CHIRAL TETRA-HYDRO BETA-CARBOLINE DERIVATIVES, APPLICATIONS THEREOF AS ANTIPARASITIC COMPOUNDS

The Invention relates to the use of chiral tetra-hydro β-carboline derivatives for the preparation of a pharmaceutical composition for the prevention and/or the treatment of parasitic diseases involving parasites having a phosphodiesterase activity. The invention also relates to some new chiral tetra-hydro β-carboline derivatives.

Malaria kills a child somewhere in the world every 30 seconds. It infects about 350-500 million people each year, causing approximately 1.3-3 million deaths annually (Campbell, Neil A. et a/. "Biology" Seventh edition. Menlo Park, CA: Addison Wesley Longman, Inc. 2005). Ninety per cent of those who die are in Africa, where malaria accounts for about one in five of all childhood deaths. The disease also contributes greatly to anaemia among children, a major cause of poor growth and development. During pregnancy, malaria is associated with maternal illness and severe anaemia. It contributes to low birth weight among newborn infants ~ one of the leading risk factors for infant mortality. Malaria also has serious economic impacts in Africa, slowing economic growth and development and perpetuating the vicious cycle of poverty. Malaria is caused by protozoan parasites of the genus

Plasmodium (phylum Apicomplexa): P. falciparum, P. malariae, P. ovale, and P. vivax. Their primary hosts and transmission vectors are female mosquitos of genus Anopheles; humans act as intermediate hosts. The P. falciparum variety of the parasite accounts for 80% of cases and 90% of deaths. Children under the age of five and pregnant women are the most vulnerable to the severe forms of malaria.

There are treatments and tools that can help. Sleeping under insecticide treated nets can reduce child mortality by 20 per cent. However, the insecticide loses its effectiveness over time, so the nets must be retreated. Prompt access to effective treatment can further reduce deaths. Intermittent preventive treatment of Malaria during pregnancy can significantly reduce the proportion of low birth weight infants and maternal

deaths.

Several drugs can be taken preventively. Generally, these drugs are taken daily or weekly, at a lower dose than would be used for treatment of a person who had actually contracted the disease. Use of prophylactic drugs is seldom practical for full-time residents of malaria- endemic areas, and their use is usually restricted to short-term visitors and travelers to malarial regions. This is due to the potentially high cost of purchasing the drugs, because long-term use of some drugs may have negative side effects, and because some effective anti-malarial drugs are difficult to obtain outside of wealthy nations.

Quinine was used starting in the seventeenth century as a prophylactic against malaria. The development of more effective alternatives such as quinacrine, chloroquine, and primaquine in the twentieth century reduced the reliance on quinine. Today, quinine is still used to treat chloroquine resistant Plasmodium falciparum, as well as severe and cerebral stages of malaria, but is not generally for malaria prophylaxis.

Modern drugs used preventively include mefloquine (Lariam®), doxycycline (available generically), and atovaquone proguanil hydrochloride (Malarone®). The choice of which drug to use is usually driven by what drugs the parasites in the area are resistant to, as well as side-effects and other considerations.

To be effective as a treatment in patients who have actually contracted malaria, these drugs have to be administered at daily or weekly higher dose. This has for inevitable consequences the increasing of negative side effects and to the development of an increased number of resistant to treatment parasites.

Sleeping sickness or African trypanosomiasis is a parasitic disease in people and animals, caused by protozoa of genus Trypanosoma and transmitted by the tsetse fly. The disease is endemic in certain regions of Sub-Saharan Africa, covering about 36 countries and 60 million people. It is estimated that 50,000 to 70,000 people are currently infected, the number having declined somewhat in recent years.

Trypanosoma brucei is parasitic protist species that causes African trypanosomiasis in humans and animals in Africa. There are 3 subspecies of T.brucei; T.b. brucei, T.b. gambiense and T.b. rhodesiense.

Pentamidine, a highly effective drug for the first stage of the disease, has been used since 1939. During the fifties, it was widely used as a prophylactic agent in Western Africa, leading to a sharp decline in infection rates. The organo-arsenical melarsoprol (Arsobal) was developed in the 1940s, and is effective for patients with second stage sleeping sickness. Eflomithine (difluoromethylomithine or DFMO), the most modem treatment, was developed in the 1970s and is today donated to the infected population. In 2005 a Phase III trial was testing the efficacy of the first oral treatment for Sleeping Sickness.

There is therefore an evident need for new effective drugs which could be used for the treatment and/or the prevention of malaria, and more generally all parasitic diseases due to tropical parasites like Plasmodium, Trypanosoma, Leishmania, at a lower daily or weekly dose than the drugs actually on the market and which could also be alternatively used to prevent and or treat parasitic diseases caused by resistant-to-treatment parasites. The inventors have developed the subject of the invention in order to remedy these problems.

In a paper published in Biochem. J., (2005, 392, 221-229), Yuasa, K. et al describe the biochemical features of one of four putative phosphodiesterases (PDE) encoded in the genome of P. falciparum, called PfPDEL On the basis of its inhibition profile, substrate preference and sequence, they suggest that PfPDEI could be successfully inhibited by inhibitors of human PDEs. However, they subsequently show that amongst 12 known inhibitors of human PDEs, only 4 actually weakly inhibit PfPDEI (micromolar range). Of these compounds, only zaprinast very weakly inhibits the growth of the parasite (35+/-4.2 μM*). Using site-directed mutagenesis, they demonstrate that the aminoacid residues concerned with the binding of zaprinast differ between mammalian PDE5 and PfPDEI . They conclude that

Structure-Activity-Relationship for potential PfPDEI inhibitors are expected to significantly differ from the one established in the family of PDE5 inhibitors.

The tadalafil, a marketed human PDE5 inhibitor, is totally inactive on parasite cultures. The authors discovered that adding a lateral chain of selected structure on close tadalafil analogues generate very potent inhibitors of the growth of parasites like Plasmodium falciparum and Trypanosoma brucei (nanomolar range).

In particular, the Inventors have found that judiciously selected tetra-hydro β-carboline type compounds, responding to hereafter defined formula (I), have an enhanced antiparasitic activity against the genus Plasmodium, and more generally against all parasites having a phosphodiesterase (PDE) activity (Trypanosoma, Leishmania...), and can therefore advantageously be used, as an active principle, for the prevention and/or the treatment of parasitic diseases involving parasites having a PDE activity.

The compounds responding to hereafter defined formula (I) can therefore also be used, as an active principle, for the preparation of an antiparasitic pharmaceutical composition for the prevention and/or the treatment of parasitic diseases involving parasites having a PDE activity. A first subject of the Invention is therefore the use, as an active principle, of at least one chiral tetra-hydro β-carboline derivative of general formula (I) below:

or a pharmaceutically acceptable salt thereof, in which: - Ri and R ₂ , identical or different, represent a hydrogen atom or a fluorine atom;

- k is an integer equal to 0 or 1 ;

- R ₃ is selected from the group consisting of:

■ a phenyl ring optionally substituted by a methylenedioxy radical, a difluoromethylenedioxy radical attached to the phenyl ring at the 3'- 4' carbon atoms, a thiadiazole fused with the phenyl ring at the 3'-4' positions, a difluoromethoxy radical or a chlorine atom in the para 4' position with regard to the attachment point of said phenyl ring to the piperidine cycle directly bearing R ₃ when k = 0 or to the carbon atom of the -CH ₂ - group when k = 1 , a methyloxy radical and/or a hydroxyl radical in the meta 3' position and/or in the para 4' position with regard to the attachment point of said phenyl ring to the piperidine cycle directly bearing R ₃ when k = 0 or to the carbon atom of the -CH ₂ - group when k = 1 ;

^■ a 3'-λ/-pyridine ring optionally substituted by a substituent chosen from a chlorine atom, a fluorine atom, a methoxy group, and a methylthio group, said substituent being in the 4' position with regard to the attachment point of said 3'-A/-pyridine ring to the piperidine cycle directly bearing R ₃ when k = 0 or to the carbon atom of the -CH ₂ - group when k = 1 ;

- R ₄ is a group selected in the group consisting of the following groups: -NH-A-R ₅ ; -NHC(O)-R ₅ and the groups of formulas (ll-a) to (ll-c) below:

(ll-a) (||-b) (ll-c) in which:

■ n is an integer equal to 0 or 1 ,

^■ h is an integer equal to 0, 1 , 2 or 3, ^■ m is an integer equal to 0, 1 or 2,

■ the arrows represent the attachment point of groups of formulas (ll-a), (ll-b) or (ll-c) on the formula (I) derivative piperazine-dione cycle;

■ Xi is a carbon or a nitrogen atom;

- A represents a single bond or a Ci to C ₃ linear or branched alkyl chain optionally interrupted or terminated by an oxygen atom;

- R ₅ is selected in the group consisting of: i) a group of formula (III) below:

in which the arrow represents the attachment point of said group of formula (III) to A via a covalent bond; ii) a group of formulas (IV) or (V) below:

(IV) (V) in which the arrows represent the attachment point of said group of formulas (IV) and (V) to A via a covalent bond and R ₆ and R ₇ , identical or different, represent a Ci to C ₄ alkyl group; and iii) a group of the following formula (Vl):

in which:

- the arrow represents the attachment point of said group of formula (Vl) to A via a covalent bond,

- Y-] and Y ₅ , identical or different, represent -CH=; -N= or -C(R ₈ )= in which Rs represents an hydrogen atom, a methyl radical or a halogen atom selected from chlorine and fluorine;

- Y ₃ represents -CH=; -N=; -C(R ₉ )= or -C(R ₁₀ )=;

- Y ₂ and Y ₄ , identical or different, represent -CH=, -N=, -C(R ₉ )= or -C(R ₁₀ )=;

- R ₉ represents a hydrogen atom or a group chosen from a

methyl, an amino group and a methyloxy radical;

- Y ₂ and Y ₃ can also form together a methylenedioxy group;

- R-io represents a hydrogen atom or a solubilizing group; wherein the asymmetric carbon atom (number 12a) of said derivative of formula (I) is in the R conformation, for the preparation of a pharmaceutical composition for the prevention and/or the treatment of parasitic diseases involving parasites having a phosphodiesterase (PDE) activity.

The inventors have actually demonstrated that the compounds of formula (I) according to the invention inhibit Plasmodium growth in infected mice at doses ranging from 1 to 50 mg per kg when administered by the oral route {per os) or by intra-peritoneal or intravenous injections. They also inhibit the growth of Plasmodium parasites in red blood cells at concentrations ranging from 1 nM to 10 μM.

The chiral tetra-hydro β-carboline derivatives of formula (I) make possible to prevent and/or treat parasitic diseases involving parasites which have developed a particular resistance to usual antiparasitic agents, when they are administered alone or in combination with usual antimalarial drugs.

According to a prefered embodiment of the invention, the chiral tetra-hydro β-carboline derivatives of formula (I) are used for the preparation of a pharmaceutical composition for the prevention and/or the treatment of malaria, leishmaniasis or trypanosomiasis; the prevention and treatment of malaria being the most prefered indication.

In the derivatives of formula (I), the solubilizing groups above- mentioned for R ₁₀ can be chosen amongst the prior well known water solubilizing groups, preferably such as those of the following formula (S):

in which: - p is an integer equal to 1 , 2, 3, or 4;

- B is (CH ₂ )q-Rii wherein q is an integer equal to 0 or 1 and R-i ₁ is selected from the group consisting of an amino group, a morpholine ring, a 4-methyl-piperazinyl group, a piperidinyl group, a methyloxy radical and a hydroxyl radical;

- the arrow represents the attachment point of said solubilizing group to the carbon atom designated for Y ₂ , Y ₃ and Y ₄ in the corresponding group of formula (Vl) as above-defined.

According to a preferred embodiment of the invention, R ₃ represents one of the following groups:

According to a preferred embodiment of the invention, the solubilizing group of formula (S) represents one of the following groups of formulas (S-1 ) to (S-8):

(S-6) (S" ⁷ ) (S-8)

According to another preferred embodiment of the invention, R ₄ is a group of formula (ll-b) in which h = 1 , n = 1 and X ₁ is a carbon or a nitrogen atom.

According to a preferred embodiment of the invention R ₄ is a group of formula (ll-b) in which h = 1, n = 0 and X ₁ is a carbon or a nitrogen atom.

According to a preferred embodiment of the invention R ₄ is a group of formula (ll-b) in which h = 0, n = 0 and Xi is a carbon or a nitrogen atom.

In the groups -NH-A-R ₅ and in the groups of formulas (ll-a) to (ll-c) mentioned for R ₄ , A preferably represents -CH ₂ -, -CH(CH ₃ )-, -C(CHs) ₂ -, -CH ₂ -CH ₂ -, -C(CHa) ₂ -CH ₂ - or -CH ₂ -CH ₂ -O-.

In the groups of formula (ll-a) to (ll-c), R ₅ preferably represents one of the following groups of formulas (IV-1 ) to (IV-34):

(IV-1 ) (IV-2) (IV-3) (IV-4)

(IV-15 )

(IV-19)

(IV-20)

(IV-21 ) (IV-22)

(IV-23) (IV-24)

(IV-33)

(IV-32) and

(IV-34) Amongst said derivatives of formula (I), those in which: i) k = 0, R ₃ represents a piperonyl, a benzothiadiazole, a

4-chloro-3-N-pyridine or a 4-fluoro-3-N-pyridine group and R ₄ represents a group of formula (ll-b) in which Xi is a carbon atom or a nitrogen atom, h = 1 , n = 1 , A is -CH ₂ - and R ₅ is a group of formulas (IV-2), (IV-3), (IV-8), (IV-10) or (IV-12); ii) k = 0, R ₃ represents a piperonyl, a benzothiadiazole, a phenyl, a 4-chloro-3-N-pyridine group, a 3,4-dimethyloxyphenyl, 3,4- dihydroxyphenyl, 3-hydroxy-4-methyloxy-phenyl, 4-hydroxy-3-methyloxy- phenyl or a difluoromethylenedioxy radical attached to the phenyl ring at the 3'-4' carbon atoms and R ₄ represents a group of formula (ll-b) in which Xi is a carbon atom or a nitrogen atom, h = 1 , n = 0, A is -CH ₂ - and R ₅ is a group of formula (IV-1 ) or (IV-2); iii) k = 0, R ₃ represents a phenyl group and R ₄ represents a group of formula (ll-b) in which Xi is a carbon atom or a nitrogen atom, h = 1 , n = 1 , A is -CH ₂ - and R ₅ is a phenyl group of formula (IV-1 ); iv) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-c) in which X ₁ is a carbon atom

or a nitrogen atom, m = 3, A is -CH ₂ - and R ₅ is a phenyl group of formula

(IV-1 ); v) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-c) in which X ₁ is a carbon atom or a nitrogen atom, m = 2, A is -CH ₂ - and R ₅ is a phenyl group of formula

(IV-1 ); vi) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-b) in which X ₁ is a carbon atom or a nitrogen atom, h = 1 , n = 1 , A is -CH ₂ - and R ₅ is a phenyl group of formula (IV-1); vii) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-b) in which Xi is a carbon atom or a nitrogen atom, h = 1 , n = 0, A is -CH ₂ - and R ₅ is a group of formulas

(IV-4), (IV-8), (IV-10), (IV-12), (IV-13), (IV-14) or (IV-34); viii) k = 0, R ₃ represents a 3',4'-(difluoromethylenedioxy)- phenyl group, R ₄ represents a group of formula (ll-b) in which X ₁ is a carbon atom or a nitrogen atom, h = 1 , n = 1 , A is -CH ₂ - and R ₅ is a group of formula

(IV-1 ); ix) k = 0, R ₃ represents a 4'-difluoromethoxyphenyl group, R ₄ represents a group of formula (ll-b) in which X-i is a carbon atom or a nitrogen atom, h = 1 , n = 1 , A is -CH ₂ - and R ₅ is a group of formula (IV-1 ); x) k = 1 , R ₃ represents a phenyl group, R ₄ represents a group of formula (ll-b) in which Xi is a carbon atom or a nitrogen atom, h = 1 , n = 1 ,

A is -CH ₂ - and R ₅ is a group of formula (IV-1 ) or (IV-2); xi) k = 0, R ₃ represents a piperonyl, a benzothiadiazole, a

4-fluoro-3-N-pyridine or a 4-chloro-3-N-pyridine group, R ₄ represents a group of formula (ll-b) in which X ₁ is a carbon atom or a nitrogen atom, h = 1 , n = 1 ,

A is single bond and R ₅ is a group of formula (IV-9), xii ) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group, R ₄ represents a group of formula (ll-b) in which X ₁ is a carbon atom or a nitrogen atom, h = 1 , n = 0, A is -CH ₂ - and R ₅ is a group of formula (IV-15),

(IV-16), (IV-17), (IV-19), (IV-20), (IV-23), (IV-24), (IV-26), (IV-25), (IV-27),

(IV-28), (IV-30), (IV-33) or (IV-32); xiii) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group, R ₄ represents a group of formula (ll-b) in which Xi is a carbon atom or a nitrogen atom, h = 0, n = 0, A is -CH ₂ - and R ₅ is a group of formula (IV-26), (IV-25) or (IV-27); xiv) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-b) in which Xi is a carbon atom or a nitrogen atom, h = 1 , n = 0, A is -C(CH ₃ ) ₂ - and R ₅ is a phenyl group of formula (IV-1); xv) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-b) in which X ₁ is a carbon atom or a nitrogen atom, h = 1 , n = 0, A is -CH ₂ -CH ₂ - and R ₅ is a phenyl group of formula (IV-1); xvi) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-b) in which X ₁ is a carbon atom or a nitrogen atom, h = 1 , n = 0, A is -CH ₂ -CH ₂ -O- and R ₅ is a phenyl group of formula (IV-1); xvii) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-b) in which X ₁ is a carbon atom or a nitrogen atom, h = 1 , n = 0, A is -C(CH ₃ ) ₂ -CH ₂ - and R ₅ is a phenyl group of formula (IV-1 ); xviii) k = 0, R ₃ represents a piperonyl or a benzothiadiazole group and R ₄ represents a group of formula (ll-b) in which Xi is a carbon atom or a nitrogen atom, h = 1 , n = 0, A is -CH(CH ₃ ) and R ₅ is a phenyl group of formula (IV-1 ); and pharmaceutically acceptable salts thereof, are preferred.

Among compounds of formula (I) here above listed under items i) to xviii), those in which Ri and R ₂ simultaneously represent a hydrogen atom and those in which one of Ri and R ₂ represents a fluorine atom while the other of R ₁ and R ₂ represents a hydrogen atom are particularly preferred.

Among compounds of formula (I) here above listed under

items i) to xviii), those in which Xi represents a carbon atoms are also particularly prefered.

