronchorelaxing agents, pharmaceutical compositions comprising such agents, and a method of treating or allevating conditions accompanied by bronchoconstriction.

BACKGROUND OF THE INVENTION

Airway obstruction, accompanied by an increase in the contractile state of the bronchial smooth muscle, is prominent in a number of diseases of the respiratory apparatus, in particular asthma, chronic obstructive pulmonary disease (which comprises chronic bronchitis and emphysema), bronchiectasis, cystic fibrosis, bronchiolitis and bronchopulmonary dysplasia. Bronchoconstriction may be caused by a number of factors that affect the bronchi and other parts of the respiratory apparatus independent of each other or in combination. The available means for treating or preventing bronchoconstriction are insufficient in many respects. Thus new agents that exert a relaxing effect on constricted bronchi are much in need.

OBJECTS OF THE INVENTION

It is an object of the present invention to provide an agent for treating or preventing bronchoconstriction and for use in treating diseases such as asthma, in which bronchoconstriction is prominent.

It is another object of the present invention to provide a pharmaceutical composition comprising said agent. Still another object of the present invention is to provide a method for treating or preventing bronchoconstriction by administration of such agent to a person in need.

Further objects of the invention will become apparent from the following summary of the invention, the description of preferred embodiments thereof, and the appended claims.

SUMMARY OF THE INVENTION According to the present invention is disclosed an agent for treating or preventing bronchoconstriction in form of a chemical compound of the general formula (I) including its pharmaceutically acceptable acid addition salts

(I)

wherein R ₁ , R _2, R ₃ , R ₄ are, independent of each other:

(a) H ₅ C ₁ -C _S aIlCyI;

(b) halogen;

5 (c) NR ₅ R ₆ , wherein R ₅ and R ₆ are, independent of each other, H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, SO ₂ R ₇ , wherein R ₇ is C ₁ -C ₆ alkyl, CF ₃ , aryl or substituted aryl;

(d) CN;

(e) COR ₈ , wherein R ₈ is H, OH, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy;

(f) SO ₂ R ₉ wherein R ₉ is OR ₁ O, wherein R ₁₀ is H or C ₁ -C ₆ alkyl or NR ₁₁ R ₁₂ , 0 wherein R ₁₁ and R ₁₂ are, independent of each other, H, C ₁ -C ₆ alkyl;

(g) OR ₁₃ , wherein R ₁₃ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, C ₁ -C ₈ carboxy, C ₁ -C ₈ carbamoyl; wherein, if R ₂ and R ₃ both are OR ₁₃ , R ₁₃ may additionally be CHR ₁₄ or CO 5 shared by R ₂ and R ₃ , R ₁₄ being selected from hydrogen and C ₁ -C ₆ alkyl;

X is O or S;

A is H, C ₁ -C ₆ alkyl, which may be substutited by aryl or substituted aryl; 0

B is Cj-C ₁₈ alkyl, which may be mono- or di-unsaturated and/or substituted by alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, wherein, independent of each other, said C ₁ -C ₁₈ alkyl and said alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl substituting the C ₁ -C ₁₈ alkyl may be further 5 substituted by one to three of F, Cl, Br;

M is zero or one, with the proviso that no more than three OfR ₁ , R ₂ , R ₃ , R ₄ are H and with the further O proviso that

(i) IfR ₁ and R ₄ are H, m is 1 , X is S or O, A is any of allyl, 2-

(4-chlorophenyl)ethyl, methyl, 2-phenyletyl, octyl, substituted or unsubstituted 2-imidazolyl, 2- thienyl and B is H or methyl, R ₂ and R ₃ are not both OCH ₃ or both OH or 5 OH and OCH ₃ ;

(ii) if R ₃ is NH ₂ and m is 1 and X is O, A and B are not both ethyl;

(iii) if R ₁ is Cl, m is O and X is O, A and B are not 2-chloroethyl;

(iv) if R ₁ is OCH ₃ , m is O and X is S. A and B are not 2- O phenethyl;

(v) if R ₁ and R ₄ are H, m is O, X is S, A is 2-(4-chlorophenyl)- ethyl and B is H, R ₂ and R ₃ are not OH or OH and 0CH _3; (vi) if R ₁ and R ₂ are H, m is 1, X is S, A is 2-(4- chlorophenyl)ethyl and B is H, R ₃ and R ₄ are not OH. 5

The pharmaceutically acceptable addition salts as mentioned hereabove comprise the therapeutically active non-toxic addition salt forms which the compounds of the general formula (I) are able to form. They can conveniently be obtained by treating the base form with appropriate inorganic, such as, for instance, hydrochloric acid, O hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with appropriate organic acids, such as, for instance, acetic, propanoic, methanesulfonic,

benzenesulfonic, lactic, malic, citric, tartaric, succinic, maleic acid and the like. The term acid addition salt also comprises the hydrates and solvent addition forms, such as hydrates and alcoholates, which the compounds of the general formula (I) are able to form.

5 According to a first preferred aspect of the invention, in the compound of the general formula (I), R ₃ is OH and R ₁ , R ₂ , R ₄ are, independent of each other H; C ₁ -C ₆ alkyl; halogen; NR ₅ R ₆ , wherein R ₅ and R ₆ are, independent of each other, H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, SO ₂ R ₇ , wherein R ₇ is C ₁ -C ₆ alkyl, CF ₃ , aryl or substituted aryl; CN; COR ₈ , wherein R ₈ is H, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy; SO ₂ R ₉ , wherein R ₉ is OR ₁₀ , wherein R ₁₀ is H or C ₁ - 0 C ₆ alkyl, C i -C ₆ alkyl or NR ₁ iR ₁₂ , wherein R ₁₁ and R ₁₂ are, independent of each other, H, C ₁ -C ₆ alkyl; OR ₁₃ , wherein R ₁₃ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, C ₁ -C ₈ carboxy or C ₁ -Cg carbamoyl; X is O; M is 1.

According to a second preferred aspect of the invention in the compound of 5 the first preferred aspect R ₃ is OH and R ₁ , R ₂ , R ₄ are, independent of each other H; C ₁ -C ₆ alkyl; halogen; NR ₅ R ₆ , wherein R ₅ and R ₆ are, independent of each other, H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl; SO ₂ R ₇ , wherein R ₇ is C ₁ -C ₆ alkyl, CF ₃ , aryl or substituted aryl; CN; COR ₈ , wherein R ₈ is H, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy; SO ₂ R ₉ wherein R ₉ is OR _1O , wherein R ₁₀ is H or C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl or NR ₁₁ R ₁₂ , wherein R ₁₁ and R ₁₂ are, independent of each 0 other, H, C ₁ -C ₆ alkyl.

According to a third preferred aspect of the invention in the compound of the first preferred aspect R ₃ is OH and R ₁ , R ₂ , R ₄ are, independent of each other H; C ₁ -C ₆ alkyl; halogen; CN; COR ₈ , wherein R ₈ is H, C ₁ -C ₆ alkyl or Ci-C ₆ alkoxy; OR ₁₃ , wherein 5 R ₁₃ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, C ₁ -C ₈ carboxy or C ₁ -C ₈ carbamoyl.

According to a fourth preferred aspect of the invention in the compound of the third preferred aspect most preferred in consideration of the foregoing restrictions R ₃ is OH and R ₁ , R ₂ , R ₄ are, independent of each other H; C ₁ -C ₆ alkyl; halogen; CN; OR ₁₃ , O wherein R ₁₃ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, C ₁ -C ₈ carboxy or C ₁ -C ₈ carbamoyl.

According to a fifth preferred aspect of the invention in the compound of the second preferred aspect R ₃ is OH and R ₁ , R ₂ , R ₄ are, independent of each other H; C ₁ -C ₆ alkyl; halogen; NR ₅ R ₆ , wherein R ₅ and R ₆ are, independent of each other, H, C ₁ -C ₆ alkyl, 5 C ₂ -C ₆ acyl; SO ₂ R ₇ , wherein R ₇ is C ₁ -C ₆ alkyl, CF ₃ , aryl or substituted aryl; CN; COR ₈ , wherein R ₈ is H, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy; SO ₂ R ₉ , wherein R ₉ is OR ₁₀ , wherein R ₁₀ is H or C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl; OR ₁₃ , wherein R ₁₃ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, C ₁ -C ₈ carboxy or Ci-C ₈ carbamoyl. O Particularly preferred are the compounds:

According to a sixth preferred aspect of the invention in the compound of the O general formula (I) R ₁ -R ₄ are, independent of each other, H; C ₁ -C ₆ alkyl; halogen; NR ₅ R ₆ , wherein R ₅ and R ₆ are, independent of each other, H, Ci-C ₆ alkyl, C ₂ -C ₆ acyl; SO ₂ R ₇ ,

wherein R ₇ is C ₁ -C ₆ alkyl, CF ₃ , aryl or substituted aryl; CN; COR ₈ , wherein R ₈ is H, OH, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy; SO ₂ R ₉ , wherein R ₉ is OR ₁₀ , wherein R ₁₀ is H or C ₁ -C ₆ alkyl OrNR ₁₁ R ₁₂ , wherein R ₁₁ and R ₁₂ are, independent of each other, H ₃ C ₁ -C ₆ alkyl; X is S; M is 1 ; with the proviso that neither R ₂ and R ₃ are both H nor R ₅ and R ₆ are both H.

According to a seventh preferred aspect of the invention in the compound of the sixth preferred aspect R ₁ and R ₄ are H; R ₂ and R ₃ are, independent of each other, H; C ₁ -C ₆ alkyl; halogen; NRsR ₆ , wherein R ₅ and R ₆ are, independent of each other, H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl; SO ₂ R ₇ , wherein R ₇ is C ₁ -C ₆ alkyl, CF ₃ , aryl or substituted aryl; CN; COR ₈ , wherein R ₈ is H, OH, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy; SO ₂ R ₉ , wherein R ₉ is OR ₁₀ , wherein R ₁₀ is H or C ₁ -C ₆ alkyl or NR ₁₁ R ₁₂ , wherein Rn and R ₁₂ are, independent of each other, H, C ₁ -C ₆ alkyl.

According to an eighth preferred aspect of the invention in the compound of the seventh preferred aspect R ₂ and R ₃ are both C ₁ -C ₆ alkyl; halogen; NR ₅ R ₆ , wherein R ₅ and R ₆ are, independent of each other, H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl; SO ₂ R ₇ , wherein R ₇ is C ₁ -C ₆ alkyl, CF ₃ , aryl or substituted aryl; CN; COR ₈ , wherein R ₈ is H, OH, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy; SO ₂ R ₉ , wherein R ₉ is OR ₁₀ , wherein R ₁₀ is H or C ₁ -C ₆ alkyl or NR ₁₁ R ₁₂ , wherein R ₁₁ and R ₁₂ are, independent of each other, H, C ₁ -C ₆ alkyl.

Particularly preferred are the compounds:

According to a nineth preferred aspect of the invention in the compound of the general formula (I) R ₁ and R ₄ are halogen; R ₂ and R ₃ are, independent of each other, OR ₁₃ , wherein R ₁₃ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, C ₁ -C ₈ carboxy, C ₁ -C ₈ carbamoyl; wherein, if R ₂ and R ₃ both are OR ₁₃ , R ₁₃ may additionally be CHRi ₄ or CO shared by R ₂ and R ₃ , R ₁₄ being selected from hydrogen, C ₁ -C ₆ alkyl; X is S; M is 1.

According to a tenth preferred aspect of the invention in the compound of the nineth preferred aspect R ₂ and R ₃ are, independent ocf each other, OR ₁₃ , wherein R ₁₃ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ acyl, C ₁ -C ₈ carboxy; wherein, if R ₂ and R ₃ both are OR ₁₃ , R ₁₃

may additionally be CHR ₁₄ or CO shared by R ₂ and R ₃ , R ₁₄ being selected from hydrogen, C ₁ -C ₆ alkyl.