As particular compounds of formula (I), one can cite: (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(1-methyl-1H-imidazol-2-yl methyO-pyrrolidin-S-yO^.S.θJ.^.^a-hexahydro-pyrazinoCr^'il .ejpyridofSλ- ιb]indole-1 ,4-dione;

(6S,12aR)-6-Benzo[1 _! 3]dioxol-5-yl-2-[(R)-1-(1-methyl-1H-imidazol-2-yl methyO-pyrrolidin-S-yl^.S.ej.^.^a-hexahydro-pyrazinotr^'ii ^jpyridotS^- b]indole-1 ,4-dione; - (6R,12aR)-2-(1 -Benzyl-piperidin-4-yl)-6-phenyl-2, 3,6,7,12,12a-hexahydro- pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(6-chloro-pyrid in-3-yl)-2,3,6,7, 12,12a-hexahydro-pyrazino[1 ^l ,2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6S,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(6-chloro-py ridin-3-yl)-2,3,6,7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6-(6-chloro-pyridin -3-yl)-2,3,6,7,12, 12a-hexahydro-pyrazino[1 \2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[3-(benzyl-methyl-amino)-propyl]-2,3,6, 7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-/5]indole-1 ,4-dione; - (6S,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[3-(benzyl-methyl-amino)-propyl]-2,3,6, 7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[2-(benzyl-methyl-amino)-ethyl]-2,3,6,7, 12, 12a-hexahydro-pyrazino[1 ^l ,2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

- (6S,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[2-(benzyl-methyl-amino)-ethyl]-2,3,6,7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-ib]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-(1-benzyl-piperidin-4-yl)-2,3,6,7,12,12a- hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-/?]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-(1-benzyl-piperidin-4-yl)-9-fluoro-2,3,6, 7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-(1-benzyl-piperidin-4-yl)-9-fluoro-

2,3,6,7, 12, 12a-hexahydro-pyrazino[1',2 ^l :1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-benzo[1 ,3]dioxol-5-ylmethyl-

pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro-pyrazino[1',2': 1 ,6]pyrido [3,4-b]indole -1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(4-methoxy-benzyl)-pyrrolidin-3- yO^.S.βJ.^.^a-hexahydro-pyrazinoti'^'il .eipyridotS^-^indole-I λ-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-pyridin-2-ylmethyl-pyrrolidin-3- yl)-2,3,6,7,12,12a-hexahydro-pyrazino[r,2':1 ,6]pyrido [3,4-/}]indole-1 ,4-dione; (6R, 12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1 -(4-dimethyl-amino-benzyl)- pyrrolidin-3-yl]-2 _I 3 _J 6,7,12 _> 12a-hexahydro-pyrazino[1 ^I ,2 ^I :1 ,6]pyrido[3,4-b]indole-

1 ,4-dione; - 2-[(R)-3-((6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-1 ,4-dioxo-3,4,6,7,12,12a-hexa hydro-IH-pyrazinofi'^M .eipyridop^-^indol^-yO-pyrrolidin-i-y^-ethyl}- carbamic acid ferf-butyl ester;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1 -benzyl-pyrrolidin-3-yl)-2,3,6,7,12, 12a-hexahydro-pyrazino[1 ^l _> 2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione; - (6S,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-benzyl-pyrrolidin-3-yi)-2,3,6,7,12, 12a-hexahydro-pyrazino[1 ',2': 1 ,6]pyrido[3,4-ib]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[1-(1-methyl-1H-imidazol-2-ylmethyI)- piperidin^-y^^.ej.^.^a-hexahydro-pyrazinoti'^'ii .ejpyridofS^-^indole- 1 ,4-dione; - (6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6-(4-difluoromethoxy-p henyl)-2,3 _> 6,7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6-(2 _I 2-difluoro-benzo[1 ,3]dioxol-5-yl)-2, 3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,63pyrido[3,4-/?]indole-1 ,4-dione;

6,7,12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(δR.^aR^β-Benzyl^i-benzyl-piperidin^-yl^.S.ej.^.^a-hexahyd ro- pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; - (6S,12aR)-6-Benzyl-2-(1-benzyl-piperidin-4-yl)-2,3,6,7,12,12 a-hexahydro- pyrazino[1 ,4-dione;

- (6R,12aR)-6-Benzyl-2-[1 -(1 -methyl-1 H-imidazol-2-ylmethyl)-piperidin-4-yl]-2,

3,6 _I 7,12,12a-hexahydro-pyrazino[1 ^l ,2 ^l :1 ,6]pyrido[3,4-ib]indole-1 ₎ 4-dione;

- (6S,12aR)-6-BenzyI-2-[1-(1-methyI-1H-imidazol-2-ylmethyl)-pi peridin-4-yl]-2, 3,6,7,12, 12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6R, 12aR)-2-{(R)-1 -[4-(2-{2-[2-(2-Amino-ethoxy)-ethoxy]-ethoxy}-ethoxy)- benzyl]-pyrrolidin-3-yl}-6-benzo[1 ,3]dioxol-5-yl-2,3,6,7,12,12a-hexahydro- pyrazino[1 ',2':1 ,6]pyrido[3,4-£>]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[3-(2-dimethylamino-ethoxy)- benzyO-pyrrolidin-S-yl^^^J.^.^a-hexahydro-pyrazinotr^M .θjpyridop^- 6]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(2-dimethylamino-ethoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4- jb]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[3-(3-morpholin-4-yl-propoxy)- benzyl3-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4- yb]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-{4-[3-(4-methyl-piperazin-1-yl)- propoxy]-benzyl}-pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[3-(2-morpholin-4-yl-ethoxy)- benzyl]-pyrrolidin-3-yl}-2, 3,6,7,12,12a-hexahydro-pyrazino[1\2':1 ,6]pyrido[3,4- b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(2-morpholin-4-yl-ethoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4- jb]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(3-morphoIin-4-yl-propoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ^l ,2':1 ,6]pyrido[3,4- jb]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(3-piperidin-1-yl-propoxy)- benzyπ-pyrrolidin-S-ylJ^.S.δJ.^.^a-hexahydro-pyrazinofi'^' il .δjpyridoP^- i?]indole-1 ,4-dione;

(6R ₁ 12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(3-dimethylamino-propoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 \2':1 ,6]pyrido[3,4- b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[3-(2-methoxy-eth oxy)-benzyl]- pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4- b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(2-methoxy-ethoxy)-benzyl]- £>]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[3,4-bis-(2-methoxy-ethoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12 _> 12a-hexahydro-pyrazino[1 ^l _l 2':1 ,6]pyrido[3,4- b]indole-1 ,4-dione;

- (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(2,2-difluoro-b enzo[1,3]dioxol-5- yl)-2,3,6,7, 12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-/?]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[1-(1-methyl-1H-imidazol-2-ylmethyl)- piperidin^-yO^.S.ej.^.^a-hexahydro-pyrazinofr^'ii .eipyridotS^-blindole- 1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((S)-1-benzyl-pyrrolidin-3-yl)- ,4-dione; - (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(3,4-dimethoxy- phenyl)-

2,3,6,7,12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-phenyl-2,3,6 ,7, 12,12a- hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-benzyl-pyrro!idin-3-yl)-9-fluoro- 2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-)b]indole-1 ,4-dione;

(6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(3,4-dihydroxy- phenyl)- 2,3,6,7,12,12a-hexahydro-pyrazino[1 ^l ,2':1 _I 6]pyrido[3,4-/3]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(2-methyl-benzyl)-pyrrolidin-3-yl]- 2,3,6,7,12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(2-chIoro-benzyl)-pyrrolidin-3-yl]- 2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R _I 12aR)-6-Benzo[1 _> 3]dioxol-5-yl-2-[(R)-1-(2,6-dichloro-benzyl)-pyrrolidin-3-

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(2,4-difluoro-benzyl)-pyrrolidin-3- yl^.S.ej.^.^a-hexahydro-pyrazinotr^'il .eipyridotS^-^indole-i ^-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-benzyl-pyrrolidin-3-yl)-10-fluoro- 2,3,6,7,12,12a-hexahydro-pyrazino[r,2':1 ,6]pyrido[3,4-ib]indole-1 ,4-dione;

- (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(3-hydroxy-4-me thoxy-phenyl)- 2,3,6 _> 7,12,12a-hexahydro-pyrazino[1 ^l ,2':1 _I 6]pyrido[3,4-/3]indole-1 _> 4-dione;

- (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(4-hydroxy-3-me thoxy-phenyl)- 2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-ib]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,2,5]thiadiazol-5-yl-2-{(R)-1 -[3-(2-methoxy-ethoxy)- benzylj-pyrrolidin-S-yl^.S.δJ.^.^a-hexahydro-pyrazinoti'^M .eipyridotS^- b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,2,5]thiadiazol-5-yl-2-{(R)-1-[3,4-bis-(2-methoxy- ethoxy^benzyO-pyrrolidin-S-yl^^^J.^.^a-hexahydro- pyrazino[1',2':1 ,6]pyrido[3 _l 4-jfcι]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,2,5]thiadiazol-5-yl-2-{(R)-1-[4-(2-methoxy-ethoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12 _> 12a-hexahydro-pyrazino[1 ^I ,2':1 ,6]pyrido[3,4- fo]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(1-methyl-1-phenyl-ethyl)- pyrrolidin-S-yl^.S^J.^.^a-hexahydro-pyrazinoli'^M .δlpyridotS^-^indole- 1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(1-phenyl-ethyl)-pyrrolidin-3-yl]- 2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-phenethyl-pyrrolidin-3-yl)- ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxoI-5-yl-2-[(R)-1-(2-phenoxy-ethyl)-pyrrolidin-3-yl]- 2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-6-(3a,7a-Dihydro-benzo[1 ,3]dioxol-5-yl)-2-[(R)-1-(1 ,1-dimethyl-2- phenyl-ethyl)-pyrrolidin-3-yl]-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

(6R,12aR)-6-(3a,7a-Dihydro-benzo[1 ,3]dioxol-5-yl)-2-{1-[3-(2-methoxy- ethoxy)-benzyl]-azetidin-3-yl}-2, 3,6,7,12, 12a-hexahydro- pyrazino[1 \2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-2-{1-[3,4-Bis-(2-methoxy-ethoxy)-benzyl]-azetidin- 3-yr}-6~(3a,7a- dihydro-benzo[1 ,3]dioxol-5-yl)-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; and pharmaceutically acceptable salts thereof.

As used herein, the term "optionally" means that the subsequently described event(s) may or may not occur, and include(s) both event(s), which occur, and events that do not occur.

"Amino" includes moieties of the formula NR- ₁₂ R ₁₃ wherein R ₁₂ and R- ₁₃ are each independently hydrogen or a methyl.

As used herein, the terms "group", "residue" and"radical" or "groups", "residues" and "radicals" are usually used as synonyms, respectively as it is common practice in the art.

As used herein, the term "disease" means a disease caused by infection with a parasite. More preferably, the disease is caused by infection with a parasite of genus Plasmodium, Trypanosoma and Leishmania.

Certain of the compounds described herein may contain one or more chiral atoms, or may otherwise be capable of existing as in two or more stereoisomers, which are usually diastereomers. Accordingly, the compounds of this invention include mixtures of stereoisomers, especially as well as purified stereoisomers, or stereoisomerically enriched mixtures. Also included within the scope of the invention are any wholly or partially equilibrated mixtures thereof. The present invention also covers the individual isomers of the compounds represented by the formula above as mixtures with isomers thereof in which one or more chiral centers are inverted. Also, it is understood that all tautomers and mixtures of tautomers of the compounds of the present invention are include within the formulae and subformulae corresponding thereto.

"Pharmaceutically acceptable salts" refers to any conventional acid-addition salts or base-addition salts that retain the biological

effectiveness and properties of the chiral tetra-hydro β-carboline derivatives of formula (I) and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases. Sample acid-addition can be acid addition salts formed with pharmaceutically acceptable acids. Such a definition encompasses hydrochloride, hydrobromide, sulphate or bisulphate, phosphate or hydrogenophosphate, acetate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, gluconate, methanesulphonate, benzene-sulphonate and paratoluene-sulphonate salts of tetra-hydro β-carboline derivates of formula (I). Sample base-addition salts include those derived from ammonium, potassium, sodium and quaternary ammonium, potassium, sodium and quaternary ammonium hydroides, such as for example, tetramethylammoniumhydroxide. The chemical modification of a pharmaceutical compound (i.e drug) into a salt is a technique well known to pharmaceutical chemists to obtain improved physical and chemical stability, hygroscopicity, flowability and solubility of the compounds.

The pharmaceutical composition of the present invention may be administered by any suitable route, for example, by oral, buccal, inhalation, sublingual, nasal, percutaneous, i.e., transdermal, or parenteral (including intravenous, intramuscular, subcutaneous, intraperitoneal, and intracoronary) administration. Therefore, the pharmaceutical composition of the invention can be provided in various forms, such as in the form of hard gelatin capsules, of capsules, of compressed tablets, of suspensions to be taken orally, of lozenges or of injectable solutions or in any other form appropriate to the method of administration. According to a prefered embodiment of the invention, the pharmaceutical composition is for an oral administration.

The pharmaceutical composition according to the invention include those wherein the at least one chiral tetra-hydro β-carboline derivative of above-defined formula (I) is administered in an effective amount to achieve its intended purpose. Determination of the effective amounts is well within the capability of those skilled in the art.

A "therapeutically effective dose" refers to that amount of the

chiral tetra-hydro β-carboline derivatives of formula (I) that results in achieving the desired effect. Toxicity and therapeutic efficacy of such derivatives can be easily determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD ₅₀ (the dose lethal to 50% of the population) end the ED ₅₀ (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, which is expressed as the ratio between LD ₅₀ and ED ₅₀ . β-carboline derivatives of formula (I) which exhibit high therapeutic indices are of course preferred. The data obtained from such data can be used in formulating range of dosage for use in humans. The dosage of β-carboline derivatives of formula (I) preferably lies within a range of circulating concentrations that include the ED ₅₀ with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed, and the route of administration. The exact formulation, route of administration, and dosage can be chosen by the individual physician in view of the patient's conditions. Dosage amount and interval of administration can be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the preventive or therapeutic effects. The amount of pharmaceutical composition administered will therefore depend on the subject being treated, on the subject's weight, the severity of the affliction and the manner of administration.

For human use, the chiral tetra-hydro β-carboline derivatives of formula (I) can be administered alone, but they are preferably administered in admixture with at least one pharmaceutically acceptable carrier, the nature of which will depend on the intended route of administration and the presentation form. Pharmaceutical composition for use according to the present invention thus can be formulated in a conventional manner using one or more physiologically acceptable carriers comprising one or more excipients and auxiliaries that facilitate processing of the chiral tetra-hydro β-carboline derivatives of formula (I) into preparations which can be used pharmaceutically. Amongst the excipients and auxiliaries which can be used

in the pharmaceutical composition according to the invention, one can mention antiagglomerating agents, antioxidants, preservatives agents, dyes, vitamins, inorganic salts, taste-modifying agents, smoothing agents, coating agents, isolating agents, stabilizing agents, wetting agents, anti-caking agents, dispersing agents, emulsifying agents, aromas, penetrating agents, solubilizing agents, etc., mixtures thereof and generally any excipient conventionally used in the pharmaceutical industry.

By way of example, when the pharmaceutical composition is administered orally, the carrier may comprise one or several excipients such as talc, lactose, starch or modified starches, cellulose or cellulose derivatives, polyethylene glycols, acrylic acid polymers, gelatin, magnesium stearate, animal or vegetal fats of natural or synthetic origin, paraffin derivatives, glycols, etc....

In addition to the at least one chiral tetra-hydro β-carboline derivative of formula (I), the pharmaceutical composition may also comprises one or more additional usually used as antiparasitic drugs. These antiparasitic drugs are drugs well known to a person skilled in the art. Without limitations, these antiparasitic drug can be chosen from eflomithine, melarsoprol, pentamidine isothionate, suramin, mifurtimox, sodium stibogluconate, meglumine antimoniate, amphotericin B, pentamidin, paromomycin, miltefosin, artemisin and its derivatives, sulfadoxine-pyrimethamine, artemether, lumefantine, amodiaquine, chloroquine, quinacrine and primaquine.

For general information about the formulation and administration of pharmaceutical compositions, one can obviously refer to the book "Remington's Pharmaceutical Sciences", last edition. Of course, a person skilled in the art will take care on this occasion that the excipient(s) and/or auxiliary(ies) optionally used are compatible with the intrinsic properties attached to the pharmaceutical composition in accordance with the invention.

These pharmaceutical compositions can be manufactured in a conventional manner, e.g., by conventional mixing, dissolving, granulating, dragee-making, emulsifying, encapsulating, entrapping, or lyophilizing processes. Proper formulation is dependent upon the route of administration

chosen.

Amongst the chiral tetra-hydro β-carboline derivatives of the above-defined formula (I), some compounds have never been disclosed in the literature.

A further subject of the present invention is therefore the chiral tetra-hydro β-carboline derivatives of formula (I ¹ ) below:

or a pharmaceutically acceptable salt thereof, in which:

- R'i and R' _2j identical or different, represent a hydrogen atom or a fluorine atom;

- k' is an integer equal to 0 or 1 ;

- R' ₃ is selected from the group consisting of:

^■ a phenyl ring optionally substituted by a methylenedioxy radical, a difluoromethylenedioxy radical attached to the phenyl ring at the 3'-4' carbon atoms, a thiadiazole fused with the phenyl ring at the 3'-4' positions, a difluoromethoxy radical or a chlorine atom in the para 4' position with regard to the attachment point of said phenyl ring to the piperidine cycle directly bearing R' ₃ when k' = 0 or to the carbon atom of the -CH ₂ - group when k ¹ = 1 , a methyloxy radical and/or an hydroxyl radical in the meta 3' position and/or in the para 4' position with regard to the attachment point of said phenyl ring to the piperidine cycle directly bearing R' ₃ when k' = 0 or to the carbon atom of the -CH ₂ - group when k' = 1 ;

^■ a 3'-λ/-pyridine ring optionally substituted by a substituent chosen from a chlorine atom, a fluorine atom, a methoxy group, and a methylthio group, said substituent being in the 4' position with regard to the attachment point of said 3'-/V-pyridine ring to the piperidine cycle directly

bearing R' ₃ when k' = 0 or to the carbon atom of the -CH ₂ - group when k ¹ = 1 ;

- R ₄ is a group selected in the group consisting of the following groups: -NH-A-R' ₅ ; -NHC(O)-R' ₅ and the groups of formulas (ll'-a) to (ll'-c) below:

(ll'-a) (ll'-b) (ll'-c) in which:

■ n' is an integer equal to 0 or 1 ,

^■ h' is an integer equal to 0 or 1 , 2 or 3,

^■ m' is an integer equal to 0, 1 or 2, ^■ the arrows represent the attachment point of groups of formulas (ll'-a), (ll'-b) or (ll'-c) on the formula (I ¹ ) derivative piperazine-dione cycle;

^■ X'-i is a carbon or a nitrogen atom;

- A' represents a single bond or a Ci to C ₃ linear or branched alkyl chain optionally interrupted or terminated by an oxygen atom;

- R' ₅ is selected in the group consisting of: i) a group of formula (III ¹ ) below:

in which the arrow represents the attachment point of said group of formula (III ¹ ) to A' via a covalent bond; ii) a group of formulas (IV) or (V) below:

(IV) (V)

in which the arrows represent the attachment point of said group of formula (IV) and (V) to A' via a covalent bond and R' ₆ and R' ₇ , identical or different, represent a Ci to C ₄ alkyl group; and iii) a group of the following formula (Vl'):

in which:

- the arrow represents the attachment point of said group of formula (Vl') to A' via a covalent bond,

- V ₁ and V ₅ , identical or different, represent -CH=; -N= or -C(R' ₈ )= in which R' ₈ represents an hydrogen atom, a methyl radical or a halogen atom selected from chlorine and fluorine;

- V ₃ represents -CH=; -N=; -C(R' ₉ )= or -C(R' ₁₀ )=;

- V ₂ and V ₄ , identical or different, represent -CH=, -N=, -C(Ry= Or -C(R'^=; - R'g represents a hydrogen atom or a group chosen from a methyl, an amino group and a methyloxy radical;

- V ₂ and V ₃ can also form together a methylenedioxy group;

- R'io represents a hydrogen atom or a solubilizing group; wherein the asymmetric carbon atom number 12a of said derivatives of formula (I ¹ ) is in the R conformation; with the provisos that:

^* when k' = 0, and R'i and R ₂ simultaneously represent a hydrogen atom, and R' ₃ is a piperonyl group, and R' ₄ represents a group of formula (ll'-b) wherein h ¹ = 1 , n' = 0, X'i is a carbon or a nitrogen atom, and A' = -CH ₂ -, then R' ₅ is different from a phenyl group, a pyridine group, a pyrimidine group, an imidazole group, a pyrazine group and a 1-methylimidazole group;

* when k' = 0, and R'I and R ₂ simultaneously represent a hydrogen atom, and R' ₃ is a piperonyl group, and R ₄ represents a group of formula (ll'-b) wherein h' = n ¹ = 0, X'i is a carbon or a nitrogen atom, and

A' = -CH ₂ -, then R' ₅ is different from a phenyl group;

^* when k' = 0, and R'i and R' ₂ simultaneously represent a hydrogen atom, and R'z is a piperonyl group, and R' ₄ represents a group of formula (ll'-b) wherein h' = n' = 1 , X'i is a carbon or a nitrogen atom, and A' = -CH ₂ -, then R' ₅ is different from a phenyl group.

The compounds excluded from the above-defined formula (I ¹ ) by these three provisos are known in the art, some of them being disclosed by Graham N. Maw et al., Bioorganic & Medicinal Chemistry Letters 13 (2003) 1425-1428. In the derivatives of formula (I ¹ ), the solubilizing groups above- mentioned for R'-io can be chosen amongst the prior well known water solubilizing groups, preferably such as those of the following formula (S'):

in which: - p' is an integer equal to 1 , 2, 3, or 4;

- B' is (CH ₂ ) _q - R'ii wherein q' is an integer equal to 0 or 1 and R'ii is selected from the group consisting of an amino group, a morpholine ring, a 4-methyl-piperazinyl group, a piperidinyl group, a methyloxy radical and a hydroxyl radical; - the arrow represents the attachment point of said solubilizing group to the carbon atom designated for V ₂ , V ₃ and V ₄ in the corresponding group of formula (Vl') as above-defined.

According to a preferred embodiment of the invention, R' ₃ represents one of the following groups:

According to a preferred embodiment of the invention, the solubilizing groups represent one of the following groups (S'-1 ') to (S'-8):

(S'-6) (S'-7) (S'-8)

In the groups -NH-A'-R' ₅ and in the groups of formulas (ll'-a) to (ll'-c) mentioned for R' ₄ , A' preferably represents -CH ₂ -, -CH(CH ₃ )-, -C(CHs) ₂ -, -CH ₂ -CH ₂ -, -C(CHs) ₂ -CH ₂ - or -CH ₂ -CH ₂ -O-.

In the groups of formula (ll'-a) to (ll'-c), R' ₅ preferably represents one of the following groups of formulas (IV-1 ) to (IV-34):

(IV-1 ) (IV-2) (IV-3) (IV-4)

(IV-9) (IV-10) (IV-11 )

(IV-15)

(IV-16) (IV-17)

(IV-19)

(IV-20)

(IV-21) (IV-22)

(IV-23) (IV-24)

(IV-25) (IV-26) (IV-27)

(lV-32) (IV-33) and

(IV-34)

As particular compounds of formula (I ¹ ), one can cite: (6RJ2aR)-2-(1-Benzyl-piperidin-4-yl)-6-phenyl-2,3,6J,12,12a- hexahydro- pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(6-chloro-pyrid in-3-yl)-2 _J 3 _I 6 _I 7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6S,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(6-chloro-pyrid in-3-yl)-2 _I 3,6,7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; - (eR.^aR^-CI-Benzyl-piperidin^-ylJ-e^θ-chloro-pyridin-S-yO^. S^,?,^, 12a-hexahydro-pyrazino[1 ',2': 1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[3-(benzyl-methyl-amino)-propyl]-2,3,6, 7,12,12a-hexahydro-pyrazino[1 \2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6S.12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[3-(benzyl-methyl-amino)-propyl]-2,3,6, 7,12, 12a-hexahydro-pyrazino[1\2^1 ,6]pyrido[3,4-b]indo!e-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[2-(benzyl-methyl-amino)-ethyl]-2,3,6,7, 12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3 _> 4-ib]indole-1 ,4-dione;

- (6S,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[2-(benzyl-methyl-amino)-ethyl]-2,3,6,7, 12, 12a-hexahydro-pyrazino[1\2':1 ,6]pyrido[3,4-ib]indole-1 ,4-dione;

- (6R _> 12aR>-6-Benzo[1 ,3]dioxol-5-yl-2-(1-benzyl-pipericlin-4-yl>-2 _I 3 _I 6,7,12,12a- hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-(1-benzyt-piperidin-4-yl)-9-fluoro-2,3,6, 7, 12, 12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-b3indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-(1-benzyl-piperidin-4-yl)-9-fluoro- 2,3,6,7,12, 12a-hexahydro-pyrazino[1\2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-benzo[1 ,3]dioxol-5-ylmethyl- pyrrolidin-3-yl)-2, 3,6,7,12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido [3,4~<b]indole -1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(4-methoxy-benzyl)-pyrrolidin-3- ylJ^.S^J.^.^a-hexahydro-pyrazinoπ'^'il .δjpyridop^-^indole-i ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(4-dimethyl-amino-benzyl)- pyrrolidin-S-yl^.S.ej.^.^a-hexahydro-pyrazinoπ'^'il .eipyridofS^-^indole- 1 ,4-dione;

- 2-[(R)-3-((6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-1 ,4-dioxo-3,4,6,7,12,12a-hexa hydro-1H-pyrazino[1',2 ^I :1 ,6]pyrido[3,4-b]indol-2-yl)-pyrrolidin-1-yl]-ethyl}- carbamic acid terf-butyl ester;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[1-(1-methyl-1/-/-imidazol-2-ylmethyl)- piperidin-4-yl]-2,3,6,7,12,12a-hexahydro-pyrazino[r,2':1 ,6]pyrido[3,4-b]indoIe- 1 ,4-dione;

- (6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6-(4-difluoromethoxy-p henyl)-2,3,6 ₎ 7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6-(2,2-difluoro-benzo[ 1 ,3]dioxol-5-yl)-2, 3,6,7, 12, 12a-hexahydro-pyrazino[1\2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,2 _I 5]thiadiazol-5-yl-2-((R)-1-benzyl-pyrrolidin-3-yl)-2 _> 3, 6,7,12,12a-hexahydro-pyrazino[1 \2':1 ,6]pyrido[3,4-fo]indole-1 ,4-dione;

- (6S,12aR)-6-Benzo[1 ,2,5]thiadiazol-5-yl-2-((R)-1-benzyl-pyrrolidin-3-yl)-2,3, 6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; - (6R,12aR)-6-Benzyl-2-(1-benzyl-piperidin-4-yl)-2,3,6,7,12,12 a-hexahydro- pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6S,12aR)-6-Benzyl-2-(1-benzyl-piperidin-4-yl)-2,3,6,7,12 ,12a-hexahydro- pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzyl-2-[1-(1-methyl-1H-imidazol-2-ylmethyl)-pi peridin-4-yl]-2, 3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

- (6S,12aR)-6-Benzyl-2-[1-(1-methyl-1H-imidazol-2-ylmethyl)-pi peridin-4-yl]-2, 3,6,7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

(6R,12aR)-2-{(R)-1 -[4-(2-{2-[2-(2-Amino-ethoxy)-ethoxy]-ethoxy}-ethoxy)- benzyl]-pyrrolidin-3-yl}-6-benzo[1 ,3]dioxol-5-yl-2,3,6,7,12,12a-hexahydro- pyrazino[1 \2':1 ,6]pyrido[3,4-/?]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1 -[3-(2-dimethylamino-ethoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4- b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(2-dimethylamino-ethoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4- £>]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[3-(3-morpholin-4-yl-propoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4- jb]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-{4-[3-(4-methyl-piperazin-1-yl)- propoxy]-benzyl}-pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro- pyrazino[1 ',2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[3-(2-morpholin-4-yl-ethoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4- jb]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(2-morpholin-4-yl-ethoxy)- £>]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(3-morpholin-4-yl-propoxy)- /b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(3-piperidin-1-yl-propoxy)- benzyl]-pyrrolidin-3-yl}-2 _I 3 _I 6,7,12 ₎ 12a-hexahydro-pyrazino[r,2 ^l :1 _> 6]pyrido[3 _> 4- £>]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(3-dimethylamino-propoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6 _l 7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4- b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[3-(2-methoxy-ethoxy)-benzyl]- pyrrolidin-S-ylj^.S.ej^Z.^a-hexahydro-pyrazinotr^'ii ^jpyridop^- b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[4-(2-methoxy-ethoxy)-benzyl]- b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-{(R)-1-[3,4-bis-(2-methoxy-ethoxy)- benzyO-pyrrolidin-S-yl^.S.δJ.^.^a-hexahydro-pyrazinotr^'il. ejpyridofS^- £>]indole-1 ,4-dione;

- (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(2,2-difluoro-b enzo[1,3]dioxol-5- yl)-2,3,6,7,12,12a-hexahydro-pyrazino[r,2 ^l :1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[1-(1-methyl-1H-imidazol-2-ylmethyl)- piperidin-4-yl]-2 _I 3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indoIe-

1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((S)-1-benzyl-pyrrolidin-3-yl)- 2,3,6, 7,12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(3,4-dimetho xy-phenyl)- 2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

(6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-phenyl-2,3,6 ,7,12,12a- hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-benzyl-pyrrolidin-3-yl)-9-fluoro- 2 _l 3,6,7,12,12a-hexahydro-pyrazino[1 ^l _J 2 ^I :1 ,6]pyrido[3,4-jb]indo!e-1 ,4-dione;

(6R,12aR)-2-((R)-1 -Benzyl-pyrrolidin-3-yl)-6-(3,4-dihydroxy-phenyl)- 2,3,6,7,12,12a-hexahydro-pyrazino[r,2':1 _l 6]pyrido[3,4-jb]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(2-methyl-benzyl)-pyrrolidin-3-yl]- 2,3,6,7,12,12a-hexahydro-pyrazino[1',2 ^I :1 ,6]pyrido[3,4-)b]indole-1 _I 4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(2-chloro-benzyl)-pyrrolidin-3-yl]- 2,3,6,7,12,12a-hexahydro-pyrazino[1 \2':1 ,6]pyrido[3,4-<b]indole-1 ,4-dione; yl]-2,3,6, 7, 12, 12a-hexahydro-pyrazino[1',2':1 ,6]pyrido[3,4-it>]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(2,4-difluoro-benzyl)-pyrrolidin-3- yl]-2,3,6, 7, 12, ^a-hexahydro-pyrazinotr^M .ejpyridotS^-^indole-i ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-benzyl-pyrrolidin-3-yl)-10-fluoro- 2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; - (6R,12aR)-2-((R)-1 -Benzyl-pyrrolidrn-3-yl)-6-(3-hydroxy-4-methoxy-phenyl)- ,4-dione;

- (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6-(4-hydroxy-3-me thoxy-phenyI)- 2,3,6,7,12, 12a-hexahydro-pyrazino[r,2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,2,5]thiadiazol-5-yl-2-{(R)-1-[3-(2-methoxy-ethoxy)- benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4- b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,2,5]thiadiazol-5-yl-2-{(R)-1-[3,4-bis-(2-methoxy- ethoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1 ,6]pyrido[3,4-j6]indole-1 ,4-dione; - (6R,12aR)-6-Benzo[1 ,2,5]thiadiazol-5-yl-2-{(R)-1-[4-(2-methoxy-ethoxy)- benzyI]-pyrrolidin-3-yl}-2,3,6,7,12,12a-hexahydro-pyrazino[r ,2':1,6]pyrido[3,4- b]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(1-methyl-1-phenyl-ethyl)- pyrrolidin-3-yl]-2,3,6,7,12,12a-hexahydro-pyrazino[r,2':1 ,6]pyrido[3,4-b]indole- 1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(1-phenyl-ethyl)-pyrrolidin-3-yl]- 2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-jb]indole-1 ,4-dione;

(6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1-phenethyl-pyrrolidin-3-yl)- 2,3,6, 7,12, 12a-hexahydro-pyrazino[1\2^1 ,6]pyrido[3,4-6]indole-1 ,4-dione;

- (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(2-phenoxy-ethyl)-pyrrolidin-3-yl]- 2,3,6,7,12,12a-hexahydro-pyrazino[1 ',2 ^l :1 ,6]pyrido[3,4-ib]indole-1 ,4-dione; - (6R, 12aR)-6-(3a,7a-Dihydro-benzo[1 ,3]dioxol-5-yl)-2-[(R)-1 -(1 ,1 -dimethyl-2- phenyl-ethyl)-pyrrolidin-3-yl]-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1 ,6]pyrido[3,4-/?]indole-1 ,4-dione;

(6R,12aR)-6-(3a _> 7a-Dihydro-benzo[1 ,3]dioxol-5-yl)-2-{1-[3-(2-methoxy- ethoxy)-benzyl]-azetidin-3-yl}-2,3,6,7,12,12a-hexahydro- pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione;

- (6R,12aR)-2-{1-[3,4-Bis-(2-methoxy-ethoxy)-benzyl]-azetidin- 3-yl}-6-(3a,7a- dihydro-benzoti .SJdioxol-δ-yO^^.θJ.^.^a-hexahydro- pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione; and pharmaceutically acceptable salts thereof. The chiral tetra-hydro β-carboline derivatives of formula (I) according to the invention can be prepared according to the general synthesis scheme A below:

SCHEME A in which Ri to R ₅ , A and k have the same significations than those previously indicated for compounds of formula (I) and Protec represents a protecting group. Protecting groups are well known from the one skilled in the art and are described for example in "Protective groups in organic synthesis", T.W. GREENE et a/., 2 ^nd edition, Wiley Interscience, 1991. As examples of these protecting groups, one can mention benzyl; benzyloxycarbonyle (Z); trifluoroacetyle (TFA); ferf-butyloxycarbonyle (Boc); trimethylsilylethoxycarbonyle (Teoc) and fluorenylmethyloxycarbonyle (Fmoc) groups.

According to this general process, and in a first step, tryptophan (a) is heated with an aldehyde derivative (b) in the presence of an acid according to the Pictet-Spengler reaction (Pictet, A.; Spengler, T., Ber., 1911 , 44, 2030) to give the corresponding compound (c). In a second step compound (c) is acylated with chloroacetyl chloride to lead to compound (d) which, in a third step, is cyclizated with an amine derivative bearing the R ₄

radical to lead to the expected compound of formula (I). These reactions are classically used in organic chemistry and are well known from the one skilled in the art.

Another subject matter of the invention is also the chiral tetra- hydro β-carboline derivatives of formula (T) for a use as a drug, in particular as a drug for the prevention and/or the treatment of parasitic diseases involving parasites having a phosphodiesterase (PDE) activity. According to a very prefered embodiment of the invention the drug is for the prevention and/or the treatment of malaria. Another subject matter of the invention is also a pharmaceutical composition comprising, as an active principle, at least one compounds of formula (I ¹ ), and at least one pharmaceutically acceptable excipient.

Finally, a subject matter of the invention is also a pharmaceutical composition comprising as an active principle, at least one compounds of formula (I ¹ ), and one or more additional antiparasitic drugs.

Besides the arrangements above, the invention also comprises other arrangements which will emerge from the following description, which refers to examples of preparation of β-carboline derivatives of formula (I) according to the invention, to in vitro demonstration of the antiparasitic activity of compounds of formula (I) and to in vivo demonstration of the antiparasitic activity of compounds of formula (I).

It should be clearly understood, however, that these examples are given only by way of illustration of the subject of the invention, of which they in no way constitute a limitation.

GENERAL SYNTHESIS PROTOCOLS USED IN THE FOLLOWING

EXAMPLES

I) General Protocol 1

General protocol 1 corresponds to a Pictet-Spengler reaction. D-tryptophan-methylester hydrochloride (D-tryptophan-OMe.HCI) and the aldehyde compound (1 eq.) were dissolved in dry dichloromethane (CH ₂ CI ₂ )

(0.05 M) containing activated molecular sieve (4?). The solution was cooled to

O ⁰ C. 3 eq. of trifluoroacetic acid in CH ₂ CI ₂ were added dropwise and the mixture was allowed to warm to room temperature and stirred for 24 h. The reaction mixture was filtered and partially evaporated. The crude product was diluted with ethyl acetate and washed with aqueous NaHCO ₃ and water. The organic layer was dried over magnesium sulphate (MgSO ₄ ) and evaporated to dryness. The diastereoisomers were separated by column chromatography. II) General Protocol 2: For acylation of β-carbolines intermediates β-carboline intermediates (1 eq.) were suspended in chloroform (0.2M) with 2.4 eq. of triethylamine (TEA). The reaction mixture was cooled at -10 ⁰ C and 2.4 eq. of chloroacetyl chloride in chloroform (0.5M) were added dropwise. The reaction mixture was stirred at -10 ⁰ C until completion and quenched with water. Organic layer was washed with aqueous solution of sodium hydrogenocarbonate (NaHCOs), and brine, dried over MgSO ₄ and evaporated to dryness. III) General Protocol 3: For cyclisation of chloro-acetyl intermediates with amines

The chloro-acetyl intermediates (1 eq.) and amine (2 eq.) are refluxed in methanol or ethanol (0.075 M) for 12-24 h. The methanol was removed by evaporation. The residue was dissolved in ethyl acetate, washed with saturated NaHCO ₃ aqueous solution, dried over MgSO ₄ , evaporated to dryness and purified by flash chromatography in a dichloromethane/methanol mixture (CH ₂ CI ₂ /MeOH). IV) General Protocol 4: For reductive amination: Procedure 4

The intermediates 23 or 24 as described hereinafter (1 eq.) and the aldehyde (1.5 eq.) were dissolved in a mixture of dichloromethane with acetic acid (CH ₂ CI ₂ /acetic acid) 10/1 (0.1 M). 2 eq. of (polystyrylmethyl) trimethylammonium cyanoborohydride resin were suspended and the reaction mixture was stirred at room temperature for 12-24 hours. The precipitate was dissolved with methanol, the resin was filtered, the solution evaporated to dryness. The reaction mixture was purified by thin layer chromatography (TLC) with a solvent system CH ₂ CI ₂ /MeOH.

V) General Protocol 5: Synthesis of methylic ester of tryptophan derivatives

Thionyl chloride (3 eq.) was added dropwise to a suspension of tryptophan derivative (1 eq.) in anhydrous methanol at 0 ⁰ C. The mixture was warmed slowly to room temperature and stirred for 24 h. The thionyl chloride treatment was repeated an additional time as described above. The methanol and the thionyl chloride excess were removed under reduced pressure.

VI) General Protocol 6: For cyclisation of acylated intermediates with amine in microwaves

Acylated intermediates were dissolved in ethanol (0.04 M) with 2.5 eq. of amine. The reaction mixture was heated by microwaves 100 W for 2-15 minutes. The reaction mixture was totally evaporated and purified by thick layer chromatography. EXAMPLE 1 : Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-((R)-1- benzyl-pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro-pyrazino[1' ,2':1,6]pyrido [3,4-Jb] indole-1,4-dione (Compound 6).

1) First step: Synthesis of the intermediate (1 R,3R)-1-Benzo[1 ,3]dioxol-5-yl- 2,3,4,9-tetrahydro-i H-β-carboline-3-carboxylic acid methyl ester (Compound 2)

2 was prepared according to general protocol 1 starting from D-tryptophan OMe. HCI (3 g) and obtained as 950 mg of a white powder (23%).

LC : t _R =3.7 min; MS (ESI+): m/z=351 (M+H) ⁺ ;

¹ H NMR (CDCI ₃ ): d 7.45 (d, J=6.9 Hz, 1 H) ₁ 7.39 (s, 1 H), 7.16

(m, 1 H) ₁ 7.10-7.01 (m, 2H), 6.81 (dd, J=8.0 Hz and J=M Hz, 1 H), 6.75 (d,

J=M Hz, 1 H), 6.73 (d, J=8.0 Hz, 1 H), 5.87 (s, 2H), 5.09 (s, 1 H), 3.88 (dd,

J=11.1 Hz and J=4.3 Hz, 1 H), 3.75 (s, 3H), 3.14 (ddd, J=15.0 Hz, J=4.3 Hz and J=1.9 Hz, 1 H), 2.91 (ddd, J= 15.0 Hz, J= 11.0 Hz and J=2.5 Hz, 1 H);

¹³ C NMR (CDCI ₃ ), d 173.58, 148.58, 148.23, 136.56, 135.08, 127.52, 122.39, 120.04, 118.64, 111.36, 109.26, 109.20, 108.72, 101.64, 58.8, 57.27, 52.68, 26.07,

Mp=160-161 °C. 2) Second step: Synthesis of the intermediate (1 R,3R)-1-Benzo[1 ,3]dioxol-5- yl-2-(2-chloro-acetyl)-2,3,4,9-tetrahydro-1/-/-β-carboline- 3-carboxylic acid methyl ester (Compound 5)

5 was prepared according to general protocol 2 starting from intermediate 2 of step 1 (2.0 g) and obtained after precipitation in diethyl ether as a white powder (2 g, 83%). LC: t _R =6.2 min; MS (ESI+): m/z = 427 (M+H) ⁺ ;

¹ H NMR (CDCI ₃ ): d 7.87 (s, 1 H), 7.61 (d, J=7.8 Hz, 1 H), 7.15- 7.32 (m, 3H), 6.84-6.90 (m, 2H), 6.65 (brs, 2H), 5.92 (s, 2H), 4.95 (brs, 1 H), 4.20-4.40 (m, 2H), 3.70 (d, J=16.2 Hz, 1 H), 3.22 (m, 4H).