Particularly preferred is the compound Res-13-61 :

According to an eleventh preferred aspect of the invention in the compound of the general formula (I) R ₁ and R ₄ are, independent of each other, H or halogen; R ₂ and R ₃ are OH; X is S; M is 1; A is H; B is 2-(2, 3 or 4-pyridinium-N-oxide)ethyl or an acid addition salt of 2-(2, 3 or 4-N-alkylpyridinium)ethyl, alkyl being selected from C ₁ -C ₆ alkyl.

Particularly preferred is the compound Res-13-55

According to a twelvth preferred aspect of the invention in the compound of the general formula (I) R ₃ and R ₄ are H; R ₁ and R ₂ are H or OH, with the proviso that, if R ₁ is H R ₂ is OH, and if R ₁ is OH, R ₂ is H; X is S; A is H; B is 2-(4-chloro-phenyl)ethyl; M is O:

H

Particularly preferred are the compounds:

Res-14-84

The term "C ₁ -C ₆ alkyl" comprises straight and branched chain alkyl, such as methyl, ethyl, propyl, isoproyl, butyl, isobutyl, t-butyl, pentyl, 2-methylbutyl, hexyl, 2- methylpentyl.

The term "C ₂ -C ₆ acyl" comprises straight and branched chain acyl, such as acetyl, propionyl, butyryl, iso-butyryl.

The term "halogen" comprises F, Cl, Br, I.

The compounds of the invention have been tested for their bronchoconstriction-inhibiting or bronchorelaxing effect in a model comprising a human bronchus preparation. The model is described in detail in the Preferred Embodiments section. Particularly preferred compounds according to the invention are those which exhibit in this model a bronchorelaxing effect which is about the same or even better than that of capsazepine on a weight/weight basis.

Most preferred compounds according to the invention are those which exhibit in this model a bronchorelaxing effect which is superior to that of capsazepine on a weight/weight basis The compounds of the present invention and their pharmaceutically acceptable acid addition salts can be used in the treatment of diseases in which the

constriction of the bronchi is of importance, such as asthma. The present compounds may block bronchoconstriction agonist-induced contractions of bronchial tissues.

The compounds of the invention can therefore be used as medicines against above-mentioned diseases or in their prevention. Said use as a medicine or method of treatment comprises the systemic administration to patients of an amount effective to combat bronchoconstriction.

The compounds of the invention can be formulated into various pharmaceutical forms for administration purposes. Said pharmaceutical forms or compositions are deemed novel and consequently constitute another aspect of the present invention. Also the preparation of said compositions constitutes a further aspect of the present invention. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, including in acid addition salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are desirably in unitary dosage form suitable, preferably, for administration orally, rectally, percutaneously, or by parenteral injection. Particularly preferred is administration by inhalation.

For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs and solutions: or solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like in the case of powders, pills, capsules and tablets. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will usually comprise sterile water, at least in large part, though other ingredients, for example to aid solubility, may be included. Injectable solutions, for example, may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. In the compositions suitable for percutaneous administration, the carrier option-ally comprises a penetration enhancing agent and/or a suitable wetting agent, optionally combined with suitable additives of any nature in minor proportions, which additives do not introduce a significant deleterious effect on the skin. Said additives may facilitate the administration to the skin and/or may be helpful for preparing the desired compositions. These compositions may be administered in various ways, e.g., as a transdermal patch, as a spot-on or as an ointment.

Acid addition salts of the compound of general formula (I) due to their increased water solubility over the corresponding base form, are obviously more suitable in the preparation of aqueous compositions. It is especially advantageous to formulate the aforementioned pharmaceutical compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used in the specification and claims herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingre-dient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such dosage unit forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, injectable solutions or suspensions, teaspoonfuls, tablespoonfuls and the like, and segregated multiples thereof. Administration by inhalation will allow a high proportion of

the delivered dose to reach the site of action, that is, the bronchi and the lung in general. Inhalation may be by the oral or the nasal route. Conventional pulmonary applicators may be employed, such as pressurized spray containers containers suitable propellants for aerosols and powder spray devices for preparations in form of fine powders. Pharmaceutical compositions suitable for administration by the inhalation route are known in the art. The compound is dissolved in a suitable vehicle or employed as a fine powder, such as a micronized powder of a particle size from about 2 μm to about 20 μm. An indicated daily dose for administration by inhalation will be 10 times and more lower than the oral dose. Satisfactory doses, preferably metered by using a device capable of metering, or by single doses of predetermined size, can easily be determined by experimentation.

In view of the usefulness of the compounds of the invention in the treatment of diseases in which bronchoconstriction is prominent, it is evident that the present invention provides a method of treating warm-blooded animals suffering from such diseases, said method comprising the systemic administration of a pharmaceutically effective amount of a compound of formula (I) or a pharmaceutically acceptable acid addition salt thereof in admixture with a pharmaceutical carrier. Those of skill in the treatment of diseases in which bronchoconstriction is an important factor could easily determine the effective amount. In general it is contemplated that an effective amount would be from 0.01 mg/kg to 4 mg/kg body weight, preferably from 0.04 mg/kg to 2 mg/kg body weight.

The exact dosage and frequency of administration depends on the particular compound of formula (I) used, the particular condition being treated, the severity of the condition being treated, the age, weight and general physical condition of the particular patient as well as other medication the individual may be taking, as is well known to those skilled in the art. Furthermore, it is evident that said effective daily amount may be lowered or increased depending on the response of the treated subject and/or depending on the evaluation of the physician prescribing the compounds of the instant invention. The effective daily amount ranges mentioned hereinabove are therefore guidelines only and are not intended to limit the scope or use of the invention.

According to a preferred aspect of the invention the compounds of the invention can be combined with an anti-asthmatic, in particular an anti-asthmatic selected from β ₂ -agonist, anticholinergic, corticosteroid, and calcium antagonist, for the treatment of asthma and related conditions. Also disclosed is a pharmaceutical composition comprising a bronchorelaxing amount of a compound of the invention in combination with a pharmacologically airway-effective amount of β ₂ -agonist, anticholinergic, corticosteroid, calcium channel blocker or a mixture thereof, and a pharmaceutically acceptable carrier, and its administration to a patient suffering from asthma or a related condition characterized by bronchoconstriction.

It is preferred for the β ₂ -agonist to be selected from: adrenaline; albuterol; amiterol; bambuterol; bitolterol; buphenine; broxaterol; carbuterol; cimaterol; clenbuterol; clorprenaline; colterol; denopamine; dioxethedrine; dioxifedrine; dopexamine; doxaminol; dobutamine; etanterol; ephedrine; epinephrine; adrenaline; eprozinol; etafedrine; ethylnorepinephrine; fenoterol; berotec; dosberotec; partusisten; flerobuterol; formoterol; eformoterol; r,r-formoterol; hexoprenaline; ibopamine; isoeharine; ibuterol; imoxiterol; isoxsuprine; ibuterol; isoprenolol; isoproterenol; levalbuterol; r-form of albuterol; levosalbutamol; levisoprenaline; 1-form of isoprenaline; mabuterol; meluadrine;

mesuprine; metaterol; metaproterenol; methoxyphenamine; nardeterol; oxyfedrine; orciprenalin; picumeterol; pirbuterol; prenalterol; procaterol; protokylol; quinprenaline; reproterol; rimiterol; ritodrine; salbutamol; albuterol; salmeterol; soterenol; sulphonterol; ta-2005; terbutaline; tretoquinol; tulobuterol; xamoterol; zilpaterol; ar-c68397aa; 4- hydroxy-7-[2-[2-[3-phenylethoxypropane-l-sulfonyl]ethylamino ]ethyl]-3h-benzothiazol- 2-one hydrochloride; chf-1035; rac-5,6~diiso-butyryloxy-2-methylamino- 1,2,3,4- tetrahydronaphthalene hydrochloride; hoku-81; l-(2-chloro-4-hydroxyphenyl)-2-tert- butylaminoethanol; ibuterol; 1 -(3 ,5-dihydroxyphenyl)-2-(tert-butylamino)ethanol diisobutyrate ester; meluadrine; 4-(2-tert-butylamino-l-hydroxyethyl)-3-chlorophenol; ta- 2005; 8-hydroxy-5-[(lr)-l-hydroxy-2-[n-[(lr)-2-(p-methoxyphenyl)-l -methylethyl]- amino]ethyl]carbostyril hydrochloride; tiaramide; 5-chloro-3~[4-(2-hydroxyethyl)-l- piperazinyl]carbonyl-methyl-2-benzo-thiazolinone; trimetoquinol; (1 ,2,3,4-tetrahydro-l - ((3,4,5-trimethoxyphenyl)methyl)-6,7-isoquinolinediol); desformoterol; ((r,r) or (s,s)-3- amino-4-hydroxy-.alpha.-(((2-(4-methoxy-phenyl)-l-methylethy l)amino)methyl)- benzenemethanol ; 4-hydroxy-7- [2- { [2- { [3 ~(2-phenylethoxy)propyl] sulphonyl } -ethyl] - amino}-ethyl]-2(3h)-benzothiazolone; l-(2-fluoro-4-hydroxyphenyl)-2-[4-(l- benzimidazolyl)-2-methyl-2-butylamino]-ethanol; l-[3-(4-methoxybenzyl-amino)-4- hydroxyphenyl]-2-[4-(l-benzimidazolyl)-2-methyl-2-butylamino ]ethanol; l-[2h-5- hydroxy-3-oxo-4h-l,4-benzoxazin-8-yl]-2-[3-(4-n,n-dimethyl-a minophenyl)-2-methyl-2- propylaminojethanol; l-[2h-5-hydroxy-3-oxo-4h-l,4-benzoxazin-8-yl]-2-[3-(4~ methoxyphenyl)-2-methyl-2-propylamino]ethanol; 1 -[2h-5-hydroxy-3-oxo-4h- 1 ,4- benzoxazin-8-yl]-2-[3-(4-n-butyloxy-phenyl)-2-methyl-2-propy lamino]ethanol; l-[2h-5- hydroxy-3 -oxo-4h- 1 ,4-benzoxazin- 8-yl] -2- { 4- [3 -(4-methoxyphenyl)- 1 ,2,4-triazol-3 -yl] -2- methyl-2-butylamino}ethanol; 5-hydroxy-8-(l-hydroxy-2~isopropylamino-butyl)-2h-l,4- benzoxazin-3-(4h)-one; 1 -(4-ammo~3-chloro-5-trifluoromethylphenyl)-2-tert.- butylamino)ethanol; l-(4-ethoxycarbonylamino-3-cyano-5-fluorophenyl)-2-(tert.- butylamino)ethanol.