3) Third Step: Synthesis of Compound 6

Compound 6 was prepared according to general protocol 3 starting from intermediate 5 as obtained here-above in step 2 (1.55 g) and (R)-

(-)-1-Benzyl-3-aminopyrrolidine (2 eq.). Compound 6 was obtained after purification by flash chromatography (CH ₂ CI ₂ /MeOH 99/1 : v/v) as a white powder (936 mg, 48%).

LC: t _R =4.9 min;

MS (ESI+): m/z = 535 (IvHH) ⁺ ;

¹ H NMR (CDCI ₃ ) : d 8.25 (s, 1 H), 7.60 (d, 1 H, J=6.6 Hz, 1 H), 7.14-7.37 (m, 8H), 6.84 (dd, J=8.1 Hz and J=A .5 Hz ₁ 1 H), 6.75 (d, J=1.5 Hz ₁ 1 H), 6.68 (d, J=8.1 Hz, 1 H), 6.24 (s, 1 H), 5.86 (d, J=3.6 Hz, 2H), 5.12 (brs, 1 H), 4.41 (brm, 1 H), 4.22 (dd, J=11.4 Hz and J=4.8 Hz, 1 H), 3.99 (d, J=Ml Hz, 1 H), 3.78 (brs, 2H), 3.70 (dd, J=15.9 Hz and J=4.8 Hz, 1 H), 3.22 (dd, J=A 6.2 Hz and J=11.7 Hz, 1 H), 1.79-3.09 (m, 6H). EXAMPLE 2: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-(1-benzyl- piperidin-4-yl)-2,3,6J,12,12a-hexahydro-pyrazino[1\2':1,6]py rido[3,4- jb]indole-1,4-dione (Compound 10)

Compound 10 was prepared according to general protocol 3 starting from intermediate 5 as above-obtained at step 2 of Example 1 (100 mg) and 4-amino-N-benzylpiperidine (2 eq.) in ethanol (EtOH). Compound 10 was obtained after purification by flash chromatography (CH ₂ CI ₂ /MeOH) as a white powder (76 mg, 59%).

LC: t _R =4.7 min;

MS (ESI+): m/z = 549 (M+H) ⁺ ;

¹ H NMR (MeOD) : d 7.54 (d, J=7.2 Hz, 1 H), 7.27-7.34 (m, 6H), 7.06 (m, 2H), 6.79 (dd, J=8.1 Hz and J=1.8 Hz, 1H), 6.75 (d, J=1.8 Hz, 1H), 6.67 (d, J=8.1 Hz, 1 H), 6.21 (s, 1 H), 5.82 (d, J=3.6 z, 2H), 4.37 (m, 2H), 4.06 (d, J=17.1 Hz, 1 H), 3.96 (d, J=17.1 Hz ₁ 1 H), 3.63 (dd, J=15.9 Hz and J=4.8 Hz, 1 H) ₁ 3.53 (s, 2H), 3.13 (dd, J=15.0 Hz and J=11.4 Hz, 1 H), 2.97 (m, 2H), 2.09 (m, 2H), 1.68-1.88 (m, 3H), 1.54 (m, 1 H).

EXAMPLE 3: Synthesis of (6S,12aR)-6-Benzo[1,3]dioxol-5-yl-2-((R)-1- benzyl-pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro-pyrazino[1' ,2':1,6]pyrido [3,4-b]indole-1,4-dione (Compound 14)

1) First Step: Synthesis of the intermediate (1S,3R)-1-Benzo[1 ,3]dioxol-5-yl- 2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (Compound 3)

3 was prepared according to general protocol 1 starting from D-tryptophan OMe. HCI (3 g) and obtained as 870 mg of a white powder (21%). LC: t _R =3.9 min;

MS (ESI+): m/z=351 (M+H) ⁺ ;

¹ H NMR (CDCI ₃ ), d 7.52 (s, 1 H), 7.47 (d, J=6.8 Hz, 1 H), 7.17

(m, 1H), 7.11-7.02 (m, 2H), 6.67 (s, 3H), 5.85 (s, 2H), 5.25 (s, 1H), 3.90 (t,

J=6.2 Hz, 1 H), 3.63 (s, 3H), 3.18 (dd, J=15.5 Hz and J=5.5 Hz, 1 H), 3.04 (dd,

J=15.3 Hz and J=6.6 Hz, 1 H); ¹³ C NMR (CDCI ₃ ), d 174.4, 148.46, 147.90, 136.60, 136.15,

133.52, 127.34, 122.43, 122.25, 119.93, 118.68, 111.37, 109.18, 108.56,

101.59, 55.06, 53.88, 52.91 , 52.58, 24.93; Mp=187-189°C

2) Second Step: Synthesis of the intermediate (1S,3R)-1-Benzo[1 ,3]dioxol-5- yl-2-(2-chloro-acetyl)-2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (Compound 4)

4 was prepared according to general protocol 2 starting from intermediate 3 as obtained here-above at step 1 (996 mg, 1 eq.). Compound 4 was obtained as a yellow powder (1.06 g, 96%), which was directly engaged in the cyclisation step.

LC: t _R =6.3 min;

MS (ESI+): m/z=427 (M+H) ⁺ . 3) Third Step: Synthesis of Compound 14 Compound 14 was prepared according to general protocol 3 starting from intermediate 4 as here-above obtained at step 2 (1.55 g) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (2 eq.). Compound 14 was obtained after purification by flash chromatography (CH ₂ CI ₂ ZMeOH 99/1 : v/v) as a white powder (936 mg, 48%). LC: t _R =4.8 min;

MS (ESI+): m/z = 535 (M+H) ⁺ ;

¹ H NMR (CDCI ₃ ) : d 7,88 (s, 1H) ₁ 7.52 (d, J=7,6 Hz, 1 H), 7.32-

7.13 (m, 8H), 6.96 (s, 1 H), 6.81 (s, 1 H), 6.71 (s, 2H), 5.93 (s, 2H), 5.22 (brs, 1 H), 4.33 (dd, J=11.7 Hz and J=4.2 Hz ₁ 1 H), 4,29 (d, J=M.6 Hz ₁ 1 H), 4,12 (d, J=18,0 Hz, 1 H), 3.57 (s, 2H), 3.50 (dd, J= 15.1 Hz and J=3.8 Hz, 1 H), 2,93 (m, 2H), 2,73 (m, 1 H), 2,47 (dd, J=10,5 Hz and J=7,6 Hz, 1 H), 2,21 (m, 2H), 1.75 (m, 1 H).

EXAMPLE 4: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[2-(benzyl- methyl-amino)-ethyl]-2,3,6,7,12 _J 12a-hexahydro-pyrazino[1',2':1,6]pyrido [3,4-Jb]indole-1,4-dione (Compound 15)

1) First Step: Synthesis of a reactive amine: N1-Benzyl-N1-methyl-ethane- 1 ,2-diamine, 2HCI (Compound 20)

a) Synthesis of [2-(Benzyl-methyl-amino)-ethyI]-carbamic acid ferf-butyl ester (Compound 19)

N-Methyl-benzylamine (114 μL, 1 eq.), 2-(Boc- amino)ethylbromide (200 mg, 1 eq.), potassium iodide (74 mg, 0.5 eq.) and cesium carbonate (580 mg, 2 eq.) were dissolved in acetone and stirred at room temperature for 70 h. The reaction mixture was diluted with ethyl acetate, washed with water, dried over MgSO ₄ , and evaporated to dryness to give 160 mg of yellow oil (68 %).

LC: t _R =3.6 min; .

MS (ESI+): m/z = 265 [M+H] ⁺ ;

¹ H NMR (CD ₂ CI ₂ ) : d 7,38-7,26 (m, 5H), 5,05 (m, 1 H), 3,53 (s, 2H), 3,23 (q, J=5,8 Hz, 2H), 2,51 (t, J=6,0 Hz, 2H), 2,21 (s, 3H), 1 ,47 (s, 9H). b) Synthesis of (Compound 20)

Intermediate 19 (160 mg, 600 μmol) was dissolved in dioxane (3 mL), 1.5 ml_ of HCI 4N in dioxane was added, The reaction mixture was stirred at room temperature for 2Oh. Dioxane was removed by evaporation to give 125 mg of translucent oil (88%) to lead to compound 20. LC: t _R =1.1 min;

MS (ESI+): m/z = 165 [M+H] ⁺ . 2) Second Step: Preparation of Compound 15

Compound 15 was prepared according to general protocol 3 starting from intermediate 5 as obtained here-above at step 2 of Example 1 (100 mg), Compound 20 as prepared above at Step 1) b) (2 eq.) and TEA (3 eq.) in EtOH. Compound 15 was obtained after purification by flash chromatography (CH ₂ CI ₂ /MeOH) as a yellow powder (50 mg, 41 %).

LC: t _R =4.6 min;

MS (ESI+): m/z = 523 (M+H) ⁺ ; ¹ H NMR (MeOD): d 7.54 (d, J=7.2 Hz, 1 H), 7.20-7.30 (m, 6H),

7.07 (m, 2H), 6.83 (m, 2H), 6.65 (d, J=7.8 Hz, 1H), 6.24 (s, 1 H), 5.81 (d, J=5.4 Hz, 2H), 4.37 (dd, J=11.4 Hz and J=3.6 Hz, 1 H), 4.20 (dd, J=MA Hz and J=1.5 Hz, 1 H), 4.01 (d, J=MA Hz, 1 H), 3.47-3.69 (m, 5H), 3.12 (dd, J=15.9 Hz and J= 11.4 Hz, 1 H), 2.62 (m, 2H), 2.23 (s, 3H).

EXAMPLE 5: Synthesis of (6S,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[2-(benzyl- methyl-amino)-ethyl]-2, 3,6,7, 12,12a-hexahydro-pyrazino[1',2":1,6]pyrido [3,4-Jb]indole-1,4-dione (Compound 16)

Compound 16 was prepared according to general protocol 3 starting from intermediate 4 as obtained here-above at step 2 of Example 3 (56 mg), Compound 20 as obtained above at Step 1 ) b) of Example 4 (4 eq.) and TEA (12 eq.) in MeOH (48 h). Compound 16 was obtained after purification by flash chromatography (CH ₂ CI ₂ ZMeOH) as a white powder (30 mg. 41 %).

LC: t _R =4.9 min; MS (ESI-): m/z = 523 (M-H);

¹ H NMR (MeOD) : d 7.51 (d, J=7.8 Hz, 1 H) ₁ 7.33 (d, J=7.8 Hz ₁

1 H), 7.21-7.25 (m, 2H), 7.05-7.25 (m, 5H), 6.99 (s, 1 H), 6.85 (d, J=1.5 Hz, 1H), 6.80 (d, J=8.1 Hz, 1 H), 6.70 (dd, J=8.1 Hz and J=1.5 Hz,1 H), 5.94 (d,

J=6.6 Hz, 2H), 4.15-4.34 (m, 3H), 3.55 (m, 4H), 3.34 (m, 1 H), 2.90 (ddd,

J=15.3 Hz, J=11.7 Hz and J= 1.2 Hz, 1 H), 2.57 (m, 2H), 2.33 (s, 3H).

EXAMPLE 6: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[3-(benzyl- methyl-amino)-propyl]-2,3,6,7,12 ₎ 12a-hexahydro-pyrazino[1',2 ^t :1,6]pyrido [3,4-Jb]indole-1,4-dione (Compound 17)

1) First Step: Synthesis of a reactive amine: N1-Benzyl-N1-methyl-propane- 1 ,3-diamine, 2HCI (Compound 22)

a) Synthesis of [3-(Benzyl-methyl-amino)-propyl]-carbamic acid te/t-butyl ester (Compound 21)

N-Methyl-benzylamine (108 μL, 1 eq.), 2-(Boc- amino)propylbromide (200 mg, 1 eq.), potassium iodide (69 mg, 0.5 eq.) and cesium carbonate (547 mg, 2 eq.) were dissolved in acetone and stirred at room temperature for 70 h. The reaction mixture was diluted with ethyl acetate, washed 3 times with saturated NaHCO ₃ aqueous solution, dried over MgSO ₄ , and evaporated to dryness to give 210 mg of yellow oil (90 %). LC: t _R =3.4 min; MS (ESI+): m/z = 279 [M+H] ⁺ ;

¹ H NMR (CD ₂ CI ₂ ): d 7.34-7.25 (m, 5H), 5.62 (m, 1 H), 3.51 (s, 2H), 3.18 (q, J=6.1 Hz, 2H), 2.47 (t, J=6.3 Hz, 2H), 2.18 (s, 3H), 1.69 (qt, J=6.4 Hz, 2H), 1.47 (s, 9H). b) Synthesis of Compound 22

Intermediate 21 as obtained above at step a) (210 mg) was

dissolved in dioxane (3 ml_), 1.5 ml_ of HCI 4N in dioxane was added, The reaction mixture was stirred at room temperature for 2Oh. Dioxane was removed by evaporation to give 190 mg of translucent oil (90%).

LC: t _R =1.1 min; MS (ESI+): m/z = 179 [M+H] ⁺ .

2) Second Step: Synthesis of Compound 17

Compound 17 was prepared according to general protocol 3 starting from intermediate 5 as obtained here-above at step 2 of Example 1

(100 mg), Compound 22 as above obtained at Step 1 ) b) (2 eq.) and TEA (3 eq.) in EtOH (5 days). Compound 17 was obtained after purification by flash chromatography (CH ₂ CI ₂ ZMeOH) as a white powder (30 mg, 24%).

LC: t _R =4.7 min;

MS (ESI+): m/z = 537 (M+H) ⁺ ;

¹ H NMR (MeOD) : d 7.54 (d, J=6.9 Hz, 1 H), 7.21-7.33 (m, 6H), 7.07 (m, 2H), 6.81 (m, 2H), 6.68 (d, J=8.1 Hz, 1 H), 6.23 (s, 1 H), 5.83 (s, 2H), 4.36 (dd, J=11.7 Hz and J=3.9 Hz, 1 H), 4.22 (dd, J=17.1 Hz and J=1.2 Hz, 1 H), 3.99 (d, J=17.1 Hz, 1 H), 3.62 (dd, J=15.6 Hz and J=4.8 Hz, 1 H), 3.52 (m, 4H), 3.12 (dd, J=14.7 Hz and J=11.4 Hz, 1 H), 2.44 (td, J=6.9 Hz and J=3.0 Hz, 2H), 2.20 (s, 3H), 1.85 (qt, J=7.5 Hz, 2H). EXAMPLE 7: Synthesis of (6S,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[3-(benzyl- methyl-amino)-propyl]-2,3 _J 6,7,12,12a-hexahydro-pyrazino[1',2':1,6]pyrido [3,4-b]indole-1,4-dione (Compound 18)

Compound 18 was prepared according to general protocol 3 starting from intermediate 4 as obtained her-above at step 2 of Example 3 (86 mg), Compound 22 as above obtained at step 1) b) of Example 6 (4 eq.) and

TEA (12 eq.) in MeOH (48 h). Compound 18 was obtained after purification by flash chromatography (CH ₂ CI ₂ /MeOH) as a yellow powder (70 mg, 65%).

LC: t _R =4.7 min;

MS (ESI-): m/z = 537 (M-H);

¹ H NMR (MeOD) : d 7.51 (d, J=7.5 Hz, 1 H), 7.24-7.34 (m, 5H), 7.05-7.18 (m, 3H), 6.95 (s, 1 H), 6.80 (d, J=1.5 Hz, 1 H), 6.76 (d, J=8.1 Hz, 1 H), 6.66 (dd, J=8.1 Hz and J=I .5 Hz ₁ IH), 5.93 (d, J=2.4 Hz, 2H), 4.19-4.32 (m, 2H), 4.08 (d, J=MA Hz, 1 H), 3.48-3.60 (m, 4H), 3.36 (m, 1 H), 2.86 (ddd, J=15.0 Hz, J=11.7 Hz and J=1.2 Hz, 1 H), 2.41 (m, 2H), 2.24 (s, 3H), 1.84 (qt, J=7.2 Hz, 2H).

EXAMPLE 8: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1-(4- dimethylamino-benzyl)-pyrrolidin-3-yl]-2, 3,6,7, 12,12a-hexahydro- pyrazinoII'^'ri.θlpyridoIS^-Jblindole-i^-dione (Compound 26)

1) First Step: Synthesis of the intermediate (6R,12aR)-6-Benzo[1,3]dioxol-5- yl-2-(R)-pyrrolidin-3-yl-2,3 _J 6,7,12 _I 12a-hexahydro-pyrazino[1' _I 2 ^l :1 ,6]pyrido[3,4- fa]indole-1 ,4-dione (Compound 23)

Compound 6 as above obtained at the end of Example 1 (720

mg, 1 eq.) was dissolved in ethyl acetate (0.07 M), and palladium dihydroxide (Pd(OH) ₂ ) on carbon (1 eq. w/w) was suspended. The reaction mixture was stirred at room temperature under H ₂ atmosphere for 24-48 h. The reaction mixture was filtered on celite, evaporated to dryness and precipitated in diethyl ether to obtain compound 23 as a white powder (412 mg, 69%).

LC: t _R =4.0 min;

MS (ESI+): m/z = 445 (M+H) ⁺ ;

¹ H NMR (DMSO-c/e) : d 11.12 (s, 1 H), 8.31 (s, 1 H), 7.54 (d, J=7.8 Hz, 1 H), 7.30 (d, J=7.8 Hz, 1 H), 7.08-6.97 (m, 2H), 6.83-6.72 (m, 3H), 6.17 (S ₁ 1 H), 5.92 (s, 1 H), 4.92 (m, 1 H), 4.43 (dd, J=11.1 Hz and 4.8 Hz, 1 H), 4.17 (d, J=16.6 Hz, 1 H), 4.00 (d, J=16.8 Hz, 1 H), 3.47 (dd, J=15.8 Hz and J=4.9 Hz, 1 H), 3.14-2.88 (m, 5H), 2.08-2.03 (m, 1 H), 1.75-1.66 (m, 1 H). 2) Second Step: Synthesis of Compoud 26

26 was prepared according to general protocol 4 starting from intermediate 23 as above obtained at step 1 (45 mg) and p- dimethylaminobenzaldehyde (1.5 eq.). Compound 26 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (16 mg, 27%).

LC: t _R =5.14 min;

MS (ESI+): m/z = 578 (M+H) ⁺ ; ¹ H NMR (MeOD) : d 7.52 (d, J=6.9 Hz, 1H), 7.28 (d, J=7.5 Hz,

1H), 7.20 (d, J=8.7 Hz, 1H), 7.06 (m, 2H), 6.66-6.79 (m, 5H), 6.24 (s, 1H), 5.85 (s, 2H), 5.04 (brs, 1H), 4.34 (dd, J=11.7 Hz and J=5.4 Hz, 1H), 4.31 (d, J=17.1 Hz, 1H), 4.09 (dd, J=17.1 Hz and J=1.2 Hz, 1H), 3.66 (d, J=12.3 Hz, 1H), 3.58 (dd, J=15.6 Hz and J=4.8 Hz, 1H), 3.46 (d, J=12.6 Hz, 1H), 3.12 (dd, J=15.9 Hz and J=11.7 Hz, 1H), 2.97 (m, 1H), 2.90 (s, 6H), 2.75 (dd, J=10.8 Hz and J= 4.2 Hz, 1H), 2.54 (dd, J=10.5 Hz and J=7.8 Hz, 1H) ₁ 2.25-2.38 (m, 2H), 1.19-1.72 (m, 1H).

EXAMPLE 9: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-((R)-1- pyridin-2-ylmethyl-pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro -pyrazino [1',2':1,6]pyrido[3,4-Jb]indole-1,4-dione (Compound 27)

27 was prepared according to general protocol 4 starting from intermediate 23 as above obtained at step 1 of Example 8 (45 mg) and pyridine-2-carboxaldehyde (1.5 eq.). Compound 27 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (27 mg, 50%). LC: t _R =4.58 min; MS (ESI+): m/z = 536 (IvRH) ⁺ ;

¹ H NMR (MeOD) : d 8.53 (d, J= 4.8 Hz, 1 H), 7.86 (td, J=7.8 Hz and J=1.8 Hz, 1 H), 7.55 (m, 2H), 7.36 (dd, J=6.6 Hz and J=5.1 Hz, 1 H), 7.29 (d, J=7.5 Hz, 1 H), 7.06 (m, 2H), 6.78-6.81 (m, 2H), 6.68 (d, J=8.7 Hz, 1 H), 6.25 (s, 1 H), 5.85 (s, 2H), 5.01 (brs, 1 H) ₁ 4.39 (dd, J=11.7 Hz and J=5.1 Hz, 1 H), 4.3 (d, J=MA Hz, 1 H), 4.19 (d, J= 17.7 Hz, 1 H), 3.92-4.03 (m, 2H), 3.61 (dd, J=15.6 Hz and J=4.8 Hz, 1 H), 3.07-3.25 (m, 3H), 2.78-2.84 (m, 1 H), 2.54-2.62 (m, 1 H), 2.32-2.43 (m, 1 H), 1.89-1.84 (m, 1 H).

EXAMPLE 10: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1-(1- methyl-1W-imidazol-2-ylmethyl)-pyrrolidin-3-yl]-2,3,6,7,12,1 2a-hexahydro- pyrazino[1\2':1,6]pyrido[3,4-jb]indole-1,4-dione (Compound 28)

28 was prepared according to general protocol 4 starting from intermediate 23 as above obtained at step 1 of Example 8 (45 mg) and 1- methyl-2-imidazolecarboxaldehyde (1.5 eq.). Compound 28 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (26 mg, 48%). LC: t _R =4.2 min; MS (ESI+): m/z = 539 (M+H) ⁺ ;

¹ H NMR (MeOD) : d 7.53 (d, J=6.9 Hz ₁ 1H), 7.29 (d, J=7.5 Hz, 1 H), 7.01-7.12 (m, 3H), 6.90 (d, J=1.2 Hz, 1 H), 6.75-6.79 (m, 2H), 6.67-6.69 (m, 1 H), 6.25 (s, 1H), 5.85 (s, 2H), 5.15 (brs, 1H), 4.37 (dd, J^l 1.4 Hz and J=4.5 Hz, 1 H), 4.26 (d, J=MA Hz, 1 H), 4.10 (d, J=MA Hz, 1 H), 3.78 (s, 3H), 3.73 (d, J=2.4 Hz, 2H), 3.59 (dd, J=15.6 Hz and J=5.1 Hz, 1H), 3.12 (dd, J=15.6 Hz and J=11.7 Hz, 1 H), 2.93 (m, 1 H), 2.80 (dd, J=10.8 Hz and J=2.1 Hz, 1H), 2.50 (dd, J=10.5 Hz and J=7.5 Hz, 1 H), 2.28 (m, 2H), 1.62-1.67 (m, 1 H).

EXAMPLE 11 : Synthesis of (6S,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1-(1- methyl-1H-imidazoI-2-ylmethyl)-pyrrolidin-3-yl]-2,3,6,7,12,1 2a-hexahydro- pyrazino[1\2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 29)

1) First Step: Synthesis of the intermediate (6S,12aR)-6-Benzo[1 ,3]dioxol-5- b]indole-1 ,4-dione (Compound 24)

Compound 14 as obtained above at the end of Example 3 (235 mg) was dissolved in ethyl acetate (0.07 M), and Pd(OH) ₂ on carbon (1 eq. w/w) was suspended. The reaction mixture was stirred at room temperature under H ₂ atmosphere for 24-48 h. The reaction mixture was filtered on celite, evaporated to dryness and precipitated in diethyl ether to obtain compound 24 as a white powder. LC: t _R =4.0 min;

MS (ESI+): m/z = 445 (IvRH) ⁺ . 2) Second Step: Synthesis of compound 29

29 was prepared according to general protocol 4 starting from intermediate 24 as above obtained at step 1 (45 mg) and 1-methyl-2-

imidazolecarboxaldehydθ (1.5 eq.). Compound 29 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (15 mg, 28%).

LC: t _R =4.36 min;

MS (ESI+): m/z = 539 (M+H) ⁺ ; ¹ H NMR (DMSO-d ₆ ) : d 11 ,07 (s, 1 H), 7.50 (d, J=8,1 Hz ₁ 1 H),

7.31 (d, J=8,1 Hz ₁ 1 H), 7.08 (m, 3H), 6.87 (d, J=8,1 Hz, 1 H), 6.76 (m, 3H), 6.60 (d, J=8,1 Hz, 1 H), 6.01 (m, 2H), 4,98 (brs, 1 H), 4,27 (d, J=17,4 Hz, 1 H), 4.07 (m, 2H), 3.66 (m, 4H), 3.54 (d, J=13,5 Hz, 1 H), 3.21 (dd, J=15.3 Hz and J=4,2 Hz, 1 H), 2,97 (dd, J=14,4 Hz and J=12,9 Hz, 1 H), 2,78 (m, 2H), 2.38 (m, 1 H), 2.17-2,03 (m, 2H), 1 ,80 (m, 1 H).

EXAMPLE 12: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1-(4- methoxy-benzyl)-pyrrolidin-3-yl]-2,3,6,7,12,12a-hexahydro- pyrazino[1\2':1,6]pyrido[3,4-b]indole-1,4-dione (Compoud 31)

31 was prepared according to general protocol 4 starting from intermediate 23 as above obtained at step 1 of Example 8 (50 mg) and para- anisaldehyde (2.0 eq.). Compound 31 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (20 mg, 31 %). LC: t _R =5.0 min; MS (ESI+): m/z = 565 (M+H) ⁺ ;

¹ H NMR (MeOD) : d 7.52 (d, J=7.2 Hz, 1 H), 7.29 (d, J=8.7 Hz, 3H), 7.06 (m, 2H), 6.89 (d, J=8.7 Hz, 2H), 6.66-6.79 (m, 3H), 6.24 (s, 1 H), 5.85 (s, 2H), 5.01 (m, 1 H), 4.34 (dd, J=11.1 Hz and J=4.8 Hz, 1 H), 4.30 (d, J=17.1 Hz, 1 H), 4.10 (d, J=17.1 Hz, 1 H), 3.77 (s, 3H), 3.73 (d, J=12.6 Hz, 1 H), 3.59 (dd, J=15.9 Hz and J=4.8 Hz, 1 H), 3.56 (d, J=12.6 Hz, 1 H), 3.12 (dd, J=15.6 Hz and J=11.7 Hz, 1 H), 3.03 (td, J=8.4 Hz and J=2.7 Hz, 1 H), 2.83

(dd, J=11.1 Hz and J=3.3 Hz, 1 H) ₁ 2.59 (dd, J=10.8 Hz and J=7.8 Hz, 1 H),

2.24-2.45 (m, 2H), 1.68-1.80 (m, 1 H).

EXAMPLE 13: Synthesis of (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-((R)-1- benzo[1,3]dioxol-5-ylmethyl-pyrrolidin-3-yl)-2,3,6,7,12,12a- hexahydro- pyrazinoJI'^'il.βlpyridotS^-Jblindole-i ^-dione (Compound 33)

33 was prepared according to general protocol 4 starting from intermediate 23 as above obtained at step 1 of Example 8 (45 mg) and Piperonal (1.2 eq.). Compound 33 was obtained after purification by TLC (CH ₂ CI ₂ /Me0H) as a white powder (18 mg, 31 %).

LC: t _R =4.8 min;

MS (ESI+): m/z = 579 (M+H) ⁺ ;

¹ H NMR (MeOD) : d 5.53 (d, J=6.9 Hz, 1H), 7.29 (d, J=7.5 Hz, 1H), 7.06 (m, 2H), 6.90 (d, J=1.2 Hz, 1H), 6.74-6.84 (m, 4H), 6.68 (m, 1H), 6.25 (S ₁ 1H), 5.92 (s, 2H), 5.85 (s, 2H), 5.05 (m, 1H), 4.37 (dd, J=11.7 Hz and J=4.2 Hz, 1H), 4.31 (d, J=16.8 Hz, 1H), 4.14 (d, J=15.9 Hz, 1H), 3.69 (d, J=12.6 Hz, 1H) ₁ 3.51-3.66 (m, 3H) ₁ 3.00-3.17 (m, 2H), 2.83 (dd, J=10.8 Hz and J=3.3 Hz, 1H) ₁ 2.58 (dd, J=10.8 Hz and J=7.8 Hz, 1H) ₁ 2.27-2.40 (m, 2H) ₁ 1.70-1.76 (m, 1H).

EXAMPLE 15: Synthesis of {2-[(R)-3-((6R,12aR)-6-Benzo[1,3]dioxol-5-yl- i^-dioxo-Sλej.^jaa-hexahydro-IW-pyrazinoII'^M^lpyridoIS^-ib ] indol-2-yl)-pyrrolidin-1-yl]-ethyl}-carbamic acid tert-butyl ester Compound 35)

Compound 23 as above obtained at step 1 of Example 8 (50 mg), 2-(Boc-amino)ethyl bromide (25 mg, 1 eq.) and triethylamine (1.1 eq.) were dissolved in dioxane (1 ml) and stirred at room temperature. 46 hours later, 0.5 eq. of 2-(Booamino)ethyl bromide, 0.5 eq. of TEA and 0.5 ml of dioxane were added. 96 hours later, the reaction mixture was heated to 70 ⁰ C for 20 hours. The dioxane was removed by evaporation; the residue was dissolved in ethyl acetate, washed with saturated NaHCOs aqueous solution, dried over MgSO ₄ and evaporated to dryness. Compound 35 was purified by TLC (CH ₂ CI ₂ /MeOH) to give 29 mg of a white powder (44%). LC: t _R = 4.94 min;

MS (ESI+): m/z = 588 (IvHH) ⁺ ;

¹ H NMR (MeOD) : d 7.53 (d, J=7.2 Hz, 1 H), 7.29 (d, J=7.2 Hz, 1 H), 7.07 (m, 2H,), 6.68-6.80 (m, 3H), 6.27 (s, 1 H), 5.86 (s, 2H), 5.10 (m, 1 H), 4.41 (dd, J=11.4 Hz and J=4.5 Hz ₁ 1 H), 4.31 (d, J=MA Hz, 1 H), 4.18 (d, J=MA Hz, 1 H), 3.61 (dd, J=15.6 Hz and J=4.8 Hz, 1 H), 2.87-3.23 (m, 5H), 2.50-2.63 (m, 3H), 2.23-2.32 (m, 2H), 1.65-1.71 (m, 1 H), 1.44 (s, 9H).

EXAMPLE 16: Synthesis of (6R,12aR)-6-Benzo[1,2,5]thiadiazol-5-yl-2- ((RJ-i-benzyl-pyrrolidin-S-yO-a.S _j θJj^.iaa-hexahydro-pyrazinoli'^'rijθ] pyrido[3,4-Jb]indole-1,4-dione (Compound 70c)

1) First Step: Synthesis of (1 R,3R)-1-Benzo[1 ,2,5]thiadiazol-5-yI-3- methoxycarbonyl-2,3,4,9-tetrahydro-1 H-β-carbolin-2-ium trifluoroacetate

(Compound 64c)

64c was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (1241 mg) and 2,1 ,3-benzothiadiazole-5- carbaldehyde (800 mg). The reaction mixture was stirred overnight. After filtration and concentration, the cis isomer was precipitated in EtOAc to provide the compound 64c as a yellow solid (1180 mg, 66%).

LC: t _R = 3.91 min; MS (ESI+): m/z= 365 (M+H) ⁺ ;

¹ H NMR (DMSO-de): d 10.85 (s, 1 H), 8.31 (s, 1 H), 8.20 (d, J=9.0 Hz, 1 H) ₁ 7.78 (dd, J=9.3 Hz and 1.5 Hz, 1 H), 7.58 (d, J=7.5 Hz, 1 H), 7.24 (d, J=7.5 Hz, 1 H), 7.14-7.04 (m, 2H), 6.21 (s, 1 H), 4.85 (m, 1 H), ) 3.86 (s, 3H), 3.42 (m, 2H); Mp=237-238°C.

2) Second Step: Synthesis of (1 R,3R)-1-Benzo[1 ,2,5]thiadiazol-5-yl-2-(2- chloro-acetyl)-2,3,4,9-tetrahydro-1H-β-carboline-3-carboxyl ic acid methyl ester (Compound 67c)

67c was prepared according to general protocol 2 starting from compound 64c obtained at step 1 above (200 mg, 1 eq.). Compound 67c was obtained as a yellow powder (205 mg, 84%) which was directly engaged in the cyclisation step.

LC: t _R = 6.2 min; MS (ESI+): m/z=439 (M-H)-

3) Third Step: Synthesis of Compound 70c

Compound 70c was prepared according to general protocol 4 starting from compound 67c as obtained above at step 2 (185 mg, 0.42 mmol) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (145 μl, 2 eq.) in MeOH. Compound 70c was obtained after purification by TLC (CH ₂ CI ₂ /MeOH 95:5 v/v) as a yellow powder (120 mg, 44%).

LC: t _R = 5.03 min; MS (ESI+): m/z= 549 (M+H) ⁺ ;

¹ H NMR (DMSO-de): of 11.2 (s, 1 H), ) 8.01 (s, 1 H), 7.97 (d, J=9.30 Hz, 1 H), 7.66 (dd, J=9.0 Hz and 1.5 Hz, 1 H), 7.56 (d, J=7.20 Hz, 1 H),

7.38-7.35 (m, 5H), 7.28 (d, J=7.80 Hz, 1 H), 7.05 (td, J=7.50 Hz and 0.9 Hz,

1 H), 7.02 (td, J=7.80 Hz and 1.2 Hz, 1 H), 6.39 (s, 1 H), 5.07 (m, 1 H), 4.52 (dd,

J=11.4 Hz and 4.2 Hz, 1 H), 4.17 (s, 2H), 3.55 (dd, J=15.9 Hz and 4.5 Hz, 2H),

3.13 (dd, J=15.3 Hz and 11.7 Hz, 2H), 3.12-3.01 (m, 1 H), 2.96-2.80 (m, 1 H), 2.75 (m, 1 H), 2.23 (m, 2H), 2.02-1.90 (m, 1 H).

EXAMPLE 17: Synthesis of (6S,12aR)-6-Benzo[1,2,5]thiadiazol-5-yl-2-((R)- 1-benzyl-pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro-pyrazino[ 1',2':1,6] pyrido[3,4-ib]indole-1,4-dione (Compound 7Ot)

1) First Step: Synthesis of (IS.SRH-Benzoti ^.δlthiadiazol-δ-yl^.SAθ- tetrahydro-1H-β-carboline-3-carboxylic acid methyl ester (Compound 64t)

64t was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (1241 mg) and 2,1 ,3-benzothiadiazole-5-carbaldehyde (800 mg). The reaction mixture was stirred overnight. After filtration and concentration, the cis isomer 64c was separated by precipitation in EtOAc and the residue was purified by column chromatography (CH ₂ CI ₂ /MeOH) to provide the trans compound 64t as a orange solid (160 mg, 10%)

LC: t _R = 4.22 min; MS (ESI+): m/z= 365 (M+H) ⁺ ;

¹ H NMR (DMSO-de): d 10.65 (s, 1 H), 8.03 (d, J=9.3 Hz, 1 H) ₁ 7.80 (s, 1 H), 7.70 (dd, J=9.3 Hz and J=1.8 Hz, 1 H), 7.48 (d, J=7.5 Hz, 1 H), 7.22 (d, J=7.5 Hz, 1 H), 7.06-6.95 (m, 2H), 5.57 (s, 1 H), 3.89 (m, 1 H), 3.62 (s, 3H), 3.07 (m, 2H); Mp=156-160°C.

2) Second Step: Synthesis of (1S,3R)-1-Benzo[1 ,2,5]thiadiazol-5-yl-2-(2- chloro-acetyl)-2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (Compound 67t)

67t was prepared according to general protocol 2 starting from compound 64t as obtained above at step 1 (160 mg, 1 eq.) and obtained as a yellow oil (190 mg, 97%) which was directly engaged in the cyclisation step. LC: t _R = 6.2 min; MS (ESI+): m/z=441 (M+H) ⁺ . 3) Third Step: Synthesis of Compound 7Ot

Compound 7Ot was prepared according to general protocol 4 starting from compound 67t as obtained above at step 2 (190 mg, 0.43 mmol) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (148 μl, 2 eq.) in MeOH. Compound

7Ot was obtained after purification by TLC (CH ₂ CI ₂ /MeOH 95:5 v/v) as a yellow powder (147 g, 60%).

LC: t _R = 5.05 min; MS (ESI+): m/z= 549 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ): δ 11.15 (s, 1 H), 8.10 (d, J=9.6 Hz ₁ 1 H),

7.58 (dd, J=9.3 Hz and 1.5 Hz, 1 H), 7.64 (s, 1 H), 7.56 (d, J=7.5 Hz, 1 H), 7.36 (d, J=8.1 Hz, 1 H), 7.33-7.22 (m, 5H), 7.14 (td, J=7.20 Hz and 1.2 Hz, 1 H) ₁

7.08 (S ₁ 1 H), 7.05 (td, J=7.50 Hz and 0.9 Hz ₁ 1 H) ₁ 5.03-4.95 (m, 1 H), 4.39 (d,

J=17.7 Hz, 1 H), 4.15 (d, J=17.4 Hz, 1 H) ₁ 4.14 (dd, J=12.3 Hz and 4.5 Hz ₁ 1 H) ₁

3.58 (d, J=12.9 Hz, 1 H) ₁ 3.51 (d, J=12.9 Hz ₁ 1 H) ₁ 3.41 (dd, J=15.6 Hz and 4.2

Hz ₁ 1 H) ₁ 3.06 (dd, J=15.0 Hz and 12.3 Hz) 2.86-2.82 (m, 1 H) ₁ 2.72 (dd, J=10.2 Hz and 2.7 Hz ₁ 1 H) ₁ 2.35 (dd, J= 10.2 Hz and 7.8 Hz ₁ 1 H) ₁ 2.20-2.01 (m, 2H),

1.91-1.74 (m, 1 H).

EXAMPLE 18: Synthesis of (6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6-(2,2- difluoro-benzo[1,3]dioxol-5-yl)-2,3,6,7,12,12a-hexahydro-pyr azino- [r,2':1,6]pyrido[3,4-Jb]indole-1,4-dione (Compound 77)

1) First Step: Synthesis of (1 R,3R)-1-(2,2-Difluoro-benzo[1 ,3]dioxol-5-yl)- 2,3,4,9-tetrahydro-1/-/-β-carboline-3-carboxylic acid methyl ester (Compound 73c)

73c was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (1.03 g) and 2,2-difluoro-5-formylbenzodioxole

(930 mg). The reaction mixture was stirred for 17 hours. Compound 73c was obtained after purification by column chromatography, as a white powder

(862 mg, 56%).

LC: t _R =4.6 min; MS (ESI+): m/z=387 (M+H) ⁺ ;

¹ H NMR (DMSO-de), d 10.45 (s, 1H), 7.45 (m, 2H), 7.36 (d, J=1.5 Hz, 1H), 7.25 (Dd, J=1.5 Hz and J=8.3 Hz, 1H), 7.21 (d, J=7.5 Hz, 1H), 7.02 (td, J=I.5 Hz and J=15 Hz, 1H) ₁ 6.95 (td, J= 1.1 Hz and J= 14.5 Hz, 1H),

5.42 (S ₁ 1 H), 4.09 (m, 1 H), 3.75 (s, 3H), 3.10 (m, 1 H), 2.93 (m, 1 H);

Mp = 124-126°C.

2) Second Step: Synthesis of (1 R,3R)-2-(2-Chloro-acetyl)-1-(2,2-difluoro- benzo[1 ,3]dioxol-5-yl)-2,3,4,9-tetrahydro-1 /-/-β-carboline-3-carboxylic acid methyl ester (Compound 75)

75 was prepared according to general protocol 2 starting from compound 73c as obtained above at step 1 (662 mg, 1 eq.). Compound 75 was obtained as an orange powder (783 mg, 100%) which was directly engaged in the cyclisation step.