It is preferred for the anticholinergic to be selected from: adiphenine, alverine, ambutonium, bromide, aminopentamide, amixetrine, amprotropine phosphate, anisotropine methylbromide, apoatropine, atropine, atropine, n-oxide, benactyzine, benapryzine, benzetimide, benzilonium, benzilonium bromide, benztropine mesylate, bevonium methyl, sulfate, biperiden, butropium bromide, buzepide, camylofine, caramiphen, chlorbenzoxamine, chlorphenoxamine, cimetropium bromide, clidinium bromide, cyclodrine, cyclonium, cyclopentolate, cycrimine, darifenacin, deptropine, dexetimide, dibutoline sulfate, dicyclomine, diethazine, difemerine, dihexyverine, diphemanil methylsulfate, dipiproverine, diponium, emepronium, emepronium bromide, endobenzyline, ethopropazine, ethybenztropine, ethylbenzhydramine, etomidoline, eucatropine, fenpiverinium bromide, fentonium, fentonium bromide, flavoxate, flutropium, flutropium bromide, glycopyrrolate, heteronium, hexocyclium methyl sulfate, homatropine, homatropine, methyl, bromide, hyocyamine, hyoscyamine, ipratropium, ipratropium bromide, isopropamide, isopropamide iodide, levomepate, mecloxamine, mepenzolate, mepenzolate bromide, metcaraphen, methantheline, methantheline bromide, methixene, methscopolamin bromide, n-(l,2-diphenylethyl)nicotinamide, n- butylscopolammonium bromide, octamylamine, oxitropium bromide, oxybutynin, oxyphencyclimine, oxyphenonium, oxyphenonium bromide, pentapiperide, penthienate, penthienate bromide, phencarbamide, phenglutarimide, pipenzolate, pipenzolate bromide, piperdolate, piperidolate, piperilate, poldine methylsulfate, pridinol, prifmium, procyclidine, profmium bromide, propantheline, propantheline bromide, propenzolate, propiverine, propyromazine, scopolamine, scopolamine n-oxide, stilonium, stramonium, sultroponium, telenzepine, thihexinol, thiphenamil, tiemonium, tiemonium iodide,

timepidium, timepidium bromide, tiotropiuin bromide, tiquizium, tiquizium bromide, tolterodine, tridihexethyl iodide, trihexyphenidyl hydrochloride, tropacine, tropenzile, tropicamide, trospium, trospium chloride, valethamate, valethamate bromide, xenytropium.

It is preferred for the corticosteroid to be selected from: 21-acetoxy- pregnenolone; alclometasone; algestone; amcinonide; beclomethasone; betamethasone; betamethasone valerate; budesonide; chloroprednisone; ciclesonide; clobetasol; clobetasol propionate; clobetasone; clobetasone butyrate; clocortolone; cloprednol; corticosterone; cortisone; cortivazol; deflazacort; desonide; desoximethasone; dexamethasone; diflorasone; diflucortolone; difluprednate; enoxolone; fluazacort; flucloronide; flumethasone; flumethasone pivalate; flunisolide; fluocinolone acetonide; fluorocinolone acetonide; fluorocortolone hexanoate; diflucortolone valerate; fluocinonide; fluocortine; butyl fluocortolone; fluorometholone; fluperolone acetate; fluprednidene acetate; fluprednisonole; flurandrenolide; fluticasone propionate; formocortal; halcinonide; halobetasol propionate; halometason; halopredone acetate; hydrocortamate; hydrocortisone; hydrocortisone acetate; hydrocortisone butyrate; hydrocortisone phosphate; hydrocortisone 21 -sodium succinate; hydrocortisone tebutate; loteprednol etabonate; mazipredone; medrysone; meprednisone; methylprednisolone; momethasone furoate; paramethasone; prednicarbate; prednisolone; prednisolone; 21- diethylaminoacetate; prednisolone sodium phosphate; prednisolone sodium succinate; prednisolone sodium 21-m-sulfobenzoate; prednisolone sodium 21-stearoylglycolate; prednisolone tebutate; prednisolone 21 -trimethylacetate; prednisone; prednival; prednylidene; prednylidene 21-diethylaminoacetate; rimexolone; tixocortol; triamcinolone; triamcinolone acetonoide; triamcinolone benetonide; triamcinolone hexacetonide

It is preferred for the calcium blocker to be selected from: (S)-emopamil; 8363-S; amiloride; amlodipine; amlodipine; anipamil; azidopine; benidipine; bepridil; caroverine; CD349; CERM-11956; cinnarizine; CV4093; D-600; D-888; DHP-218; diclofurime; dilfϊazine; diltiazem; dipropervine; emopamil; felodipine; fendiline; floridine; flunarizine; gallopamil; GX 1048; iodipine; isradipine; KW3049; lacidipine; lercanidipine; lidoflazine; MDL72567; mesudipine; mibefradil; mioflazine; nicardipine; nifedipine; niguldipine; niludipine; nilvadipine; nimodipine; nisoldipine; nitrendipine; nivaldipine; oxodipine; perhexiline; phenytoin; pimozide; isradipine; pranidipine; prenylamine; darodipine; R-56865; R-58735; ranolzine; Rol8-3981; ryosidine; Smith Kline 9512; TC81; terodiline; thioridazine;tiapamil; vatanidipine; verapamil;YM-09730-5; (4S)DHP. β ₂ -agonists give a fast but weak relaxation of small human bronchi. When these substances are given together with a compound of the invention that gives a strong but slowly developing relaxation, the result is a quickly developing, strong and long lasting relaxation. For instance, when combining the β ₂ -agonist terbutalin with a compound of the invention, the former is administered by inhalation in an amount of from 2 to 10 mg, preferably about 5 mg, up to 3 times per day.

Corticoteroids are one of the most important therapies in asthma. They reduce the inflammation in the airways, and reduce the bronchial hyperreactivity, thus reducing the need for additional bronchodilators. By the combined administration of steroid and a compund of the invention the inflammatory process is combatted and the tendency of the airways to contract spontaneously is reduced. For instance, the corticosteroid budesonide

can be administered in combination with a compound of the invention by inhalation in an amount of from 400-1600 μg/day.

Anticholinergic drugs are the preferred bronchodilators in patients with COPD (Chronic Obstructive Pulmonary Disease), although the relaxing effect is weak. If an anticholinergic is administered in combination with a compound of the invention the relaxing effect is markedly improved. The compounds of the invention have a pronounced relaxing effect on small human bronchi, which is the location for COPD-induced pathological changes. For instance, the anticholinergic ipratropium bromide is given in a dose of 40 μg 4 times per day in combination with a compound of the invention.

Antagonists of voltage operated calcium channels (VOC) have been tested as bronchodilators in asthma. While they give some relaxation of small human bronchi, this relaxation is much weaker than their relaxing effect on, for instance, small arteries. The bronchorelaxation by VOC antagonists on small human bronchi develops fairly quickly, but is gradually reduced in spite of a continuous presence of VOC inhibitors. However, if a VOC antagonist is administered to a patient in combination with a compound of the invention, the relaxation will be fast, strong and long lasting. For instance, the calcium channel blocker nifedipine is given in a dose of 40 mg 2 times per day in combination with a compound of the invention.

In general the anti-asthmatic selected from β ₂ -agonist, anticholinergic, corticosteroid, and calcium antagonist will be administered to a patient in combination with a compound of the invention in therapeutic amount corresponding to a dose from 0.1 to 1.0 of an established dose in which the β ₂ -agonist, anticholinergic, corticosteroid or calcium antagonist is therapeutically effective when administered alone.

According to the invention is also disclosed a pharmaceutical composition for the treatment of asthma and related conditions for oral administration selected from β ₂ - agonist, anticholinergic, corticosteroid, and calcium antagonist and a pharmaceutically acceptable carrier, the therapeutic amount of β ₂ -agonist, anticholinergic, corticosteroid or calcium antagonist in a single dose thereof corresponding to a dose from 0.1 to 1.0 of an established dose in which the β ₂ -agonist, anticholinergic, corticosteroid or calcium antagonist is therapeutically effective when administered alone.

Unless otherwise stated all parts in this specification are by weight.

DECRIPTION OF THE FIGURES

The invention is illustrated in a single Figure showing an exemplary test used for testing the compounds of the invention for their bronchoconstrictive properties.

DESCRIPTION OF PREFERRED EMBODIMENTS OF THE INVENTION

Synthesis of urea compounds of the invention OC=O^

EXAMPLE 1. Synthesis of N-[2-(4-chlorophenyl)ethyl]-6,7-dihydroxy-3,4- dihydroisoquinoline-2(17f)-carboxamide Res-11-79 and 5,8-dichloro-iV-[2-(4- chlorophenyl)ethyl]-6,7-dihydroxy-3,4-dihydroisoquinoline-2( lH)-carboxamide

Res-11-91.

The title compounds were synthesized according to Scheme 1.

Scheme 1. Synthesis ofN-[2-(4-chlorophenyl)ethyl]~6, 7-dihydroxy-3,4- dihydroisoquinoline-2(lH)-carboxamide Res-11-79 and 5,8-dichloro-N-[2-(4- chlorophenyl)ethyl]-6, 7-dihydroxy-3,4-dihydroisoquinoline-2(lH)-carboxamide Res-11-91.

Trichloroacetyl chloride (1.0 eq.), was dissolved in anhydrous THF under nitrogen, then 2-(4-chlorophenyl)ethyl amine (1) (1.0 eq.) was added dropwise to the solution. The reaction mixture was stirred at room temperature for 3.5 hours. The mixture was concentrated and the residue cliromatographed on silicagel (petroleum ether: EtOAc, 3:1) yielding 2,2,2-trichloro-N-[2-(2-chlorophenyl)ethyl]acetamide (Ia) as white crystals (53%).

Commercially available 6,7-dimethoxy-l ₅ 2,3,4-tetrahydroisoquinoline (2) (1.0 eq.) was suspended in glacial acetic acid and SO ₂ Cl ₂ (2.5 eq.) was added dropwise. After stirring for 2.5 hours the mixture was concentrated affording 5,8-dichloro-6,7-dimethoxy-l,2,3,4- tetrahydroisoquinoline (2a) as its hydrochloric salt quantitatively.

The corresponding amine (2 or 2a) was dissolved in HBr (48% in H ₂ O). The mixture was heated to 105°C for 5 hours and then concentrated. The residue was suspended in EtOAc and concentrated to afford the hydrobromic salt of either (2b) or (2c).

The hydrobromic salt of the corresponding bicyclic amine (2b or 2c) (1.0 eq.) was dissolved in anhydrous DMSO, DBU (1.0 eq.) was added and the solution stirred for 15 min. Then 2,2,2-trichloro-N-[2-(2-chloroρhenyl)ethyl]acetamide (Ia) (1.0 eq.) and DBU (1.0 eq.) were added. The intermediate 2-(4-chlorophenyl)ethyl isocyanate is formed in situ. The reaction mixture was stirred at 80° C for 48 hours. CH ₂ Cl ₂ was added to the solution and the organic phase was washed with HCl (3% in H ₂ O) and saturated solution OfNaHCO ₃ . The organic phase was dried (MgSO ₄ ) and concentrated. The residue was chromatographed on silicagel (2%MeOH in CH ₂ Cl ₂ ) yielding iV-[2-(4- chlorophenyl)ethyl]-6,7-dihydroxy-3,4-dihydroisoquinoline-2( lH)-carboxamide Res-ll- 79 (23%) or (gradient elution: 0.5-2%MeOH in CH ₂ Cl ₂ ) to yield 5,8-dichloro-λ ^r -[2-(4- chlorophenyl)ethyl]-6,7-dihydroxy-3,4-dihydroisoquinoline-2( lH)-carboxamide Res-ll- 91(38%).

EXAMPLE 2. Physical data of the compounds of the invention

General. ¹ H-NMR spectra and ¹³ C-NMR spectra were recorded with a Bruker DRX-400 (at 400/100 MHz). CD ₃ OD (3.31/49.0 ppm), was used as solvent for NMR (calibration value shown in parenthesis). ESI-MS spectra were recorded on a MicroMass Q-TOF Micro spectrometer.

Res-11.79. N-[2-(4-chlorophenyl)ethyl]-6, 7-dihydroxy-3,4-dihydroisoquinoline-2(lH)- carboxamide. ¹ H-NMR δ 2.64 (t, J=5.9 Hz, 2H), 2.77 (t. J=7.3 Hz, 2H), 3.37 (t, J=7.3 Hz, 2H), 3.51 (t, J=5.9 Hz, 2H) ₃ 4.33 (s, 2H), 6.53 (s, IH), 6.56 (s, IH), 7.15 (d, J=8.4 Hz, 2H), 7.22 (t, J=8.4 Hz, 2H). ¹³ C-NMR 5 29.0, 36.8, 42.7, 43.3, 46.2, 113.8, 116.0, 125.6, 127.1, 129.4, 129.4, 131.5, 131.5, 132.9, 139.7, 145.0, 145.2, 160.0. ESI-MS calculated for C ₁₈ H ₂₀ N ₂ O ₃ Cl (M+H) 347.1123, found 347.1162.