LC: t _R =7.1 min; MS (ESI+): m/z=383 (M+H) ⁺ . 3) Third Step: Synthesis of Compound 77

Compound 77 was prepared according to general protocol 6 starting from compound 75 as obtained above at step 2 (100 mg, 1 eq.), triethylamine (4 eq.) and 4-amino-N-benzylpiperidine (1.5 eq.) in EtOH.

Compound 77 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (51 mg, 34%).

LC: t _R =5.5 min; MS (ESI+): m/z= 585 (M+H) ⁺ ;

¹ H NMR (DMSO-c/e) ppm: d 11.03 (s, 1 H), 7.54 (d, J= 7.7 Hz,

1 H), 7.33-7.24 (m, 8H), 7.15 (dd, J= 9.9 Hz and 1.5 Hz, 1 H), 7.08-6.97 (m,

2H), 6.19 (s, 1 H), 4.44 (dd, J= 11.7 Hz and 4.3 Hz, 1 H), 4.26 (m, 1 H), 4.03 (d,

J= 17.0 Hz ₁ 1 H), 3.92 (d, J= 16.8 Hz, 1 H), 3.47 (m, 3H) ₁ 3.04 (dd, J= 15.4 Hz and 11.5 Hz, 1 H) ₁ 2.88 (m, 2H), 2.03 (m, 2H), 1.82-1.46 (m, 4H).

EXAMPLE 19: Synthesis of (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6- (6-chloro-pyridin-3-yl)-2,3,6,7,12,12a-hexahydro-pyrazino[1 ^l ,2':1,6] pyrido[3,4-b]indole-1,4-dione (Compound 84c)

11 First Step: Synthesis of (1 R,3R)-1-(6-Chloro-pyridin-3-yl)-2,3A9- tetrahydro-1/-/-β-carboline-3-carboxylic acid methyl ester (Compound 81c)

81c was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (1.02 g) and δ-chloropyridine-S-carboxaldehyde (708 mg), after stirring for 67 hours and purification by column chromatography. Compound 81c was obtained as a white powder (937 mg,

69%).

LC: t _R =3.8 min; MS (ESI+): m/z=342 (M+H) ⁺ ; ¹ H NMR (DMSO-CZ ₆ ): d 10.45 (s, 1 H), 8.42 (d, J=2.1 Hz, 1 H),

7.72 (dd, J=8.3 Hz and J= 2.5 Hz, 1 H), 7.50 (d, J=8.5 Hz ₁ 1 H), 7.44 (d, J=7.0

Hz, 1 H), 7.13 (m, 1 H), 6.98 (m, 2H), 5.3 (d, J=5.5 Hz, 1 H) , 3.89 (m, 1 H), 3.71

(s, 3H), 3.21 (t, J=5.5 Hz, 1 H), 3.05 (dd, J=14.9 Hz and J=3.1 Hz, 1 H), 2.85

(m, 1 H); Mp = 190-192 ⁰ C.

2) Second Step: Synthesis of (1 R,3R)-2-(2-Chloro-acetyl)-1-(6-chloro-pyridin- 3-yl)-2,3,4,9-tetrahydro-1H-β-carboline-3-carboxylic acid methyl ester (Compound 83c)

83c was prepared according to general protocol 2 starting from compound 81c as obtained above at step 1 (681 mg, 1 eq.). Compound 83c was obtained as a white powder (667 mg, 80%) which was directly engaged in the cyclisation step. LC: t _R = 6.0 min; MS (ESI+): m/z=420 (M+H) ⁺ .

3) Third Step: Synthesis of Compound 84c

Compound 84c was prepared according to general protocol 6 starting from compound 83c as obtained above at step 2 (0.44 mmol), triethylamine (123 μl, 2 eq.) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (152 μl, 2 eq.) in MeOH. Compound 84c was obtained after purification by TLC (CH ₂ CI ₂ /MeOH 95:5 v/v) as a white powder (100 mg, 43%). LC: t _R = 4.69 min; MS (ESI+): m/z= 526 (IvHH) ⁺ ;

¹ H NMR (MeOD): of 8.4 (d, J=2.4 Hz ₁ 1 H) ₁ 7.63 (dd, J=2.4 Hz and 8.4 Hz ₁ 1 H), 7.55 (d, J=7.20 Hz, 1 H), 7.37-7.22 (m, 7H), 7.10 (td, J=7.50

Hz and 1.2 Hz, 1 H), 7.04 (td, J=7.20 Hz and 1.2 Hz, 1 H), 6.27 (s, 1 H), 5.12

(m, 1 H), 4.40 (dd, J=11.4 Hz and 4.8 Hz, 1 H), 4.32 (d, J=17.7 Hz, 1 H), 4.12

(dd, J=MA Hz and 1.5 Hz, 1 H), 3.70 (d, J=12.9 Hz, 1 H), 3.67 (dd, J=15.9 Hz and 4.5 Hz, 1 H), 3.54 (d, J=12.6 Hz, 1 H), 3.15 (dd, J=15.9 Hz and 11.4 Hz, 1 H), 2.94 (m, 1 H), 2.75 (dd, J=10.5 Hz and 3.3 Hz, 1 H), 2.48 (dd, J=10.5 Hz and 7.8 Hz, 1 H), 2.34-2.20 (m, 2H), 1.74-1.65 (m, 1 H).

EXAMPLE 20: Synthesis of (6S,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6- (6-chloro-pyridin-3-yl)-2,3,6,7,12,12a-hexahydro-pyrazino[1' ,2':1,6] pyrido[3,4-b]indole-1,4-dione (Compound 84t)

H First Step: Synthesis of (1S,3R)-1-(6-Chloro-pyridin-3-yl)-2,3,4,9- tetrahydro-1 /-/-β-carboline-3-carboxylic acid methyl ester (Compound 81t)

81t was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (1.02 g) and δ-chloropyridine-S-carboxaldehyde (708 mg), after stirring for 67 hours and purification by column chromatography. Compound 811 was obtained as a white powder (262 mg, 19%).

LC: t _R =3.9 min;

MS (ESI+): m/z=342 (M+H) ⁺ ;

¹ H NMR (DMSO-Gf ₆ ), d: 10.50 (s, 1H), 8.35 (d, J=2.3 Hz, 1 H), 7.67 (dd, J=2.4 Hz and J=8.2 Hz, 1 H), 7.44 (m, 2H), 7.22 (d, J=7.9 Hz, 1 H),

7.02 (td, J=1.4 Hz and J=8.2 Hz, 1 H), 6.95 (td, J=1.1 Hz and J=7.5 Hz,1 H),

5.41 (s, 1 H) ₁ 3.82 (m, 1 H), 3.62 (s, 3H), 3.50 (s, 1 H), 3.06 (dd, J=5.5 Hz and

J=15.1 Hz, 1 H), 2.92 (dd, J=5.4 Hz and J=15.1 Hz, 1 H);

Mp = 180-183 ⁰ C.

2) Second Step: Synthesis of (1S,3R)-2-(2-Chloro-acetyl)-1-(6-chloro-pyridin- S-yl^SAθ-tetrahydro-I H-β-carboline-S-carboxylic acid methyl ester (Compound 83t)

83t was prepared according to general protocol 2 starting from compound 81t as obtained above at step 1 (150 mg, 1 eq.). Compound 83t was obtained as a white powder (147 mg, 80%) which was directly engaged in the cyclisation step.

LC: t _R = 5.92 min; MS (ESI+): m/z=418 (M+H) ⁺ .

3) Third step: Synthesis of Compound 84t

Compound 84t was prepared according to general protocol 6 starting from compound 83t as obtained above at step 2 (0.44 mmol), triethylamine (185 μl, 3 eq.) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (227 μl, 3 eq.) in MeOH and obtained after purification by TLC (CH ₂ CI ₂ /MeOH 95:5 v/v) as a white powder (120 mg, 52%). LC: t _R = 4.77 min; MS (ESI+): m/z= 526 (M+H)+;

¹ H NMR (MeOD): d 8.34 (d, J=2.4 Hz ₁ 1 H) 7.65 (dd, J=2.7 Hz and 8.4 Hz, 1 H), 7.52 (d, J=7.8 Hz, 1H), 7.41 (d, J=8.4 Hz, 1 H), 7.36-7.24 (m,

6H), 7.15 (td, J=6.9 Hz and 1.2 Hz, 1 H), 7.07 (td, J=6.9 Hz and 0.9 Hz, 1 H),

7.03 (s, 1 H), 5.12-5.04 (m, 1 H), 4.36 (d, J=17.7 Hz, 1 H), 4.27 (d, J=18.0 Hz,

1 H), 4.23 (dd, J=M J Hz and 4.2 Hz, 1 H), 3.64 (d, J=12.6 Hz, 1 H), 3.58 (d,

J=12.6 Hz, 1 H), 3.42 (dd, J=15.3 Hz and 4.2 Hz, 1 H), 3.04 (dd, J=15.3 Hz and 12.0 Hz, 1 H), 3.02-2.87 (m, 1 H), 2.77 (dd, J=10.5 Hz and 3.3 Hz, 1 H) ₁ 2.50

(dd, J=10.5 Hz and 7.5 Hz, 1 H), 2.33 (m, 1 H), 2.21-2.14 (m, 1 H), 1.98-1.87

(m, 1 H).

EXAMPLE 21 : Synthesis of (6R,12aR)-6-Benzyl-2-(1-benzyl-piperidin-4- yl)-2,3,6,7,12,12a-hexahydro-pyrazino[r,2':1,6]pyrido[3,4-^] indole-1,4- dione (Compound 89c)

1) First Step: Synthesis of (1 R,3R)-1-Benzyl-2,3,4,9-tetrahydro-1 H-β- carboline-3-carboxylic acid methyl ester (Compound 85c)

85c was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (500 mg) and phenylacetaldehyde (274 μl, 1.25 eq.), after stirring for 72 hours and purification by TLC (chloroform/ether). Compound 85c was obtained as a white powder (120 mg, 19%). LC: t _R =3.9 min; MS (ESI+): m/z= 321 (M+H) ⁺ ;

¹ H NMR (CDCI ₃ ) : of 7.50 (d, J=8.0 Hz, 1 H), 7.37 (m, 5H), 7.22-7.08 (m, 4H), 4.55 (t, J=6.8 Hz, 1 H) ₁ 3.83 (s, 3H), 3.79 (dd, J=8.4 Hz and J=4.2 Hz, 1 H), 3.25 (dd, J=13.4 Hz and J=6.3 Hz, 1 H), 3.16 (dd, J=15.0 Hz and J=12.3 Hz, 1 H), 3.06 (dd, J=13.4 Hz and J=7.7 Hz, 1 H), 2.87 (m, 1 H);

¹³ C NMR (CDCI ₃ ), d 173.39, 137.58, 135.68, 134.97, 129.48, 129.09, 127.28, 126.71 , 121.88, 119.60, 118.07, 110.86, 108.38, 56.47, 54.12, 52.31 , 41.67, 25.86.

2) Second Step: Synthesis of (1 R,3R)-1-Benzyl-2-(2-chloro-acetyl)-2,3,4,9- tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (Compound 87c)

87c was prepared according to general protocol 2 starting from compound 85c as obtained above at step 1 (120 nig, 1 eq.). Compound 87c was obtained as a yellow powder (148 mg, 100%) which was directly engaged in the cyclisation step. LC: t _R =6.7 min; MS (ESI+): m/z= 397 (M+H) ⁺ . 3) Thirs Step: Synthesis of Compound 89c

Compound 89c was prepared according to general protocol 6 starting from compound 87c as obtained above at step 2 (148 mg, 1 eq.) and 4-amino-N-benzylpiperfdine (2.5 eq.) in EtOH. Compound 89c was obtained after purification by TLC (chloroform/ether) as a white powder (95 mg, 49%). LC: t _R = 4.9 min;

MS (ESI+): m/z= 519 [IvRH] ⁺ ;

¹ H RMN (CDCI ₃ ), d : 8.06 (s, 1 H), 7.51 (d, J=7.5 Hz, 1 H), 7.33 (m, 6H), 7.23-7.08 (m, 5H), 6.73 (d, J=7.0 Hz, 1 H), 5.61 (t, J=5λ Hz, 1 H), 4.50 (m, 1 H), 4.16 (d, J=16.9 Hz, 1 H), 3.96 (dd, J=11.5 Hz and J=4,9 Hz, 1 H), 3.86 (d, J=16.7 Hz, 1H), 3.63 (s, 2H), 3.29 (dd, J=15.7 Hz and J=5,0 Hz, 1H), 3.18 (d, J=5.13 Hz, 2H), 3.09 (m, 2H), 2.25 (q, J=11 ,8 Hz, 2H), 2.05 (dd, J=15.5 Hz and J=11.4 Hz, 1 H), 1.96-1.70 (m, 4H).

EXAMPLE 22: Synthesis of (6S,12aR)-6-Benzyl-2-(1-benzyl-piperϊdin-4- yl)-2^6JJ2,12a-hexahydro^yrazino[1\2':1,6]pyrido[3Ab]indole- 1,4- dione (Compound 89t)

1) First Step: Synthesis of (1S,3R)-1-Benzyl-2,3,4,9-tetrahydro-1 H-β- carboline-3-carboxylic acid methyl ester (Compound 85t)

85t was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (500 mg) and phenylacetaldehyde (274 μl, 1.25 eq.), after stirring for 72 hours and purification by TLC (chloroform/ether). Compound 85t was obtained as a white powder (70 mg, 11%).

LC: t _R =4.2 min;

MS (ESI+): m/z= 321 (M+H) ⁺ ;

¹ H NMR (CDCI ₃ ): d 7.64-7.30 (m, 6H), 7.15 (m, 4H) ₁ 4.24 (t, J=5,7 Hz, 1 H), 3.83 (m, 1 H), 3.81 (s, 3H), 3,36 (m, 4H);

¹³ C NMR (CDCI ₃ ), d 169.77, 136.05, 135.81 , 129.74, 129.10, 127.67, 125.79, 122.66, 119.86, 118.28, 111.22, 106.04, 53.20, 52.67, 52.51 , 39.68, 22.77.

2) Second Step: Synthesis of (1S,3R)-1-Benzyl-2-(2-chloro-acetyl)-2,3,4,9- tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (Compound 87t)

87t was prepared according to general protocol 2 starting from compound 85t as above obtained at step 1 (70 mg, 1 eq.). Compound 87t was obtained as a yellow powder (85 mg, 99%) which was directly engaged in the cyclisation step.

LC: t _R =6.5 min; MS (ESI+): m/z= 397 (M+H) ⁺ . 3) Third Step: Synthesis of Compound 89t

Compound 89t was prepared according to general protocol 6 starting from compound 87t as above obtained at step 2 (95 mg, 1 eq.) and 4-amino-N-benzylpiperidine (2.5 eq.) in EtOH. and obtained after purification by TLC (chloroform/ether) as a white powder (66 mg, 61 %). LC: t _R = 4.7 min;

MS (ESI+): m/z= 519 [M+H] ⁺ ;

¹ H RMN (CDCI ₃ ): d 7.47 (m, 2H), 7,37-7,07 (m, 12H), 6.04 (dd, J=8.5 Hz and J=5.7 Hz, 1 H), 4.54 (m, 1 H), 4.34 (dd, J=11.7 Hz and J=3.9 Hz, 1 H), 4.03 (d, J=17.6 Hz, 1 H), 3.95 (d, J=MJ Hz, 1 H), 3.59 (s, 2H), 3.46 (dd, J=15.4 Hz and J=4.0 Hz, 1 H), 3.26 (dd, J=13.0 Hz and J=5.6 Hz, 1 H), 3.15-3.03 (m, 3H), 2.84 (dd, J=14.4 Hz and J=3.3 Hz, 1 H), 2.21 (m, 2H), 1.88 (m, 2H), 1.69 (m, 2H).

EXAMPLE 23: Synthesis of (6R,12aR)-6-Benzyl-2-[1-(1-methyl-1H- imidazol-2-ylmethyl)-piperidin-4-yl]-2,3,6,7,12,12a-hexahydr o-pyrazino- [1',2':1,6]pyrido[3,4-Jb]indole-1 ₅ 4-dione (Compound 91c)

1) First Step: Synthesis of a reactive amine: 1-(1-Methyl-1/-/-imidazol-2- ylmethyl)-piperidin-4-yl-ammonium trichloride (compound 59)

Cf

a) Synthesis of [1-(1-Methyl-1H-imidazol-2-ylmethyl)-piperidin- 4-yl]-carbamic acid ferf-butyl ester (Compound 61)

2-(chloromethyl)-1-methyI-1 H-imidazole hydrochloride (800 mg, 1 eq.) and 4-(N-Boc-amino)piperidine (959 mg, 1 eq.) were dissolved in CH ₂ CI ₂ (48 rτiL) and Diisopropyl Ethyl Amine (DIEA) (2.5 mL, 3 eq.) was added. The reaction mixture was stirred at room temperature for 48 h. Solvent was evaporated, the residue dissolved in CH ₂ CI ₂ and washed with saturated NaHCO ₃ aqueous solution and water, dried over MgSO ₄ , and evaporated to dryness to give 1.25 g of a yellow powder (89 %)

LC: t _R =2.6 min;

MS (ESI+): m/z = 295 (M+H) ⁺ ; ¹ H NMR (DMSO-CZ ₆ ) : d 7.05 (s, 1 H), 6.73 (m, 2H), 3.61 (s,

3H), 3.45 (s, 2H), 3.18 (m, 1 H), 2.68 (d, J=11 .8 Hz, 2H), 1.95 (t, J=11 .6 Hz,

2H), 1.64 (d, J=10.7 Hz ₁ 2H) ₁ 1.36 (s, 9H), 1.29 (m, 2H). b) Synthesis of Compound 59

Intermediate 61 as obtained above at step a) (940 mg) was dissolved in a mixture of methanol (16 ml_) and HCI in dioxane 4N (16 mL). The reaction mixture was stirred at 40 ⁰ C for 5h. After evaporation, compound 59 was obtained as a white powder (957 mg, 100%). LC: t _R =0.6 min; MS (ESI+): m/z = 195 (M+H) ⁺ 2) Second Step: Synthesis of Compound 91c Compound 91c was prepared according to general protocol 3 starting from a mixture of compounds 87c and 87t as respectively above obtained at each Step 2 of Examples 21 and 22 (110 mg, 1 eq.) and Compound 59 as above prepared at step 1 ) b) (2.2 eq.) in MeOH. Compound 91c was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (20 mg, 14%).

LC: t _R = 4.4 min; MS (ESI+): m/z= 523 [M+H] ⁺ ;

¹ H RMN (MeOD), d : 7.36 (m, 2H), 7.10 (m, 4H), 7.01 (m, 3H),

6.91 (s, 1 H), 6.47 (d, J=7.7 Hz, 2H), 5.64 (m, 1 H), 4.32 (m, 1 H), 4.07 (m, 3H), 3.80 (s, 3H), 3.66 (s, 2H), 3.50 (dd, J=13.1 Hz and J=5.2 Hz, 1 H), 3.08 (m,

4H), 2.23 (m, 2H), 2.02-1.67 (m, 4H), 1.38 (dd, J=11.45 Hz and J=11.6 Hz,

1 H).

EXAMPLE 24: Synthesis of (6S,12aR)-6-Benzyl-2-[1-(1-methyl-1H- imidazol-2-ylmethyl)-piperidin-4-yl]-2,3 _J 6,7,12,12a-hexahydro-pyrazino- [1 ^> ,2':1,6]pyrido[3,4-ϋ]indole-1,4-dione (Compound 91t)

91t

Compound 911 was prepared according to general protocol 3 starting from a mixture of compounds 87c and 87t as respectively above obtained at each Step 2 of Examples 21 and 22 (110 mg, 1 eq.) and Compound 59 as above obtained at Step 1 ) b) of Example 23 (2.2 eq.) in MeOH. Compound 911 was obtained after purification by TLC (CH ₂ CI ₂ ZMeOH) as a white powder (30 mg, 21 %).

LC: t _R = 4.3 min;

MS (ESI+): m/z= 523 [M+H] ⁺ ;

¹ H RMN (CDCI ₃ ): d 7.63 (dd, J=10.6 Hz and J=1.7 Hz, 2H), 7,39 (m, 2H), 7.23-7.11 (m, 6H), 7.03 (t, J=7.1 Hz, 1 H), 6.01 (t, J=6.8 Hz, 1 H), 4.42 (m, 1 H), 4.30 (s, 2H), 4.13-3.89 (m, 6H) ₁ 3.41-3.19 (m, 3H), 3.59 (s, 2H), 2.87 (m, 3H), 2.11 (m, 2H), 1.84 (m, 2H).

EXAMPLE 25: Synthesis of (6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6- phenyl-2,3,6,7,12,12a-hexahydro-pyrazino[r,2':1,6]pyrido[3,4 -b]indole- 1,4-dione (Compound 90)

1) First Step: Synthesis of (1 R,3R)-1-Phenyl-2,3,4,9-tetrahydro-1 H-β- carboline-3-carboxylic acid methyl ester (Compound 86c)

86c was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (1.02 g) and 4-benzaldehyde (507 μl), after

stirring for 48 hours and purification by column chromatography. Compound 86c was obtained as a white powder (716 mg, 58%).

LC: t _R =3.6 min;

MS (ESI+): m/z=307 (M+H) ⁺ ; ¹ H NMR (DMSO-de), d 10.34 (s, 1 H), 7.42 (d, J=8.0 Hz, 1 H),

7.36 (m, 5H), 7.19 (d, J=7.3 Hz, 1 H), 6.92 (m, 2H), 5.21 (d, J= 5.1 Hz, 1 H), 3.84 (m, 1 H), 3.70 (s, 3H), 3.01 (dd, J= 2.3 Hz and J= 13.3 Hz, 1 H), 2.79 (m, 1 H), 2.69 (t, J= 4.9 Hz, 1 H);

Mp = 223-225°C. 2) Second Step: Synthesis of (1 R,3R)-2-(2-Chloro-acetyl)-1-phenyl-2,3,4,9- tetrahydro-1/-/-β-carboline-3-carboxylic acid methyl ester (Compound 88)

88 was prepared according to general protocol 2 starting from compound 86c as obtained above at step 1 (516 mg, 1 eq.). Compound 88 was obtained as a white powder (552 mg, 92%) which was directly engaged in the cyclisation step.

LC: t _R =6.6 min;

MS (ESI+): m/z=383 (IVHH) ⁺ . 3) Third Step: Synthesis of Compound 90 Compound 90 was prepared according to general protocol 6 starting from compound 88 as obtained above at step 2 (100 mg, 1 eq.) and 4-amino-N-benzylpiperidine (2.2 eq.) in MeOH. Compound 90 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (74 mg, 56%).

LC: t _R = 4.7 min; MS (ESI+): m/z= 505 [M+H] ⁺ ;

¹ H NMR (DMSO-de), d (ppm) : 11.14 (s, 1 H), 7.53 (d, J= 7.5 Hz ; 1 H), 7.22 (m, 10H), 7.15 (m, 1 H), 6.96 (m, 2H), 6.25 (s, 1 H), 4.43 (dd, J ₁ = 4.6 Hz ₁ J ₂ = 11.5 Hz ; 1 H), 4.23 (m, 1 H), 4.04 (d, J=16.6 Hz, 1 H), 3,87 (d,

J=16.7 Hz ₁ 1 H), 3,46 (m, 3H) ₁ 2.86 (m, 3H), 2.02 (m, 2H), 1.65 (m, 2H), 1.56

(m, 1 H), 1.45 (m, 1 H).

EXAMPLE 26: Synthesis of 6-Benzo[1 ,3]dioxol-5-yl-2-(1-benzyl-piperidin-

4-yl)-9-fluoro-2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1, 6]pyrido[3,4-b]- indole-1,4-dione (CIS : (1 R,3R) and (1S,3S), (as a mixture of (6R, 12aR) and (6S, 12aS) stereoisomers) (Compound 95)

1) First Step: Synthesis of 2-(6-Fluoro-1 H-indol-3-yl)-1-methoxycarbonyl- ethyl-ammonium chlohydrate (Compound 92)

Compound 92 was prepared according to general protocol 5 starting from 6-fluoro-DL-tryptophan (4.67 g, 250 mmol) and obtained as a white solid (5.7 g, 100 %).

LC: t _R = 3.2 min; MS (ESI+): m/z= 237 (M+H) ⁺ ;

¹ H NMR (DMSO-c/ ₆ ): of 11 ,24 (s, 1 H), 7,51 (dd, J=8,7 Hz and J=5,4 Hz, 1 H), 7,26 (d, J=2,3 Hz, 1 H), 7,15 (dd, J=10,1 Hz and J=2,3 Hz, 1 H), 6,87 (ddd, J=11 ,0 Hz, J=8,7 Hz and J=2,4 Hz, 1 H), 4,21 (t, J=6,4 Hz, 1 H), 3,64 (s, 3H), 3,30 (m, 2H).

2) Second Step: Synthesis of 1-Benzo[1 ,3]dioxol-5-yl-7-fluoro-2,3,4,9- tetrahydro-1H-β-carboline-3-carboxylic acid methyl ester (CIS : mixture of enantiomers (1 R,3R) and (1 S,3S)) (Coumpound 93)

93 was prepared according to general protocol 1 starting from compound 92 as obtained above at step 1 (1.4 g) and piperonal (940 mg), after stirring for 120 hours and purification by column chromatography. Compound 93 was obtained as a white powder (590 mg, 32%).

LC: t _R = 4.0 min;

MS (ESI+): m/z= 369 (M+H) ⁺ ;

¹ H NMR (DMSO-de): d 10.42 (s, 1 H), 7.42 (dd, J=8.5 Hz and J=5.5 Hz, 1 H), 6.97-6.77 (m, 5H), 6.00 (s, 2H), 5.11 (d, J=5.4 Hz, 1 H), 3.84 (dt, J=11.1 Hz and J=AA Hz, 1 H), 3.70 (s, 3H), 3.00 (dd, J=15.1 Hz and J=2.88 Hz, 1 H), 2.85-2.69 (m, 2H).

3) Third Step: Synthesis of 1-Benzo[1 ,3]dioxol-5-yl-2-(2-chloro-acetyl)-7- fluoro-2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (CIS : (1 R,3R) and (1S,3S), mixture of enantiomers) (Compound 94)

94 was prepared according to general protocol 2 starting from compound 93 as obtained above at step 2 (378 mg) and obtained as a yellow powder (392 mg, 86%) which was directly engaged in the cyclisation step.

LC: t _R = 6.4 min;

MS (ESI+): m/z=445 (M+H) ⁺ . 4) Forth Step: Synthesis of Compound 95

95 was prepared according to general protocol 6 starting from

94 as obtained above at step 3 (53 mg) and 1-Benzyl-pipehdin-4-ylamine (57 μl) in 2 minutes. Compound 95 was obtained as a white powder (27 mg, 43%).

LC: t _R = 4.8 min;

MS (ESI+): m/z = 567 (M+H) ⁺ ;

¹ H NMR (CDCI ₃ ) : d 7,62 (m, 1 H), 7,47 (m, 5H), 6,91 (m, 2H), 6,76 (m, 2H), 6,48 (d, J=8,4 Hz, 1 H), 6,25 (s, 1 H), 5,85 (d, J=1 ,4 Hz, 1 H), 5,84 (d, J=1 ,4 Hz, 1 H), 4,61 (m, 1 H), 4,31 (dd, J=11 ,6 Hz and J=4,8Hz, 1 H), 4,29 (m, 1 H), 4,07 (m, 2H), 3,90 (d, J=16,9 Hz, 1 H), 3,68 (m, 1 H), 3,60 (dd, J=15,7 Hz and J=4,4 Hz ₁ 1 H), 3,43 (m, 1 H), 3,12 (dd, J=15,9 Hz and J=12,1 Hz, 1 H), 2,68 (m, 4H), 1 ,98 (d, J= 11 ,9 Hz, 1 H), 1 ,70 (m, 1 H).

EXAMPLE 27: Synthesis of (6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6-(4- difluoromethoxy^henyI)-2^6J,12,12a-hexahydro-pyrazino[1\2':1 ,6] pyrido[3,4-b]indole-1,4-dione (Compound 96)

1) First Step: Synthesis of (1 R,3R)-1-(4-Difluoromethoxy-phenyl)-2,3,4,9- tetrahydro-1/-/-β-carboline-3-carboxylic acid methyl ester (Compound 74c)

74c was prepared according to general protocol 1 starting from D-tryptophan-OMe.HCI (1.02 g) and 4-(difluoromethoxy)benzaldehyde

(862 mg), after stirring for 72 hours and purification by column chromatography. Compound 74c was obtained as a white powder (845 mg,

57%).

LC: t _R =4.3 min;

MS (ESI+): m/z=373 (M+H)+;

1H NMR (DMSO-c/6), d : 10.36 (s, 1 H), 7.41 (d, J=8.4 Hz, 2H), 7.23 (t, JH-F= 74 Hz, 1 H), 7.21 (m, 1 H), 7.15 (d, J=8.4 Hz, 2H), 6.98 (m, 2H), 5.23 (s, 1 H) ₁ 3.89 (m, 1 H), 3.72 (s, 3H), 3.02 (dd, J=2.6 Hz and J= 13.8 Hz, 1 H), 2.83 (dd, J= 13 Hz and J= 14.6 Hz, 1 H);

Mp = 90-93 ⁰ C.

2) Second Step: Synthesis of (1 R,3R)-2-(2-Ch!oro-acetyl)-1-(4- difluoromethoxy-phenyl)-2,3,4,9-tetrahydro-1 /-/-β-carboline-3-carboxylic acid methyl ester (Compound 76)

76 was prepared according to general protocol 2 starting from compound 74c as obtained above at stepi (625 mg, 1 eq.) and obtained as an orange powder (711 mg, 94%) which was directly engaged in the

cyclisation step.

LC: t _R =6.8 min;

MS (ESI+): m/z=449 (M+H) ⁺ . 3) Third Step: Synthesis of Compound 96 Compound 96 was prepared according to general protocol 6 starting from compound 76 as obtained above at step 2 (50 mg, 1 eq.) and 4-amino-N-benzylpiperidine (2.5 eq.) in EtOH. Compound 96 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (28 mg, 45%)

LC: t _R = 5.09 min; MS (ESI+): m/z= 571 [M+H] ⁺ ;

¹ H RMN (DMSO-de), d: 11.11 (s, 1 H), 7.56 (d, J=7.56 Hz, 2H),

7.37-7.27 (m, 7H), 7.12-6.88 (m, 5H), 6.22 (s, 1 H), 4.45 (dd, J= 11.49 Hz and

J=4λ4 Hz, 1 H), 4.27 (m, 1 H), 4.06 (d, J=16.56 Hz, 1 H), 3.96 (d, J= 16.56 Hz,

1 H), 3.53-3.47 (m, 3H), 3.03 (m, 3H), 1.99 (m, 2H), 1.72 (m, 2H), 1.57 (m, 1 H), 1.48 (m, 1 H).

EXAMPLE 28: Synthesis of (6R,12aR)-2-(1-Benzyl-piperidin-4-yl)-6-(6- chloro-pyridin-3-yl)-2,3,6,7,12,12a-hexahydro-pyrazino[1",2' :1,6]pyrido [3,4-b]indoIe-1,4-dione (Compound 97)

Compound 97 was prepared according to general protocol 6 starting from compound 83c as obtained at step 2 of above Example 19 (50 mg, 1 eq.) and 4-amino-N-benzylpiperidine (2.5 eq.) in ethanol. Compound 97 was obtained after purification by TLC (dichloromethane/methanol) as a white powder (35 mg, 54%) LC: t _R = 4.62 min;

MS (ESI+): m/z= 540 [M+H] ⁺ ;

¹ H RMN (DMSO-de), d: 11.19 (s, 1 H), 8.44 (d, J=2.25 Hz, 1 H),

7.67-7.58 (m, 2H), 7.38-7.24 (m, 7H), 7.09-6.98 (m, 2H), 6.08 (s, 1 H), 4.45

(dd, J= 11.19 Hz and J=4.2, 1 H), 4.26 (m, 1 H), 4.09 (d, J=9.81 Hz ₁ 1 H) ₁ 4.00 (d, J=9.81 Hz, 1 H), 3.53-3.47 (m, 3H), 3.06 (m, 1 H), 2.87 (m, 2H), 2.07 (m,

2H), 1.83-1.44 (m, 4H).

EXAMPLE 29: Synthesis of (6R,12aR)-2-{(R)-1-[4-(2-{2-[2-(2-Amino- ethoxy)-ethoxy]-ethoxy}-ethoxy)-benzyl]-pyrrolidin-3-yl}-6- benzo[1,3]dioxol-5-yl-2,3,6,7,12,12a-hexahydro- pyrazino[1\2':1 ,6]pyrido[3,4-b]indole-1,4-dione (Compound 100)

100

1) First Step: Synthesis of the intermediate (6R,12aR)-6-Benzo[1 ,3]dioxol-5- yl-2-((R)-1 -{4-[2-(2-{2-[2-(1 ,3-dioxo-1 ,3-dihydro-isoindol-2-yl)-ethoxy]-ethoxy}- ethoxy)~ethoxy]-benzyl}-pyrrolidin-3-yl)-2, 3,6,7,12,12a-hexahydro- pyrazino[1',2 ^l :1 ,6]pyrido[3,4-b]indole-1 ,4-dione (148)

148 was prepared according to general protocol 4 starting from intermediate 23 (490 mg) and 4-[2-(2-{2-[2-(1 ,3-Dioxo-1 ,3-dihydro- isoindol-2-yl)-ethoxy]-ethoxy}-ethoxy)-ethoxy]-benzaldehyde (1.1 eq.).

Compound 148 was obtained after purification by TLC (CH ₂ CI ₂ ZMeOH) as a white powder (320mg, 34%).

LC : t _R =5.45min

MS (ESI+): m/z = 856 [M+H] ⁺ . 2) Second Step:

148 (400 mg, 1 eq.) and hydrazine monohydrate (113 μl, 5eq.) were dissolved in ethanol. The reaction mixture was refluxed 3 hours. Ethanol was removed by evaporation and the mixture was diluted with ethyl acetate.

100 was obtained after purification by flash chromatography (CH ₃ CI/MeOH) as a white powder (210 mg, 62%).

LC: t _R =3.76

MS (ESI+): m/z = 726 [M+H] ⁺ .

EXAMPLE 30: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[3- (2-dimethylamino-ethoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6,7,12 ,12a- hexahydrO-pyrazino[1\2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 101)

101 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 3-(2-Dimethylamino-ethoxy)-benzaldehyde (1.5 eq.). Compound 101 was obtained after purification by TLC (CH ₂ CI ₂ /Me0H) as a white powder (38 mg, 27%). LC: t _R =3.89 min; MS (ESI+): m/z = 311 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) : d 11 ,10 (s, 1 H), 7.53 (d, J=7,8 Hz ₁ 1 H), 7.30 (d, J=8.1 Hz, 1 H), 7.20 (m, 1 H), 7.08 (m, 2H), 6.87 (m, 2H), 6.80 (m, 3H),

6.60 (d, J=8,1 Hz, 1 H), 6.18 (s, 1 H), 5.92 (s, 2H) ₁ 5.02 (m, 1 H), 4.40 (m, 1 H), 4,17 (m, 2H), 4.01 (m, 2H), 3.60 (d, J=13.5 Hz, 1 H), 3.50-3.21 (m, 5H), 2,95 (m, 2H), 2,72 (m, 1 H), 2.38 (m, 1 H), 2.28-2,09 (m, 6H), 1.97 (s, 2H), 1.51 (m, 1 H).

EXAMPLE 31 : Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[4- (2-dimethylamino-ethoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6,7, 12,12a- hexahydro-pyrazinoti'^'ri^lpyridotS^-bJindole-i^-dione (Compound 102)

102 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 4-(2-Dimethylamino-ethoxy)-benzaldehyde (1.5 eq.). Compound 102 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (45 mg, 30%).

LC: t _R =3.81 min;

MS (ESI+): m/z = 311 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) : d 11 ,09 (s, 1 H), 7.53 (d, J=7,8 Hz, 1 H), 7.30 (d, J=7,8 Hz, 1 H), 7.20 (d, J=8.7 Hz, 2H), 7.08-6.96 (m, 2H), 6.87 (m, 2H) ₁ 6.80 (m, 2H), 6.70 (dd, J=8.1 Hz and J=1.8 Hz, 1 H), 6.18 (s, 1H), 5.92 (s, 2H) ₁ 5.02 (m, 1 H), 4.40 (m, 1 H), 4,14 (m, 2H), 4.01 (m, 2H), 3.54 (d, J=12.6 Hz, 1 H), 3.50-3.21 (m, 3H), 2,95-2.70 (m, 2H), 2,72 (m, 2H), 2.38 (m, 1 H), 2.28-2,09 (m, 6H), 1.90 (s, 2H), 1.51 (m, 1 H).

EXAMPLE 32: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-((R)-1-{4- [3-(4-methyl-piperazin-1-yl)-propoxy]-benzyl}-pyrrolidin-3-y l)- 2,3,6,7 ₅ 12,12a-hexahydro-pyrazino[1 ^l ,2 ^I :1,6]pyrido[3,4-b]indole-1,4-dione (Compound 104)

104 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 4-[3-(4-Methyl-piperazin-1-yl)-propoxy]- benzaldehyde (1.5 eq.). Compound 104 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (77 mg, 48%). LC: t _R =3.62 min;

MS (ESI+): m/z = 347 (M+H) ⁺ ;

¹ H NMR (DMSO-de) : d 11 ,09 (s, 1 H), 7.53 (d, J=7,5 Hz, 1 H), 7.31 (d, J=7.5 Hz, 1 H), 7.19 (d, J=8.7 Hz ₁ 2H), 7.00 (m, 2H), 6.87-6.76 (m, 5H), 6.17 (s, 1 H), 5.91 (s, 2H) ₁ 5.01 (m, 1 H), 4.40 (dd, J=4.7Hz and 11.3Hz ₁ 1 H), 4.13 (m, 2H), 3.95 (m, 2H), 3.52 (d, J=12.9 Hz, 1 H), 3.45-3.20 (m, 3H) ₁ 3.00-2.80 (m, 4H) ₁ 2,72 (m, 4H), 2.40-2.13 (m, 10H), 1.82 (m, 2H) ₁ 1.51 (m, 1 H).

EXAMPLE 33: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[3- (2-morpholin-4-yl-ethoxy)-benzyl]-pyrroIidin-3-yl}-2,3,6,7, 12,12a- hexahydro-pyrazino[1',2':1 ,6]pyrϊdo[3,4-b]indole-1,4-dione (Compound 105)

105 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 3-(2-Morpholin-4-yl-ethoxy)-benzaldehyde (1.5 eq.). Compound 105 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (62 mg, 42%). LC: t _R =3.81 min;

MS (ESI+): m/z = 333 (M+H) ⁺ ;

¹ H NMR (DMSO-de) : d 11 ,09 (s, 1 H), 7.53 (d, J=I, Q Hz, 1 H),

7.31 (d, J=8.1 Hz, 1 H), 7.21 (m, 1 H), 7.00 (m, 2H), 6.90-6.69 (m, 6H), 6.18 (s,

1 H), 5.93 (s, 2H), 5.01 (m, 1 H), 4.40 (dd, J=4.8Hz and 11.5Hz, 1 H), 4,20 (m, 2H), 4.05 (m, 2H), 3.65-3.30 (m, 8H), 3.00-2.80 (m, 2H), 2,72 (m, 4H), 2.50

(m, 2H), 2.40-2.13 (m, 3H), 1.51 (m, 1 H).

EXAMPLE 34: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[4- (2-morpholin-4-yl-ethoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6,7, 12,12a- hexahydro-pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 106)

106 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 4-(2-Morpholin-4-yl-ethoxy)-benzaldehyde (1.5 eq.). Compound 106 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (50 mg, 34%). LC: t _R =3.81 min;

MS (ESI+): m/z = 333 (M+H) ⁺ ;

¹ H . NMR (DMSO-d ₆ ) : d 11 ,09 (s, 1 H), 7.53 (d, J=7,8 Hz, 1 H),

7.31 (d, J=8.1 Hz, 1H), 7.21 (m, 2H), 7.00 (m, 2H), 6.89-6.68 (m, 5H), 6.18 (s,

1 H), 5.93 (s, 2H), 5.01 (m, 1 H), 4.40 (dd, J=4.8Hz and 11.5Hz, 1 H), 4,13-4.00 (m, 4H), 3.60-3.30 (m, 8H), 3.00-2.80 (m, 2H), 2,72 (m, 4H), 2.50 (m, 2H),

2.40-2.13 (m, 3H), 1.51 (m, 1 H).