Res-11.91. 5, 8-dichloro-N-[2-(4-chlorophenyl)ethyl]-6, 7-dϊhydroxy-3, 4- dihydroisoquinoline-2(lH)-carboxamide. ¹ H-NMR δ 2.68 (t, J=5.8 Hz, 2H), 2.77 (t. J=7.3 Hz, 2H) ₅ 3.38 (t, J=7.3 Hz, 2H), 3.55 (t, J=5.8 Hz, 2H), 4.43 (s, 2H), 7.13 (d, J=8.3 Hz, 2H), 7.20 (t, J=8.3 Hz, 2H). ¹³ C-NMR δ 27.1, 36.7, 42.0, 43.2, 45.3, 118.5, 120.5, 124.6, 125.7, 129.4, 129.4, 131.5, 131.5, 133.0, 139.7, 142.5, 142.8, 159.8. ESI-MS calculated for C ₁₈ H ₁₈ N ₂ O ₃ Cl ₃ (M+H) 415.0301, found 415.0383. Synthesis of thiourea compounds of the invention (X-S)

EXAMPLE 3. Synthesis of N-[2-(4-chloroρhenyl)ethyl]-7-[(methylsulfonyl)amino]-3,4- dihydroisoquinoline-2(lH)-carbothioamide (Res-10-73) and N-[2-(4-chlorophenyl)ethyl]- 6- [(methylsulfonyl)amino] -3 ,4-dihydroisoquinoline-2(l H)-carbothioamide (Res-12-31).

The title compounds were synthesized according to Scheme 2.

H ₂ SCyHNO ₃

3a R ₁ =NO ₂ R ₂ =H

3b R;=H R ₂ =NO ₂

4a R ₁ =NH ₂ R ₂ =H 5a R ₁ =NH ₂ R ₂ =H 6a R ₁ =NH ₂ R ₂ =H

4b R ₁ =H R ₂ =NH ₂ 5b R ₁ =H R,=NH, 6b R ₁ =H R ₂ =NH ₂

Res-10-73 R ₁ =NSOXH, R ₀ =H

Res-12-31 R ₁ =H R ₂ =NSO ₂ CH ₃

Scheme 2. Synthesis ofN-[2-(4-chlorophenyl)ethyl]-7-[(methylsulfonyl)amino]-3,4- dihydroisoquinoline-2(lH)-carbothioamide (Res-10-73) and N-[2-(4-chlorophenyl)ethyl] - 6-[(methylsulfonyl)amino] -3 , 4-dihydroisoquinoline-2(lH)-carbothioamide (Res-12-31).

1,2,3,4-Tetrahydroisoquinoline (1) (1 eq.) was cooled on ice and acetic anhydride (1.5 eq.) was added dropwise. The mixture was stirred for 2 hours and then diluted with EtOAc. The organic phase was washed with NaHCO ₃ (sat.), dried (MgSO ₄ ) and concentrated to give 2-acetyl-l,2,3,4-tetrahydroisoquinoline (2) (58%).

2-Acetyl-l ₅ 2,3,4-tetrahydroisoquinoline (2) was cooled on ice and a 1:1 mixture of concentrated nitric and concentrated sulfuric acid was added dropwise. The mixture was stirred on ice for 4 hours and then poured into a mixture of ice and water. The water phase was extracted with EtOAc. The combined organic phases were washed with NaHCO ₃ (sat.), dried (MgSO ₄ ) and concentrated to give a crude mixture of regioisomers (84%). Pure isomers were obtained by HPLC (Microsorb, silica 5μm, 250x21.4 mm, 20ml/min of 100% EtOAc, detection at 300 nm): 2-acetyl-7-nitro-l,2,3,4-tetrahydroisoquinoline (3a) (21%) and 2-acetyl-6-nitro-l,2,3,4-tetrahydroisoquinoline (3b) (13%).

The corresponding 2-acetyl-mononitro-l,2,3,4-tetrahydroisoquinoline was dissolved in MeOH and some HCl (10% in water) and palladium on carbon (5%) was added. The mixture was stirred under hydrogen atmosphere for 1 hour, filtered through celite and concentrated to give the hydrochloride salts of 2-acetyl-l,2,3,4-tetrahydroisoquinolin-7- amine (4a) (89%) and 2-acetyl-l,2,3,4-tetrahydroisoquinolin-6-amine (4b) (quant.).

The hydrochloride salt of the corresponding 2-acetyl- 1,2,3 ,4-tetrahydroisoquinoline- monoamine was dissolved in HBr (48% in H ₂ O) and heated to reflux for 4 hours. The mixture was then concentrated to give the dihydrobromide salts of 1,2,3,4- tetrahydroisoquinolin-7-amine (5a) (91%) and l,2,3,4-tetrahydroisoquinolin-6-amine (5b) (80%). These salts were suspended in a 6M solution of NaOH and extracted with CH ₂ Cl ₂ . The organic phases were dried (MgSO ₄ ) and concentrated to give the free amines (quant.).

The corresponding 1,2,3,4-tetrahydroisoquinoline-monoamine (1 eq.) was dissolved in THF, di-tert-butyldicarbonate (1.2 eq.) was added. The solution was stirred at room temperature for 3 hours. The reaction mixture was concentrated, dissolved in H ₂ O and extracted with EtOAc. The organic phase was dried (MgSO ₄ ), filtered and concentrated. The residue was chromatographed on silicagel (Pet. Ether :EtO Ac 1:1) yielding tert-butyl 7-amino-3,4-dihydroisoquinoline-2(lH)-carboxylate (6a) (47%) and tert-butyl 6-amino- 3,4-dihydroisoquinoline-2(lH)-carboxylate (6b) (73%)

The corresponding tert-butyl amino-3,4-dihydroisoquinoline-2(l//)-carboxylate was dissolved in CH ₂ Cl ₂ and cooled on ice, MsCl (1.05 eq.) and triethylamine (1.15 eq.) were added. The solution was stirred for 2 hours, then diluted with H ₂ O and extracted with CH ₂ Cl ₂ . The organic phase was dried (MgSO ₄ ) and concentrated. The residue was chromatographed on silicagel (Pet. Ether: EtOAc 3:2) yielding tert-butyl 7- [(methylsulfonyl)amino]-3,4-dihydroisoquinoline-2(lH)-carbox ylate (7a) (43%) and tert- butyl 6-[(methylsulfonyl)amino]-3,4-dihydroisoquinoline-2(lϋ/)-ca rboxylate (7b) (75%).

The corresponding tert-butyl [(methylsulfonyl)amino]-3,4-dihydroisoquinoline-2(lH)- carboxylate was dissolved in a solution consisting in 80%TFA, 19%CHiCl ₂ and 1% anisol. This solution was stirred at 0°C for for 30 minutes and concentrated. The residue was dissolved in CH ₂ Cl ₂ and the mixture concentrated again. The residue was the dissolved in dry DMF and triethylamine (3 eq.) was added. This mixture was stirred for 30 minutes and then 2-(4-chlorophenyl)ethyl isothiocianate (1.1 eq.) was added. This mixture was stirred for 1 hour and then concentrated. The residue was dissolved in EtOAc and washed with 10% HCl solution. The organic phase was dried (MgSO ₄ ) and concentrated. The crude was chromatographed on silicagel (heptane:EtOAc:AcOH (40:60:1) yielding N- [2-(4-chlorophenyl)ethyl]-7-[(methylsulfonyl)amino]-3,4-dihy droisoquinoline-2(lH)- carbothioamide (Res-10-73) (66%) andN-[2-(4-chlorophenyl)ethyl]-6- [(methylsulfonyl)amino]-3,4-dihydroisoquinoline-2(lH)-carbot hioamide (Res-12-31) (56%).

EXAMPLE 4. Synthesis of tert-butyl 6,7-dicyano-3,4-dihydroisoquinoline-2(lH)- carboxylate and dimethyl l,2,3,4-tetrahydroisoquinoline-6,7-dicarboxylate

The title compounds were synthesized according to Scheme 3.

Scheme 3. Synthesis oftert-butyl 6, 7-dicyano-3,4-dihydroisoquinoline-2(lH)-carboxylate, dimethyl I ₁ 2, 3, 4-tetrahydroisoquinoline-6, 7-dicarboxylate

The commercially available 6,7-dimethoxy-l -methyl- 1, 2,3 ,4-tetrahydroisoquinoline (5.17g) hydrochloride (8) was refluxed in HBr (48% in H ₂ O) for 5 hours and concentrated to give 4.84 g (quant.) of l,2,3,4-tetrahydroisoquinoline-6,7-diol hydrobromide (9).

Without further purification 9 (3.77 g.) was suspended in H ₂ O (15 ml), di-tert- butyldicarbonate (3.51 mg, 1.1 eq.) and TEA (4.5 ml, 2.1 eq.) in 40 ml THF was added dropwise. The mixture was stirred overnight. The mixture was concentrated, dissolved in EtOAc and washed with water. The organic phase was dried (MgSO ₄ ), concentrated and chromatographed on silica (Petroleum Ether:EtOAc) to give 2.8 g. (68%) of tert-butyl 6,7- dihydroxy-3,4-dihydroisoquinoline-2(lH)-carboxylate (10). Trifluoromethanesulfonic anhydride (1.523 ml, 2.05 eq.) was added slowly to an ice cold solution of 10 (1.17g) and TEA (1.5 ml, 2.5 eq.) in dry CH ₂ Cl ₂ (25 ml). The mixture was allowed to reach rt. After 2 hours the mixture was poured into NaHCO ₃ (sat.). The water phase was extracted with CH ₂ Cl ₂ The organic phase was dried (MgSO ₄ ) and concentrated to give 2.2 g (95%) of tert-butyl 6,7-bis{[(trifluoromethyl)sulfonyl]oxy}-3,4- dihydroisoquinoline-2(l/i)-carboxylate (11).

To a mixture of 11 (2.2 g), tris(dibensylideneacetone)dipalladium (156 mg, 0.04 eq.) and 1,1 '-bis(diphenylphosphino)ferrocene (371 mg, 0.16 eq.) in DMF heated to 8O ⁰ C was Zn(CN) ₂ (30 mg, 0.06 eq.) added every 6:th minute over 2 hours (600 mg. 1.2 eq in total). The mixture was poured into Na ₂ CO ₃ (sat.) and the water phase extracted with CH ₂ Cl ₂ . The combined organic phases were washed with water, dried (MgSO ₄ ) and concentrated. The residue was chromatographed on silica (Petroleum Ether.εtOAc) to give 500 mg. (42%) oftert-butyl 6,7-dicyano-3,4-dihydroisoquinoline-2(lH)-carboxylate (12). A suspension of 12 (112 mg) in NaOH (50% in H ₂ O) was heated to reflux for 15 hours. The mixture was concentrated and the solid extracted with MeOH. To the organic phase was H ₂ SO ₄ added and it was refluxed for 15 hours. The mixture was concentrated. The residue was suspended in NaHCO ₃ . The water phase was extracted with EtOAc. The combined organic phases were dried (MgSO^ and concentrated to give 44 mg (35% over two steps) of dimethyl l,2,3,4-tetrahydroisoquinoline-6,7-dicarboxylate (13).

EXAMPLE 5. Synthesis of iV-[2-(4-chloroρhenyl)ethyl]-6,7-dicyano-3,4- dihydroisoquinoline-2( 1 H)-carbothioamide (Res-12-45) The title compound was synthesized according to Scheme 4.

Scheme 4. Synthesis ofN'[2-(4-chlorophenyl)ethyl]-6, 7-dicyano-3,4-dihydroisoquinoline- 2(lH)-carbothioamide (Res-12-45)

The di-nitrile 12 (51 mg) was dissolved in a mixture of TFA (80%), CH ₂ Cl ₂ (19%) and anisol (1%) and stirred for 20 minutes and then concentrated. The residue was was dissolved in DMF and triethylamine (75 μl, 3 eq.) was added. This mixture was stirred for 15 minutes and then was 2-(4-chlorophenyl)ethyl isothiocianate (30 μl, 1.2 eq.) added. This mixture was stirred for additional 3 hours and then concentrated. The residue was dissolved in EtOAc and washed with water. The organic phase was dried (MgSO ₄ ), concentrated and chromatographed on silicagel (heptane:EtOAc) to give 43 mg (63% over two steps) of N-[2-(4-chlorophenyl)ethyl]-6,7-dicyano-3,4-dihydroisoquinol ine-2(lH)- carbothioamide (Res-12-45).

EXAMPLE 6. Synthesis of dimethyl 2-({[2-(4-chlorophenyl)ethyl]ammo}carbonothioyl)- 1 ,2,3,4-tetrahydroisoquinoline-6,7-dicarboxylate (Res-12-69)

The title compound was synthesized according to Scheme 5.

Scheme 5. Synthesis of dimethyl 2-({[2-(4-chlorophenyl)ethyl]amino}carbonothioyl)- 1, 2, 3, 4-tetrahydroisoquinoline-6, 7-dicarboxylate (Res-12-69)

The di-ester 13 (25 mg) was dissolved in DMF and triethylamine (42 μl, 3 eq.) was added. This mixture was stirred for 15 minutes and then was 2-(4-chlorophenyl)ethyl isothiocianate (18 μl, 1.2 eq.) added. This mixture was stirred for additional 3 hours and then concentrated. The residue was dissolved in EtOAc and washed with water. The organic phase was dried (MgSO ₄ ), concentrated and chromatographed on silicagel (heptane:EtOAc) to give Res-12-69 (17 mg, 38%).

EXAMPLE 7. Synthesis of 2-({[2-(4-chlorophenyl)ethyl]amino}carbonothioyl)-l,2,3,4- tetrahydroisoquinoline-6,7-dicarboxylic acid (Res- 12-85)

The title compound was synthesized according to Scheme 6.

Res-12-85

Scheme 6. Synthesis of ^' 2-({[2~(4-chlorophenyl) ethyl] ' amino}carbonothioyl)-l ,2,3 ,4- tetrahydroisoquinoline-6, 7-dicarboxyIic acid (Res-12-85).

To a solution of the di-ester 13 (39 mg) in 10 ml THF/ H ₂ O was LiOH*H ₂ O (33 mg, 5 eq.) added. The mixture was stirred at rt for 3 days then was HCl (10% in H ₂ O) added. The resulting acidic mixture was concentrated to give l,2,3,4-tetrahydroisoquinoline-6,7- dicarboxylic acid hydrochloride (14) which was coupled to 2-(4-chlorophenyl)ethyl isothiocianate without further purification.

The di-carboxylic acid 14 (40 mg) was dissolved in DMF and triethylamine (110 μl, 5 eq.) was added. This mixture was stirred for 15 minutes and then was 2-(4-chlorophenyl)ethyl isothiocianate (29 μl, 1.2 eq.) added. This mixture was stirred for additional 3 hours and then concentrated. The residue was pardoned between EtOAc and HCl (1% in H ₂ O). The water phase was extracted with EtOAc. The combined organic phases were dried (MgSO ₄ ), concentrated and washed with diethylether to give 18 mg, (28% over two steps) of 2-({ [2-(4-chlorophenyl)ethyl] amino }carbonothioyl)-l ,2,3,4~tetrahydroisoquinoline-6,7- dicarboxylic acid (Res-12-85).

EXAMPLE 8. Physical data of the compounds of the invention

General. ^l H-NMR spectra and ¹³ C-NMR spectra were recorded with either of the following spectrometers: Bruker 300-DRX (at 300/75 MHz), Bruker DRX-400 (at 400/100 MHz) or Bruker ARX-500 (500/125 MHz). CD ₃ OD (3.31/49.0 ppm), CDCl ₃ (7.26/77.2 ppm) and (CD ₃ ) ₂ SO (2.50/39.5 ppm) were used as solvents for NMR (calibration value shown in parenthesis). ESI-MS spectra were recorded on a MicroMass Q-TOF Micro spectrometer. Otherwise mentioned, all compounds were obtained as oils.

Res-10-73. N-[2-(4-chlorophenyl)ethyl]-7-[(methylsulfonyl)amino]-3,4- dihydroisoquinoline-2(lH)-carbothioamide. ¹ H-NMR (CDCl ₃ 300 MHz) δ 2.86 (t, J=5.6 Hz, 2H), 2.97 (m, 5H), 3.87 (t, J=5.8 Hz, 2H), 3.94 (m, 2H) 4.85 (s, 2H), 7.09 (m, 3H), 7.18 (d, J=8.3 Hz, IH), 7.27 (d, J=8.3 Hz, IH), 7.38 (bs, IH). ¹³ C-NMR (CDCl ₃ 75 MHz) δ 28.3, 34.8, 39.4, 45.4, 47.0, 49.3, 119.2, 120.2, 128.8, 129.2, 130.3, 132.3, 132.5, 134.5, 135.4, 137,5, 181.2. ESI-MS calculated for C ₁₉ H ₂₃ N ₃ O ₂ S ₂ Cl (M+H) 424.0920, found 424.0918.

Res-12-31. N-[2-(4-chlorophenyl)ethyl]-6-[(methylsulfonyl)amino]-3,4- dihydroisoquinoline-2(lH)-carbothioamide. ¹ H-NMR (CDCl ₃ 400 MHz) δ 2.89 (t, J=5.9 Hz, 2H), 2.96 (t, J=6.9 Hz, 2H), 3.00 (s, 3H), 3.82 (t, J=5.9 Hz, 2H), 3.95 (dt, J=6.9, J=5.5 Hz, 2H), 4.85 (s, 2H), 5.54 (bs, IH), 6.89 (s, IH), 7.09 (m, 3H), 7.16 (d, J=8.3 Hz, IH), 7.27 (d, J=8.3 Hz, IH). ¹³ C-NMR (CDCl ₃ 100 MHz) δ 29.0, 34.8, 39.6, 45.1, 46.9, 49.1, 119.5, 120.3, 127.9, 128.9, 128.9, 130.3, 130.3, 130.5, 132.6, 135.9, 137,0,137.5, 181.5. ESI-MS calculated for C ₁₉ H ₂₃ N ₃ O ₂ S ₂ Cl (M+H) 424.0920, found 424.0929.

Res-12-45. N-[2-(4-chlorophenyl)ethyl]-6, 7-dicyano-3, 4-dihydroisoquinoline-2(lH)- carbothioamide. ¹ H-NMR (CDCl ₃ 300MHz) δ 2.95 (t, J=7.0 Hz, 2H), 3.03 (t, J=5.9 Hz, 2H), 3.85 (t, J=5.9 Hz, 2H) ₅ 3.93 (m, 2H), 5.07 (s, 2H), 5.68 (bs, IH), 7.15 (d, J=8.4 Hz, 2H), 7.26 (d, J=8.4 Hz, 2H), 7.61 (s, IH), 7.64 (s, IH). ¹³ C-NMR (CD ₃ OD 75 MHz) δ 28.9, 34.5, 44.0, 47.0, 49.3, 114.0, 114.3, 115.3, 115.3, 129.0, 129.0, 130.3, 130.3, 131.8, 132.6, 133.4, 137.3, 140.0, 141.7, 182.1. ESI-MS calculated for C ₂₀ H ₁₈ ClN ₄ S (M+H) 381.0941, found 381.0943.

Res-12-69. dimethyl 2-({[2-(4-chlorophenyl)ethyl]amino}carbonothioyl)-l, 2, 3, 4- tetrahydroisoquinoline-6, 7-dicarboxylate. ¹ H-NMR (CDCl ₃ , 400MHz) δ 2.95 (m, 4H), 3.88 (m, 2H), 3.89 (s, 3H) ₅ 3.90 (s, 3H), 3.94 (m, 2H), 4.91 (s, 2H), 5.59 (bs, IH), 7.15 (d, J=8.4 Hz, 2H), 7.26 (d, J=8.4 Hz, 2H), 7.52 (s, IH), 7.53 (s, IH). ¹³ C-NMR (CD ₃ OD 100 MHz) δ 28.8, 34.7, 44.9, 46.9, 49.1, 53.8, 53.8, 127.4, 128.8, 129.0, 129.0, 130.3, 130.3, 131.1, 131.1, 132.6, 136.4, 137.4, 138.8, 167.6, 167.9, 181.9. ESI-MS calculated for C ₂₂ H ₂₃ ClN ₂ NaO ₄ S (M+Na) 469.0965, found 469.0974. Res- 12-85. 2-({[2-(4-chlorophenyl)ethyl]amino}carbonothioyl)-l, 2, 3, 4- tetrahydroisoquinoline-6, 7-dicarboxylic acid. ¹ H-NMR (CD ₃ OD 300MHz) δ 2.96 (m, 4H), 3.84 (t, J=7.4 Hz, 2H), 4.03 (t, J=5.7 Hz, 2H), 4.96 (s, 2H), 7.21 (d, J=8.4 Hz, 2H), 7.25 (d, J=8.4 Hz, 2H), 7.60 (s, IH), 7.62 (s, IH). ¹³ C-NMR (CD ₃ OD 75 MHz) δ 29.4, 35.6, 46.3, 48.0, 50.3, 128.5, 129.5, 129.5, 130.3, 131.5, 131.5, 132.1, 132.9, 133.0, 137.8, 139.6, 140.0, 170.0, 171.3, 182.6. ESI-MS calculated for C ₂₀ H ₂₀ ClN ₂ O ₄ S (M+H) 419.0832, found 419.0819.

EXAMPLE 9. Synthesis of 5,8-dichloro-N-[2-(4-chloroρhenyl)ethyl]-6-hydroxy-7- methoxy-3,4-dihydroisoquinoline-2(lH)-carbothioamide (Res-13-61) and 5,8-dichloro-λ/- [2-(4-chlorophenyl)ethyl]-7-hydroxy-6-methoxy-3,4-dihydroiso quinoline-2(lH)- carbothioamide (Res-14-8)

The title compounds were synthesized according to Scheme 7.

Scheme 7. Synthesis of 5 ,8-dichloro-N-[2-(4-chlorophenyl)ethyl]-6-hydroxy-7 -rneihoxy- 3,4-dihydroisoquinoline-2(lH)-carbothioamide (res-13-61) and 5,8-dichloro-N-[2-(4- chlorophenyl)ethyl]-7-hydroxy-6-methoxy-3,4-dihydroisoquinol ine-2(lH)-carbothioamide (Res-14-8)

7-methoxy-l,2,3,4-tetrahydroisoquinolin-6-ol hydrochloride (1) and 6-methoxy-l,2,3,4- tetrahydroisoquinolin-7-ol hydrochloride (2) were synthesized from vanillin and isovanillin respectively according to a procedure described by Bobbitt et al (J. Org. Chem. (1965) 30 2247-2250)

To a solution of the corresponding amine (1 or 2) (leq.) in acetic acid (glacial) was SO ₂ Cl ₂ (2.5 eq.) added dropwise. After stirring for 5 hours the mixture was concentrated to give 5,8-dichloro-7-methoxy-l,2,3,4-tetrahydroisoquinolin-6-ol hydrochloride (Ia) and 5,8-dichloro-6-methoxy-l,2,3,4-tetrahydroisoquinolin-7-ol hydrochloride (2a) which were coupled to 2-(4-chlorophenyl)ethyl isothiocianate without further purification.

The corresponding tetrahydroisoquinoline (Ia or 2a) (1 eq.) was dissolved in anhydrous DMF and triethylamine (3 eq.) was added. This mixture was stirred for 15 minutes and then was 2-(4-chlorophenyl)ethyl isothiocianate (1.2 eq.) added. This mixture was stirred for additional 3 hours and then concentrated. The residue was dissolved in EtOAc and washed with water. The organic phase was dried (MgSO ₄ ), concentrated and chromatographed on silicagel (Petroleum Ether:EtOAc+AcOH, 4:1+1%) to give 5,8- dichloro-iV-[2-(4-chlorophenyl)ethyl]-6-hydroxy-7-methoxy-3, 4-dihydroisoquinoline- 2(l#)-carbothioamide, Res-13-61 (40 %) or chromatographed on silicagel (Petroleum Ether:EtOAc+AcOH, 4:2+1%) to give 5,8-dichloro-iV-[2-(4-chlorophenyl)ethyl]-7- hydroxy-6-methoxy-3,4-dihydroisoquinoline-2(17f)-carbothioam ide, Res-14-8 (28%)

EXAMPLE 10. Physical data of the compounds of the invention General. ¹ H-NMR spectra and ¹³ C-NMR spectra were recorded with the following spectrometer, Broker DRX-400 (at 400/100 MHz). CD ₃ OD (3.31/49.0 ppm) and CDCl ₃ (7.26/77.2 ppm) were used as solvents for NMR (calibration value shown in parenthesis). ESI-MS spectra were recorded on a MicroMass Q-TOF Micro spectrometer. Res-13-61. 5,8-dichloro-N-[2-(4-chlorophenyl)ethyl]-6-hydroxy-7-methoxy -3,4- dihydroisoquinoline-2(lH)-carbothioamide. ¹ H-NMR (CDCl ₃ 400 MHz) δ 2.91 (t, J=5.9 Hz, 2H), 2.99 (t, J=7.0 Hz, 2H), 3.94 (s, 3H), 3.97 (m, 2H), 4.04 (t, J-5.9 Hz, 2H), 4.72 (s, 2H), 5.75 (t, J=5.0 Hz, IH), 7.20 (d, J=8.4 Hz, 2H), 7.30 (d, J=8.4 Hz, 2H). ¹³ C-NMR (CDCl ₃ 100 MHz) 5 26.5, 34.7, 44.9, 47.0, 47.2, 61.3, 119.0, 123.3, 124.1, 129.0, 129.1, 129.1, 130.3, 130.3, 132.6, 137.5, 142.8, 145.4, 181.9. ESI-MS calculated for C ₁₉ H ₂₀ Cl ₃ N ₂ O ₂ S 445.0311 (M+H), found 445.0304.

Res-14-8. 5, 8-dichloro-N-[2-(4-chlorophenyl)ethyl]-7~hydroxy-6-methoxy-3 , 4- dihydroisoquinoline-2(lH)-carbothioamide. ¹ H-NMR (CD ₃ OD 400 MHz) δ 2.78 (t, J=5.9 Hz, 2H), 2.93 (t, J=7.4 Hz, 2H), 3.81 (m, 5H), 3.95 (t, J=5.9 Hz, 2H), 4.89 (s, 2H), 7.17 (d, J=8.7 Hz, 2H), 7.21 (d, J=8.7 Hz, 2H). ¹³ C-NMR (CD ₃ OD 100 MHz) δ 27.0, 35.5, 45.7, 47.9, 49.8, 61.2, 119.1, 126.1, 127.2, 129.4, 129.4, 129.4, 131.5, 131.5, 133.0, 139.5, 145.0, 147.4, 182.7. ESI-MS calculated for C ₁₉ H ₂₀ Cl ₃ N ₂ O ₂ S 445.0311 (M+H), found 445.0301.

EXAMPLE 11. Synthesis of 5,8-dichloro-6,7-dihydroxy-N-[2-(l-oxidoρyridin-2-yl)ethyl] - 3 ,4-dihydroisoquinoline-2( 1 H)-carbothioamide (Res-13-55).

The title compound was synthesized according to Scheme 8.

NCS

HBr (48%H ₂ O)

Scheme 8. Synthesis ofRes-13-55. 5,8-dichloro-6, 7-dihydroxy-N-[2-(l-oxidopyridin-2- yl)ethyl]-3,4-dihydroisoquinoline-2(lH)-carbothioamide. (Res-13-55)

2-Pyridin-2-ylethylamine (1) was dissolved in anhydrous DMF. To this solution was di- tert-butyldicarbonate (1.2 eq.) added. The solution was stirred at room temperature for 1 hour. The reaction mixture was diluted with H ₂ O and extracted with CH ₂ Cl ₂ . The organic phase was washed with H ₂ O, dried (MgSO ₄ ) and concentrated. The residue was dissolved in CHCl ₃ , the solution was cooled to 0 ⁰ C and rø-chloroperbenzoic acid (mCPBA) (1.0 eq.) dissolved in CHCl ₃ was added. The reaction mixture was stirred for 3.5 hours. During this time it reached room temperature. The reaction mixture was diluted with NaHCO ₃ (sat) and extracted with EtOAc. The organic phase was washed with brine, dried (MgSO ₄ ) and concentrated to give tert-butyl 2-(l-oxidopyridin-2-yl)ethylcarbamate (Ia) (quantitative).

(Ia) was dissolved in a solution consisting of 80%TFA, 19%CH ₂ C1 ₂ and 1% anisol. This solution was stirred at room temperature for 1 hour and concentrated. The residue was dissolved in anhydrous DMF and added to a solution of 1,1 '-thiocarbonyldiimidazole (1.2 eq.) in DMF at 4O ⁰ C. Finally triethylamine (2.0 eq.) was added. The reaction mixture was stirred at 4O ⁰ C for 3 hours, then diluted with H ₂ O and extracted with EtOAc. The combined organic phases were washed with H ₂ O, dried with MgSO ₄ and concentrated affording 2-(2-isothiocyanatoethyl)pyridine-l -oxide (Ib) which was used without further purification.

Commercially available 6,7-dimethoxy-l, 2,3, 4-tetrahydroisoquino line (2) (1.0 eq.)was suspended in glacial acetic acid and SO ₂ Cl ₂ (2.5 eq.) was added dropwise. After stirring for 2.5 hours the mixture was concentrated affording 5,8-dichloro-6,7-dimethoxy-l,2,3,4- tetrahydroisoquinoline (2a) as its hydrochloric salt, which was used without further purification.

(2a) was dissolved in HBr (48% in H ₂ O). The mixture was heated to 105°C for 5 hours and then concentrated. The residue was suspended in EtOAc and concentrated to afford the corresponding hydrobromic salt of 5,8-dichloro-l,2,3,4-tetrahydroisoquinoline-6,7- diol (2b) (quantitative).

(2b) (1.0 eq.) was dissolved in anhydrous DMF and triethylamine (3.0 eq.) was added. This mixture was stirred for 15 minutes and then 2-(2-isothiocyanatoethyl)pyridme 1- oxide (Ib) (1.2 eq.) was added. This mixture was stirred for additional 5 hours and then concentrated. The residue was dissolved in EtQAc and washed with water. The organic phase was dried (MgSO ₄ ) and concentrated. The crude was chromatographed on silicagel (heptane:EtOAc:AcOH (80:20:1) affording 5,8-dichloro-6,7-dihydroxy-iV-[2-(l- oxidopyridin-2-yl)ethyl] -3 ,4-dihydroisoquinoline-2( 1 H)-carbothioamide (Res-13-55) (54%), as a yellow solid. EXAMPLE 12. Physical data of the compound of the invention

General. ¹ H-NMR spectrum and ¹³ C-NMR spectrum were recorded with a Bruker DRX- 400 (at 400/100 MHz) spectrometer, (CD ₃ ) ₂ SO (2.50/39.5 ppm) was used as solvent for NMR (calibration value shown in parenthesis). ESI-MS spectrum was recorded on a MicroMass Q-TOF Micro spectrometer.

Res-13-55. 5, 8-dichloro-ό, 7-dihydroxy-N-[2-(l-oxidopyridin-2-yl)ethyl]-3, 4- dihydroisoquinoline-2(lH)-carbothioamide. ¹ H-NMR δ 2.67 (t, J=5.8 Hz, 2H), 3.18 (t, J=7.0 Hz ₅ 2H), 3.80 (m, 2H), 3.89 (t, J=5.8 Hz, 2H), 4.89 (s, 2H), 7.33 (m, 2H), 7.68 (dd, J- 7.4, 2.3 Hz, IH), 8.31 (d, J=5.9 Hz, IH), 8.48 (t, J=4.3 Hz, IH). ¹³ C-NMR δ 25.2, 29.0, 43.3, 43.5, 47.6, 117.0, 118.9, 123.0, 123.8, 124.3, 125.5, 126.4, 138.7, 141.0, 141.3, 149.0, 180.5. ESI-MS calculated for C ₁₇ H ₁₈ Cl ₂ N ₃ O ₃ S (M+H) 414.0449, found 414.0446

EXAMPLE 13. Synthesis of N-[2-(4-chlorophenyl)ethyl]-5-hydroxy-l,3-dihydro-2H- isoindole-2-carbothioamide (Res 12-83)

The title compound was synthesized according to Scheme 9.

N-bromosuccinimide Benzoyl peroxide

I) HBr (48% In H ₂ O), phenol, propionic acid

2) HBr (48% in H ₂ O)

Res-12-83

Scheme 9. Synthesis ofN-[2-(4-chlorophenyl)ethyl]-5~hydroxy-l,3-dihydro-2H-isoin dole- 2-carbothioamide (Res-12-83)

Tosylamide Monosodium Salt (TsNHNa). To a stirred refluxing solution of freshly prepared NaOEt (1.0 eq.) in absolute EtOH was added tosylamide (1.0 eq.). The mixture was refluxed for 2 hours and then cooled. The insoluble TsNHNa was collected by filtration, washed with absolute ethanol and dried in vacuo.

A mixture of 3,4-dimethylanisole (1), (1.0 eq.), N-bromosuccinimide (2.0 eq.) and benzoyl peroxide (cat.) was refluxed in CCl ₄ for 20 hours. After cooling, the insoluble material was filtered off and extracted with a small amount of CCl ₄ . The filtrate and CCl ₄ used for the extraction were mixed and concentrated to give an oily residue containing 3,4-bis-(bromomethyl)anisole (2).

To a stirred solution of TsNHNa (1.0 eq.) in anhydrous DMF at 8O ⁰ C was added dropwise under a N ₂ atmosphere .a solution of 2 (1.0 eq.) in DMF. The reaction mixture was stirred for 1 h. Then solid TsNHNa (1.0 eq.) was added at once and the mixture was stirred at 8O ⁰ C for 4 h. The reaction mixture was then concentrated and the solid residue was extracted with CHCl ₃ . The organic phase was washed with IM NaOH, dried (MgSO ₄ ) and concentrated. The solid residue was washed with MeOH and dried under reduced pressure yielding 5-methoxy-2-tolylsulfonylisoindoline (3). Compound 3 (1.0 eq.), phenol (2.5 eq.) and propionic acid (0.5 eq.) were dissolved in HBr (48% in H ₂ O), the mixture was refluxed for 4 hours under vigorous stirring under N ₂ . The reaction solution was concentrated and HBr (48% in H ₂ O) was added to the residue. The mixture was again refluxed under N ₂ for 3 hours. The reaction solution was cooled, and H ₂ O and CHCl ₃ added. The water phase was separated and treated with active carbon. The water phase was then concentrated and the crystalline residue washed with diethylether to afford the hydrobromic salt of 5-hydroxyisoindoline (4).

The amine 4 (1.0 eq.) was dissolved in anhydrous DMF and triethylamine (3.0 eq.) was added. This mixture was stirred for 15 minutes and then 2-(4-chlorophenyl)ethyl isothiocianate (1.2 eq.) was added. The mixture was stirred for additional 5 hours and then concentrated. The residue was dissolved in EtOAc and washed with water. The organic phase was dried (MgSO ₄ ) and concentrated to give the crude product, which was chromatographed on silicagel (heptane:EtOAc 8:2+l%AcOH) to afford Res-12-83 (31%). EXAMPLE 14. Physical data of the compound of the invention

General. ¹ H-NMR spectrum and ¹³ C-NMR spectrum were recorded with a Bruker DRX- 400 (at 400/100 MHz). CD ₃ OD (3.31/49.0 ppm), was used as solvent for NMR (calibration value shown in parenthesis). ESI-MS spectrum was recorded on a MicroMass Q-TOF Micro spectrometer.

Res-12-83. N-[2-(4-chlorophenyl)ethyl]-5-hydroxy-l,3-dihydro-2H-isoindo le-2- carbothioamide. ¹ H-NMR (CD ₃ OD 400 MHz) δ 2.94 (t, J=7.5 Hz ₃ 2H), 3.80 (t, J=7.5 Hz, 2H), 4.63 (bs, 4H), 6.73 (m, 2H), 7.10 (d, J-8.2 Hz, IH), 7.25 (m, 4H). ¹³ C-NMR (CD ₃ OD 100 MHz) δ 35.9, 47.7, 54.5, 57.9, 110.1, 116.2, 124.4, 127.6, 129.5, 129.5, 131.5, 131.5, 133.0, 138.5, 139.6, 158.6, 179.9. ESI-MS calculated for C ₁₇ H ₁₈ ClN ₂ OS (M+H) 333.0828, found 333.0837.

EXAMPLE 15. Synthesis of N-[2-(4-chlorophenyl)ethyl]-6,8-dihydroxy-3,4- dihydroisoquinoline-2(lH)-carbothioamide (Res-14-84)

The title compound was synthesized according to Scheme 10.

CH ₂ CI ₂ OMe

BBr ₃₁ CH ₂ CI ₂

Res-14-84

Scheme 10. Synthesis ofN-[2-(4-chlorophenyl)ethyl]-6,8-dihydroxy-3,4- dihydroisoquinoline-2(lH)-carbothioamide (Res-14-84)

2-(3,5-Dimethoxyphenyl)ethylamine, 1, (1.0 eq.), and MgSO ₄ (3.0 eq.) were suspended in anhydrous CH ₂ Cl ₂ , paraformaldehyde (5.0 eq.) was then added to the suspension in portions. After stirring for 2 hours the solid was filtered off. The filtrate was concentrated. The residue was dissolved in CHCl ₃ (2.5 ml/mmol of 1). This solution was slowly added to a cooled mixture of trifluoroacetic acid and CHCl ₃ (5:8) (20 ml/mmol of 1) under N ₂ . Once addition was finished, the mixture was refluxed under nitrogen for 5h. The mixture was poured into a mixture of ice and water. The water phase was made basic with NaOH (6M) and extracted with CH ₂ Cl ₂ . The organic phase was dried (MgSO ₄ ) and concentrated. The remaining oil was dissolved in anhydrous DMF. To this solution were added άi-tert- butyl dicarbonate (1.2 eq.) and triethylamine (3.0 eq.). The mixture was stirred for 3 hours and then concentrated. The residue was dissolved in EtOAc and washed with a saturated solution OfNa ₂ CO ₃ . The organic phase was dried (MgSO ₄ ) and concentrated. Purification was done by flash column chromatography (silica, Petroleum ether, EtOAc (9:1)) affording 2 (46%).

Compound 2 was dissolved in CH ₂ Cl ₂ (3 ml/mmol of 2) and cooled to -78 ⁰ C, then BBr ₃ (IM in CH ₂ Cl ₂ ) (10 eq.) was added dropwise. The mixture was then stirred overnight under N ₂ at r.t. H ₂ O was added to the mixture and then neutralized with aq. NaOH (IN), extracted with CH ₂ Cl ₂ , dried (MgSO ₄ ) and concentrated yielding 3 (42%).

The amine 3 (1 eq.) was dissolved in anhydrous DMF and triethylamine (3 eq.) was added. This mixture was stirred for 15 minutes and then 2-(4-chlorophenyl)ethyl isothiocianate (1.2 eq.) was added. This mixture was stirred for 6 hours and then concentrated. The residue was dissolved in EtOAc and washed with water. The organic phase was dried (MgSO ₄ ), concentrated and purified by flash column chromatography (silica, Petroleum ether:EtOAc (3:2)) to give iV-[2-(4-chlorophenyl)ethyl]-6,8-dihydroxy- 3,4-dihydroisoquinoline-2(lH)-carbothioamide (Res-14-84) (79%).

EXAMPLE 16. Physical data of the compound of the invention

General. ¹ H-NMR spectrum and ¹³ C-NMR spectrum were recorded with the following spectrometer: Bruker DRX-400 (at 400/100 MHz). CD ₃ OD (3.31/49.0 ppm) was used as solvent for NMR (calibration value shown in parenthesis). ESI-MS spectrum was recorded on a MicroMass Q-TOF Micro spectrometer.

Res-14-84. N-[2-(4-chlorophenyl)ethyl]-6, 8-dihydroxy-3, 4-dihydroisoquinoline-2(lH)- carbothioamide. ¹ H-NMR (CD ₃ OD 400MHz) δ 2.73 (t, J=5.8 Hz ₅ 2H), 2.92 (t, J=7.2 Hz ₅ 2H) ₅ 3.82 (t, J=7.2 Hz ₅ 2H) ₅ 3.99 (t ₅ J=5.8 Hz, 2H) ₅ 4.61 (s, 2H) ₅ 6.14 (d ₅ J=2.2 Hz ₅ 2H) ₅ 6.19 (d, J=2.2 Hz, 2H) ₅ 7.20 (d, J=8.7 Hz ₅ 2H) ₅ 7.24 (d, J=8.7 Hz, 2H). ¹³ C-NMR (CD ₃ OD 100 MHz) δ 30.0, 35.8, 45.7, 47.1, 47.9, 101.2, 106.8, 112.3, 129.4, 129.4, 131.5, 131.5, 133.0, 138.3, 139.7, 155.8, 157.8, 182.0. ESI-MS calculated for C ₁₈ H ₂₀ ClN ₂ O ₂ S (M+H) 363.0934, found 363.0901

EXAMPLE 17. Synthesis of N-[2-(4-chlorophenyl)ethyl]-6-hydroxy-7-nitro-3,4- dihydroisoqumoline-2(lH)-carbothioamide (Res-14-54) and iV-[2-(4-chlorophenyl)ethyl]- 6-hydroxy-5-nitro-3,4-dihydroisoqumoline-2(lH)-carbothioamid e (Res-14-56)

The title compounds were synthesized according to Scheme 11.

1 ) paraformaldehyde, MgSOφ CH2CI2 Boc

2) TFA Acetyl nitrate,

3) Boc anhydride, TEA, DMF Nitromethane

3a R ₁ = NO ₂ R ₂ = H 4a R ₁ = NO ₂ R ₂ = H 3b R ₁ = H R ₂ = NO ₂ 4b R ₁ = H R ₂ = NO ₂

H

Scheme 11. Synthesis ofN-[2-(4-chlorophenyl)ethyl]-6-hydroxy-7-nitro-3,4- dihydroisoquinoline-2(lH)-carbothioamide (Res-14-54) and N-[2-(4-chlorophenyl)ethyl]- 6-hydroxy-5-nitro-3,4-dihydroisoquinoline-2(lH)-carbothioami de (Res-14-56)

2-(3-Methoxyphenyl)ethylamine, 1, (1.0 eq.), and MgSO ₄ (3.0 eq.) were suspended in anhydrous CH ₂ Cl ₂ , paraformaldehyde (5.0 eq.) was then added to the suspension in portions. After stirring for 2 hours the solid was filtered off. The filtrate was concentrated. The residue was dissolved in anhydrous trifluoroacetic acid and refluxed under nitrogen over night. The mixture was poured into a mixture of ice and water. The water phase was made basic with NaOH (6M) and extracted with CH ₂ Cl ₂ . The organic phase was dried (MgSO ₄ ) and concentrated. The remaining oil was dissolved in anhydrous DMF. To this solution were added di-tert-butyl dicarbonate (1.2 eq.) and triethylamine (3.0 eq.). The mixture was stirred for 3 hours and then concentrated. The residue was dissolved in

EtOAc and washed with a saturated solution OfNa ₂ CO ₃ . The organic phase was dried (MgSO ₄ ) and concentrated. Purification was done by flash column chromatography (silica, gradient elution, 10-30% EtOAc in Petroleum Ether) affording 2 (47%).

Concentrated HNO ₃ (1.5 eq.) was added to acetic anhydride (6 eq.) under N ₂ at -2O ⁰ C. This solution was added to a solution of 2 in nitromethane under N ₂ at -20 ⁰ C. The reaction mixture was stirred at this temperature under N ₂ atmosphere for 3 hours. Toluene was added and the solution was washed with saturated solution OfNaHCO ₃ and brine. Then it was dried (MgSO ₄ ) and concentrated. Purification was done by flash column chromatography (silica, gradient elution 20-50% EtOAc in Petroleum ether) affording 3a and 3b in a 1:1 mixture (66%)

The corresponding amine (3a or 3b) were dissolved in HBr (48% in H ₂ O) and refluxed for 12 hours, Then it was concentrated to dryness to afford the hydrobromic salts of 4a or 4b quantitatively.

The amine 4a (or 4b) (1 eq.) was dissolved in anhydrous DMF and triethylamine (3 eq.) was added. This mixture was stirred for 15 minutes and then 2-(4-chlorophenyl)ethyl isothiocianate (1.2 eq.) was added. This mixture was stirred for 5 hours and then concentrated. The residue was dissolved in EtOAc and washed with water. The organic phase was dried (MgSO ₄ ), concentrated and purified by flash column chromatography (silica, Petroleum ether:EtOAc (3:2)) to give JV-[2-(4-chlorophenyl)ethyl]-6-hydroxy-7- nitro-3,4-dihydroisoquinoline-2(lH)-carbothioamide (Res-14-54) (76%) or (silica, CH ₂ Cl ₂ , Petroleum ether (8 :2)) to give JV-[2-(4-chlorophenyl)ethyl]-6-hydroxy-5-nitro- 3,4-dihydroisoquinoline-2(l/i)-carbothioamide (Res-14-56) (40%)

EXAMPLE 18. Physical data of the compounds of the invention General. H-NMR spectra and C-NMR spectra were recorded with the following spectrometer, Broker DRX-400 (at 400/100 MHz). CDCl ₃ (7.26/77.2 ppm) was used as solvent (calibration value shown in parenthesis). ESI-MS spectra were recorded on a MicroMass Q-TOF Micro spectrometer. Res-14-54. N-[2-(4-chlorophenyl)ethyl]-6-hydroxy- 7~niiro-3, 4-dihydroisoquinoline-

2(lH)-carbothioamide. ¹ H-NMR δ 2.95 (m, 4H), 3.84 (t, JN5.0 Hz, 2H), 3.94 (m, 2H), 4.89 (s, 2H), 5.55 (bs, IH), 7.00 (s, IH), 7.16 (d, J=8.3 Hz, 2H), 7.27 (d, J=8.3 Hz, 2H), 7.91 (s, IH), 10.5 (s, IH). ¹³ C-NMR δ 29.4, 34.7, 44.6, 47.0, 48.4, 118.9, 122.9, 126.0, 129.0, 129.0, 130.3, 130.3, 132.2, 133.7, 137.4, 146.7, 154.3, 181.9. ESI-MS calculated for C ₁₈ H ₁₉ ClN ₃ O ₃ S 392.0836 (M+H), found 392.0811.

Res-14-56. N-[2-(4-chlorophenyl)ethyl]-6-hydroxy-5-nitro-3,4-dihydroiso quinoline- 2(lH)-carbothioamide. ¹ H-NMR δ 2.97 (t, J=6.9 Hz, 2H), 3.26 (t, J=5.9 Hz, 2H), 3.76 (t, J=5.9 Hz, 2H), 3.96 (m, 2H), 4.90 (s, 2H), 5.51 (bs, IH), 7.08 (d, J=8.6 Hz, IH), 7.17 (d, J=8.4 Hz, 2H), 7.30 (m, 3H), 10.4 (s, IH). ¹³ C-NMR δ 27.5, 34.8, 44.2, 46.9, 49.2, 118.4, 127.2, 129.1, 129.1, 130.4, 130.4, 132.7, 133.1, 134.4, 135.0, 137.5, 154.1, 181.6. ESI-MS calculated for C ₁₈ H ₁₉ ClN ₃ O ₃ S 392.0836 (M+H), found 392.0860.

EXAMPLE 19. Synthesis of N-[2-(4-chlorophenyl)ethyl]-7,8-dihydro[l,3]dioxolo[4,5- g]isoquinoline-6(5H)-carbothioamide (Res-15-6) and 4,9-dichloro-N-[2-(4- chlorophenyl)ethyl]-7,8-dihydro[l,3]dioxolo[4,5-g]isoquinoli ne-6(5H)-carbothioamide (Res-15-11).

The title compounds were synthesized according to Scheme 12.

Boc anhydride/DMF

Scheme 12. Synthesis ofN-[2-(4-chlorophenyl)ethyl]-7,8-dihydro[l,3]dioxolo[4,5- g]isoquinoline-6(5H)-carbothioamide (Res-15-6) and 4,9-dichloro-N-[2-(4- 5 chlorophenyl)ethyl]-7,8-dihydro[l,3]dioxolo[4,5-g]isoquinoli ne-6(5H)-carbothioamide (Res-15-11).

To a suspension of 1 (1 eq.) in glacial acetic acid, SO ₂ Cl ₂ was added. After stirring for 0.5 h at r.t the solvents were removed under reduced pressure. The residual was repeatedly 0 suspended in toluene and evaporated affording 2. No further purification was done.

The corresponding amine (1 or 2) (1 eq.) dissolved in HBr (48 % in H ₂ O) was refluxed for 5h. The reaction solution was allowed to cool to r.t after which the solvent was removed under reduced pressure. The residual was repeatedly suspended in EtOAc and evaporated 5 yielding 3a and 3b (quantitative).

To a suspension of 3a or 3b (1 eq.) in H ₂ O, a solution of triethylamine (2.1 eq.), Boc ₂ O (1.05 eq.) in THF was added dropwise. After stirring at r.t overnight, the solution was concentrated, dissolved in EtOAc, washed with water, dried over Na ₂ SO ₄ , filtered and O evaporated. The product was purified by flash column chromatography (silica, Petroleum

Ether/EtOAc 1 :1) to give 4a (93%) or (silica, Petroleum Ether/EtOAc 5:1) to give 4b (52%)

A mixture of 4a or 4b (1 eq.) and NaOH (2.08 eq.) was added in small portions to a 130 5 °C solution of DMSO (5.3 ml/mmol 4) and CH ₂ Cl ₂ (O.lml/mmol 4) under N ₂ . After 2 Ii stirring at 130 ⁰ C ₅ NaOH (0.07 eq.) and CH ₂ Cl ₂ (0.02 ml/mmol 4) were added under N ₂ . After an additional hour the solution was cooled to r.t and separated between H ₂ O and EtOAc. The organic phase was washed repeatedly with water, dried (MgSO ₄ ), filtered and evaporated. The product was purified by flash column chromatography (silica, Petroleum O ether/EtOAc 3:1) giving 5a (86%) and (silica, Petroleum ether/EtOAc 5:1) giving 5b

(40%).

To a solution of 5a or 5b (1 eq.) in THF, HCOOH was added. The solution was heated at 60 ⁰ C for 7.5 h after which the solvent was removed under reduced pressure. The residue

was dissolved in anhydrous DMF and triethylamine (3 eq.) was added. The solution was stirred for 15 min. at r.t after which 2-(4-chlorophenethyl)isothiocyanate (1.2 eq.) was added. After stirring for 4 h at r.t the solution was separated between H ₂ O and EtOAc. The organic phase was washed repeatedly with water, washed with brine, dried over MgSO ₄ , filtered and evaporated. Flash column chromatography (silica, Petroleum ether/EtOAc 3:1) gave Res-15-6 (55%) and (silica, Petroleum ether/EtOAc 3:1) gave Res-15-11 (44%)

EXAMPLE 20. Physical data of the compounds of the invention General. ¹ H-NMR spectra and ¹³ C-NMR spectra were recorded with the following spectrometer, Bruker DRX-400 (at 400/100 MHz). CDCl ₃ (7.26/77.2 ppm) was used as solvent (calibration value shown in parenthesis). ESI-MS spectra were recorded on a MicroMass Q-TOF Micro spectrometer. Res-15-6. N-[2-(4-chlorophenyl)ethyl]-7, 8-dihydro[l, 3Jdioxolo[4, 5-g]isoquinoline-6(5H)- carbothioamide. ¹ H-NMR (CDCl ₃ 400 MHz) δ 2.82 (t, J=5.5 Hz, 2H), 2.96 (t, J=6.8 Hz, 2H), 3.80 (t, J=5.5 Hz, 2H), 3.95 (bs, 2H) 4.77 (s, 2H), 5.50 (bs, IH), 5.94 (s, 2H), 6.63 (s, IH), 6.65 (s, IH), 7.17 (d, J=8.2 Hz, 2H), 7.30 (d, J=8.2 Hz, 2H). ¹³ C-NMR (CDCl ₃ 100 MHz) δ 29.0, 34.9, 45.6, 46.9, 49.5, 101.2, 107.0, 108.2, 126.2, 128.6, 129.0, 129.0, 130.4, 130.4, 132.3, 137.6, 146,6, 147.0, 181.1. ESI-MS calculated for C ₁₉ H ₂₀ N ₂ O ₂ SCl (M+H) 375.0934, found 375.0922.

Res-15-11. 4, 9-dichloro-N-[2-(4-chlorophenyl)ethyl]-7, 8-dihydro[l, 3Jdioxolo[4, 5- g]isoquinoline-6(5H)-carbothioamide. ¹ H-NMR (CDCl ₃ 400 MHz) δ 2.87 (t, J=5.9 Hz, 2H), 2.97 (t, J=6.9 Hz, 2H), 3.98 (m, 4H), 4.69 (s, 2H), 5.60 (bs, IH), 6.13 (s, 2H), 7.18 (d, J=8.3 Hz, 2H), 7.29 (d, J=8.3 Hz, 2H). ¹³ C-NMR (CDCl ₃ 100 MHz) δ 25.7, 34.5, 44.8, 46.8, 46.8, 102.6, 110.8, 112.5, 124.7, 127.8, 128.9, 128.9, 130.2, 130.2, 132.5, 137.3, 143,6, 144.0, 181.8. ESI-MS calculated for C ₁₉ H ₁₈ N ₂ O ₂ SCl ₃ (M+H) 443.0155, found 443.0173.

EXAMPLE 21. Bronchorelaxation test

Apparatus and materials Dissection and mounting of lung tissue preparations. Lung tissue was obtained from patients undergoing lobectomia or pulmectomia due to lung carcinoma. The tissue was placed in a dissection chamber continuously perfused with 10 ml min ^'1 of a physiological saline solution (PSS) at room temperature. An airway was identified in the cut part of the lobe, and a bronchus of 10-20 mm length and 1-2 mm diameter was obtained. The bronchus was cut into rings of a width of about 2-3 mm. Each bronchial ring was cleaved to obtain an about rectangular oblong preparation, one end of which was tied to a small steel hook connected to a force transducer, while the other end of the preparation was attached to a fixed hook. This is followed by a period of adjustment, as described below. The preparation was mounted in an atmosphere containing 12% of oxygen and 6% of CO ₂ .

Experimental chamber. The experimental chamber has a volume of 5 ml. It is perfused with PSS at a rate of 3 ml min ^"1 . Two preparations are mounted in the chamber, and measurements on them are performed in parallel. For mechanical tensioning each force transducer (AME 801, SensoNor AJS, Horten, Norway) is connected to a

micrometer screw. The substances to be tested, the reference substance (capsazepine), and transmitter (LTD4) are injected upstream of the preparation(s).

Materials. PPS (physiological saline solution, in mM): NaCl, 117; KCl, 4.87; MgSO ₄ , 0.60; NaHCO ₃ , 25.0; CaCl ₂ , 1.60; glucose, 5.23. The solution is saturated with a mixture of 94% oxygen and 6% carbon dioxide, giving a pH of 7.40 ± 0.05 in the experimental chamber. AU substances are prepared as stock solution dissolved in the vehicles ethanol or DMSO. Leukotriene D4 (LTD4; Cayman Ltd.): 10 μl of a 100 μM ethanol stock solution. Capsazepine (Sigma Aldrich): 10 μl of a 0.1 M ethanol stock solution. Substance to be tested: 10-100 μl of a 0.01-0.1 M ethanol or DMSO stock solution. Solution for establishing the passive tension level: calcium-free PSS + 2 mM EGTA + 20 mM caffeine. To exclude effects by the test substance vehicle, ethanol or DMSO, respectively, were added during the entire experiment except during the presence of test substance.

Test procedure

An exemplary test is shown in the Figure in which capital letters indicate interference with the test system. The material for the preparation was a bronchus (inner diameter about 1 mm) from a male occasional smoker (41 yrs) but with the epithelium intact.

Adjustment and stretch. After mounting as described above the preparation is allowed to adjust with a low passive tone in the experimental chamber. The composition of the gas is changed to 94% (v/v) of oxygen. After a short adjustment period, PSS with

10 nM LTD4 is added to the experimental chamber upstream of the preparation (A). The preparation is stretched repeatedly (B) until it exerts a contraction force of around 150 mg. When the contraction has levelled off, leukotriene-free solution is administered for 1 hour (C), resulting in a relaxation. A second injection of 10 nM LTD4 (D) makes the preparation return to the tensioned state. At the peak tension leukotriene-free solution is again administered (E). After a third injection of 10 nM LTD4 (F) the preparation returns to the tensioned state. At the peak, PSS with 10 μM capsazepine (G) is added, resulting in a relaxation. After 1 h exposure to capsazepine, LTD4 is added, resulting in a contraction (H). In comparison with the control LTD4 contraction (F), a substantially weaker contraction is now observed (H). To obtain a measure of the test substance's bronchorelaxing effect the test and control forces registered in the experiment are compared. In the present experiment a remaining contraction (test force) of about 55 % of that caused by the control force was registered. After allowing one hour for return to baseline conditions (I) 10 nM LTD4 is again injected (J) to determine the reversibility of receptor inhibition. During steps C-F and I-J 10 μl ethanol per 100 ml PSS is present to compensate for potential vehicle effects. The experiment is concluded by adding calcium- free solution with addition of 2 mM EGTA and 20 mM caffeine for 20 min to establish the passive tension level (K). A bronchus tissue preparation is considered stable and thus fit for the evaluation of test substances if the difference in contraction between contractions D and F is less than 15 per cent.

The bronchorelaxing compounds according to the invention and some prior art compounds were tested for bronchorelaxation by substituting capsazepine in the test system, and were found to be active in this test system. A measure of the bronchorelaxing capacity of a candidate substance is obtained by comparing the result (% blocking of

contraction by LTD4) with that obtained with capsazepine. If the remaining contraction after exposure to a test substance is larger than after exposure to capsazepine, the test substance is less effective than capsazepine in regard of bronchorelaxing properties. If, on the other hand, the remaining contraction after exposure to a test substance is smaller than after exposure to capsazepine, the test substance is more effective than capsazepine in regard of bronchorelaxing properties.