EXAMPLE 35: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[4- (3-morpholin-4-yl-propoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6,7, 12,12a- hexahydro-pyrazino[1 ',2':1 ,6]pyrido[3,4-b]indole-1 ,4-dione (Compound 107)

107 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 4-(3-Morpholin-4-yl-propoxy)- benzaldehyde (1.5 eq.). Compound 107 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (76 mg, 46%). LC: t _R =3.78 min;

MS (ESI+): m/z = 340 (M+H) ⁺ ;

¹ H NMR (DMSO-de) : d 11 ,11 (s, 1 H), 7.53 (d, J=7,8 Hz, 1 H) ₁

7.30 (d, J=QA Hz, 1 H) ₁ 7.21 (m, 2H), 7.08-6.96 (m, 2H), 6.88-6.70 (m, 5H) ₁

6.18 (s, 1 H) ₁ 5.92 (s, 2H), 5.01 (m, 1 H), 4.40 (dd, J=4.8Hz and 11.4Hz, 1 H) ₁ 4.13 (S ₁ 2H) ₁ 3.98 (m, 2H) ₁ 3.60-3.30 (m, 8H) ₁ 3.00-2,72 (m, 4H), 2.40-2.13 (m,

3H), 1.95 (m, 6H), 1.51 (m, 1 H).

EXAMPLE 36: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[4- (3-piperidin-1-yl-propoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6,7, 12,12a- hexahydro-pyrazinoπ'^'il^lpyridoβ^-blindole-i ^-dione (Compound 108)

108 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 4-(3-Piperidin-1-yl-propoxy)-benzaldehyde (1.5 eq.). Compound 108 was obtained after purification by TLC (CH ₂ Cl ₂ /MeOH) as a white powder (75 mg, 58%). LC: t _R =3.96 min;

MS (ESI+): m/z = 338 (M+H) ⁺ ;

¹ H NMR (DMSO-de) : d 11 ,11 (s, 1 H), 7.53 (d, J=7,8 Hz, 1 H),

7.31 (d, J=8.1 Hz, 1 H), 7.19 (m, 2H), 7.00 (m, 2H), 6.90-6.68 (m, 5H), 6.18 (s,

1 H), 5.92 (s, 2H), 5.01 (m, 1 H), 4.40 (dd, J=4.8Hz and 11.5Hz, 1 H), 4,13 (m, 2H), 3.95 (m, 2H), 3.59-3.39 (m, 3H), 3.00-2.80 (m, 2H), 2,72 (m, 2H), 2.40-

2.13 (m, 8H), 1.83 (m, 2H), 1.53 (m, 6H), 1.51 (m, 1 H).

EXAMPLE 37: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[4- (3-dimethylamino-propoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6,7,1 2,12a- hexahydro-pyrazinoli'^'il^lpyridoJS^-blindole-ijφdione (Compound 109)

109 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 4-(3-Dimethylamino-propoxy)- benzaldehyde (1.5 eq.). Compound 109 was obtained after purification by TLC (CH ₂ CI ₂ /Me0H) as a white powder (50 mg, 35%). LC: t _R =3.78 min;

MS (ESI+): m/z = 319 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) : d 11 ,09 (s, 1 H), 7.53 (d, J=7,8 Hz, 1 H), 7.30 (d, J=7,8 Hz, 1 H), 7.20 (d, J=8.7 Hz, 2H), 7.08-6.96 (m, 2H), 6.87-6.68 (m, 5H), 6.18 (s, 1 H), 5.92 (s, 2H), 5.01 (m, 1 H), 4.40 (dd, J=4.8Hz and 11.4Hz, 1 H), 4,14 (m, 2H), 3.95 (m, 2H), 3.54 (d, J=12.6 Hz, 1 H), 3.50-3.25 (m, 2H), 2,95-2.70 (m, 2H), 2,67 (m, 1 H), 2.35-2.10 (m, 10H), 1.90 (s, 1 H), 1.84 (m, 2H), 1.51 (m, 1 H).

EXAMPLE 38: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[3- (2-methoxy-ethoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6,7,12,12a-h exahydro- pyrazinoII'^'iljβjpyridoJS^-blindole-i ^-dione (Compound 110)

110 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 3-(2-Methoxy-ethoxy)-benzaldehyde (1.5 eq.). Compound 110 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (84 mg, 62%). LC: t _R =5.01 min; MS (ESI+): m/z = 609 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) : d 11 ,09 (s, 1 H), 7.53 (d, J=7,5 Hz, 1 H), 7.31 (d, J=7.8 Hz, 1 H), 7.24 (m, 1 H), 7.08-6.96 (m, 2H), 6.87 (m, 2H), 6.80 (m, 3H), 6.70 (dd, J=1.8 Hz and 8.1 Hz, 1H), 6.18 (s, 1 H), 5.92 (s, 2H), 5.03 (m, 1 H), 4.40 (dd, J=6.3 Hz and 11.4Hz, 1 H) ₁ 4.17 (m, 2H), 4.05 (m, 2H), 3.63 (m, 2H), 3.47-3.30 (m, 6H), 3.00-2.80 (m, 2H), 2,72 (m, 1 H), 2.37 (m, 1 H), 2.20 (m, 2H), 1.51 (m, 1 H).

EXAMPLE 39: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1-[4- (2-methoxy-ethoxy)-benzyI]-pyrrolidin-3-yl}-2,3,6,7,12,12a-h exahydro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 111)

111 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 4-(2-Methoxy-ethoxy)-benzaldehyde (1.5 eq.). Compound 111 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (75 mg, 55%). LC: t _R =5.01 min; MS (ESI+): m/z = 609 (M+H) ⁺ ;

¹ H NMR (DMSO-de) : d 11 ,10 (s, 1 H), 7.53 (d, J=7,4 Hz ₁ 1 H) ₁ 7.31 (d, J=7,8 Hz ₁ 1 H) ₁ 7.20 (m, 2H) ₁ 7.10-6.96 (m, 2H) ₁ 6.89-6.72 (m, 5H), 6.18 (s, 1 H) ₁ 5.93 (s, 2H) ₁ 5.01 (m, 1 H), 4.40 (dd, J=4.8Hz and 11.7Hz ₁ 1H) ₁ 4,20-4.00 (m, 4H) ₁ 3.64-3.30 (m, 6H) ₁ 3.00-2.80 (m, 2H) ₁ 2,68 (m, 1 H) ₁ 2.35- 2.10 (m, 3H) ₁ 1.90 (s, 2H) ₁ 1.51 (m, 1 H).

EXAMPLE 40: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-{(R)-1- [3,4-bis-(2-methoxy-ethoxy)-benzyI]-pyrroHdin-3-yl}-2 _s 3,6,7,12,12a- hexahydro-pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 112)

112 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 3,4-Bis-(2-methoxy-ethoxy)-benzaldehyde (1.5 eq.). Compound 112 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (90 mg, 59%). LC: t _R =4.82 min;

MS (ESI+): m/z = 683 (M+H) ⁺ ;

¹ H NMR (DMSO-de) : d 11 ,10 (s, 1 H), 7.53 (d, J=7,5 Hz ₁ 1 H),

7.29 (d, J=7,8 Hz, 1 H), 7.10-6.87 (m, 4H), 6.85-6.68 (m, 4H), 6.18 (s, 1 H),

5.93 (s, 2H), 5.01 (m, 1 H), 4.40 (dd, J=4.8Hz and 11.5Hz, 1 H), 4,20-4.00 (m, 6H), 3.61 (m, 5H), 3.50-3.25 (m, 6H), 3.00-2.80 (m, 2H), 2,68 (m, 1 H), 2.35-

2.10 (m, 3H), 1.90 (s, 2H) ₁ 1.51 (m, 1 H).

EXAMPLE 41 : Synthesis of (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6- (2,2-difluoro-benzo[1,3]dioxol-5-yl)-2,3,6,7,12,12a-hexahydr o- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 113)

Compound 113 was prepared according to general protocol 6 starting from compound 75 as obtained above (101 mg, 1 eq.), and 4-amino- N-benzylpiperidine (2.2 eq.) in EtOH. Compound 113 was obtained after purification by Flash chromatography (CHaC^/MeOH) as a white powder (60 mg, 48%). LC : t _R =5.4 min;

MS (ESI+): m/z= 571 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) ppm: d 11.06 (s, 1 H) ₁ 7.55 (d, J=7.95 Hz, 1 H) ₁ 7.31 (m, 8H), 7.13 (dd, J=8.46 Hz and J=1.68 Hz, 1 H), 7.07 (t, J=7λ7 HTL, 1 H), 7.00 (t, J=7.68 Hz, 1 H), 6.21 (s, 1 H), 5.06 (m, 1 H), 4.42 (dd, J=11.4 Hz and J=4.77 Hz ₁ 1 H), 4.15 (s, 2H), 3.45-3.64 (m, 3H), 3.05 (dd, J=14.9 Hz and J=11.0 Hz, 1 H), 2.88 (m, 1 H), 2.70 (m, 1 H), 2.38 (m, 1H), 2.19 (m, 2H), 1.59 (m, 1 H).

EXAMPLE 42: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[1-(1- methyl-1H-imidazoI-2-ylmethyl)-piperidin-4-yl]-2 _J 3,6,7,12,12a-hexahydro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 114)

Compound 114 was prepared according to general protocol 4 starting from compound 5 as obtained above (100 mg, 1 eq.), and compound 59 (1.5 eq.) and triethylamine (4 eq.) in MeOH. Compound 114 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (32 mg, 25%). LC: t _R =4.25 min; MS (ESI+): m/z= 553 (IvHH) ⁺ ;

¹ H NMR (DMSO-dβ) ppm: d 11.10 (s, 1 H), 7.54 (d, J=7.7 Hz, 1 H), 7.30 (d, J=7.9 Hz, 1 H), 6.96-7.08 (m, 3H) ₁ 6.67-6.82 (m, 4H), 6.16 (s, 1 H), 5.92 (s, 2H), 4.42 (dd, J=12.3 Hz and J=4.2 Hz, 1 H), 4.25 (m, 1 H), 4.05 (d, J=16.4 Hz, 1 H), 3.89 (d, J=16.9 Hz, 1 H), 3.65 (s, 3H), 3.53 (s, 2H), 3.47 (dd, J=16.0 Hz and J=4.7 Hz, 1 H), 2.82-3.10 (m, 5H), 2.09 (m, 1 H), 1.69 (m, 1 H), 1.59 (m, 1 H), 1.45 (m, 1 H).

EXAMPLE 43: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-((S)-1- benzyl-pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1 ₅ 4-dione (Compound 115)

Compound 115 was prepared according to general protocol 4 starting from compound 5 as obtained above (3.2g), and (s)-(+)-1-Benzyl-3- aminopyrrolidine (2 eq.) in MeOH and obtained after purification by flash chromatography (CH ₂ CI ₂ /MeOH) as a white powder (2g, 50%). LC: t _R = 4.9 min; MS (ESI+): m/z= 535 (M+H)+;

¹ H NMR (CDCI ₃ ) : d 7.91 (s, 1H) ₁ 7.54 (d, 1H, J=6.9 Hz, 1H) ₁ 7.14-7.37 (m, 7H), 6.77 (dd, J=7.8 Hz and J=1.8 Hz, 1 H), 6.65 (d, J=1.8 Hz, 1 H), 6.61 (d, J=7.8 Hz, 1 H), 6.16 (s, 1 H) ₁ 5.81 (d, J=1.2 Hz, 1 H), 5.79 (d, J=I .2 Hz, 1 H), 5.17 (brs, 1 H), 4.40 (d, J=17.4 Hz, 1 H), 4.18 (dd, J=11.7 Hz and J=4.2 Hz, 1 H), 3.93 (d, J=XlH Hz, 1 H), 3.64 (dd, J=16.2 Hz and J=5.1 Hz, 1 H), 3.58 (d, J=12.9 Hz, 1 H), 3.42 (d, J=12.9 Hz, 1 H), 3.15 (dd, J=15.0 Hz and J= 11.4 Hz, 1 H), 2.88 (m, 1 H), 2.67 (dd, J= 10.8 Hz and J=2.4 Hz, 1 H), 2.45 (dd, J=10.8 Hz and J=8.1 Hz, 1 H), 2.17 (m, 2H), 1.66 (m, 2H).

EXAMPLE 44: Synthesis of (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6- (3,4-dimethoxy-phenyl)-2,3,6,7,12,12a-hexahydro- pyrazino[1\2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 116)

1) First step: Synthesis of the intermediates (1 R.3R) and (1 S, 3R)-1-(3,4- Dimethoxy-phenyl)-2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (Compound 137)

137 was prepared according to general protocol 1 starting from D-tryptophan OMe. HCI (637mg) and obtained as 802 mg of a white powder (87%).

LC: t _R =3.98 min and 4.02 min; MS (ESI+): m/z=367 (M+H) ⁺ ;

2) Second step: Synthesis of the intermediate (1 R.3R) and (1S,3R)-2-(2- Chloro-acetyl)-1 -(3,4-dimethoxy-phenyl)-2,3,4,9-tetrahydro-1 H-β-carboline-3- carboxylic acid methyl ester (Compound 138)

138 was prepared according to general protocol 2 starting from intermediate 137 of step 1 (567 mg) and obtained after precipitation in diethyl ether as a white powder (729 mg, 100%). LC: t _R =5.80 min and 5.88 min;

MS (ESI+): m/z = 443 (M+H) ⁺ ; 3) Third Step: Synthesis of Compound 116

Compound 116 was prepared according to general protocol 3 starting from intermediate 138 as obtained here-above in step 2 (100 mg) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (2.2 eq.). Compound 116 was obtained after purification by HPLC as a white powder.

EXAMPLE 45: Synthesis of (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6- phenyl^.S^Jj^^aa-hexahydro-pyrazinoϊi'^^i.δlpyridotS^-blin dole- 1,4-dione (Compound 117)

Compound 117 was prepared according to general protocol 6 starting from compound 88 (100 mg, 1 eq.) and (R)-(-)-1-Benzyl-3- aminopyrrolidine (2.5 eq.) in EtOH. Compound 21 was obtained after purification by TLC (chloroform/ether) as a white powder. LC: t _R =4.74 min;

MS (ESI+): m/z= 491 (M+H) ⁺ ;

EXAMPLE 46: Synthesis of 6-Benzo[1,3]dioxol-5-yl-2-((R)-1-benzyl- pyrrolidin-3-yl)-9-fluoro-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (as a mixture of (6R, 12aR) and (6S, 12aS) stereoisomers) (Compound 118)

Compound 118 was prepared according to general protocol 6 starting from compound 94 (120 mg, 1 eq.) and (R)-(-)-1-Benzyl-3- aminopyrrolidine (2.5 eq.) in EtOH. Compound 118 was obtained after purification by TLC (chloroform/ether) as a white powder.

LC: t _R =5.1 min; MS (ESI+): m/z= 553 (M+H) ⁺ ;

EXAMPLE 47: Synthesis of (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6- (3,4-dihydroxy-phenyl)-2,3,6,7,12,12a-hexahydro- pyrazino[1 ^f ,2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 119)

1) First step: Synthesis of the intermediates (1 R.3R) and (1S, 3R)-1-(3,4- Dihydroxy-phenyl)-2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl

ester (Compound 139)

139 was prepared according to general protocol 1 starting from D-tryptophan OMe. HCI (637 mg) and obtained as 415 mg of a white powder (49%).

LC: t _R =3.63 min; MS (ESI+): m/z=339 (M+H) ⁺ ;

2) Second step: Synthesis of the intermediate (1 R.3R) and (1S,3R)-2-(2- Chloro-acetyl)-1-(3,4-dihydroxy-phenyl)-2,3,4,9-tetrahydro-1 H-β-carboline-3- carboxylic acid methyl ester (Compound 140)

140 was prepared according to general protocol 2 starting from intermediate 139 of step 1 (370 mg). After evaporation, the reaction mixture was dissolved in a mixture Methanol/aqueous NaHCO3 and stirred for 2h00. Compound 140 was extracted to obtain 367 of a red powder (81 %).

LC: t _R =4.89 min and 5.16 min;

MS (ESI+): m/z = 413 (M+H) ⁺ ; 3) Third Step: Synthesis of Compound 119

Compound 119 was prepared according to general protocol 3 starting from intermediate 140 as obtained here-above in step 2 (100 mg) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (2.2 eq.). Compound 119 was obtained after purification by HPLC as a white powder.

EXAMPLE 48: Synthesis of (6R,12aR)-6-Benzo[1 ,3]dioxol-5-yl-2-[(R)-1-(2- methyI-benzyl)-pyrrolidin-3-yl]-2,3, 6,7,12,12a-hexahydro- pyrazinoti'^'il^lpyridotS^-blindole-i^-dione (Compound 120)

120 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and o-Tolualdehyde (1.5 eq.). Compound 120 was obtained after purification by TLC (CH ₂ CI ₂ ZMeOH) as a white powder (80mg, 65%).

LC: t _R =5.39 min; MS (ESI+): m/z = 549 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) : d 11 ,09 (s, 1 H), 7.50 (d, J= 10.5 Hz, 1 H), 7.44 (m, 1 H), 7.30 (d, J= 6.9 Hz, 1 H), 7.19 (m, 1 H), 7.12-6.93 (m, 4H), 6.83- 6.76 (m, 2H), 6.70 (dd, J=2.4Hz and 11.2Hz, 1 H ₁ ), 6.17 (s, 1 H), 5.91 (s, 2H), 5.03 (m, 1 H), 4.40 (dd, J=5.1 Hz and 11.6Hz, 1 H), 4,11 (m, 2H), 3.61 (m, 2H), 3.47-3.28 (m, 3H), 2,99-2.83 (m, 2H), 2.70 (m, 1 H), 2.43 (m, 1 H), 2.21 (m, 2H), 1.51 (m, 1 H).

EXAMPLE 49: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1-(2- chloro-benzyI)-pyrrolidin-3-yl]-2,3,6,7,12,12a-hexahydro- pyrazino[1',2":1,6]pyrido[3,4-b]indole-1,4-dione (Compound 121)

121 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 2-Chlorobenzaldehyde (1.5 eq.). Compound 121 was obtained after purification by TLC (CH ₂ CI ₂ /MeOH) as a white powder (42 mg, 33%).

LC: t _R =5.24 min; MS (ESI+): m/z = 569 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) : d 11 ,09 (s, 1 H), 7.50 (d, J=10.5 Hz, 1 H), 7.44 (m, 1 H), 7.30 (d, J= 6.9 Hz, 1 H), 7.19 (m, 1 H), 7.12-6.76 (m, 6H), 6.70 (dd, J=2.4Hz and 11.2Hz, 1 H), 6.17 (s, 1 H), 5.91 (s, 2H), 5.03 (m, 1 H), 4.40 (dd, J=5.1 Hz and 11.6Hz, 1 H), 4,11 (m, 2H), 3.62 (m, 2H), 3.47-3.28 (m, 3H), 2,99-2.83 (m, 2H), 2.70 (m, 1 H), 2.43 (m, 1 H), 2.21 (m, 2H), 1.54 (m, 1 H).

EXAMPLE 50: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1- (2,6-dichloro-benzyl)-pyrrolidin-3-yI]-2,3,6,7,12,12a-hexahy dro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 122)

122 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 2,6-Dichlorobenzaldehyde (1.5 eq.). Compound 122 was obtained after purification by TLC (CH ₂ CI ₂ /Me0H) as a white powder (37 mg, 28%).

LC: t _R =5.22 min; MS (ESI+): m/z = 603 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) : d 11 ,09 (s, 1 H) ₁ 7.50 (d, J=10.5 Hz ₁ 1H), 7.44 (m, 1 H) ₁ 7.30 (d, J= 6.9 Hz, 1 H), 7.19 (m, 1 H), 7.10-6.76 (m, 5H), 6.70 (dd, J=IAHT. and 11.2Hz, 1 H) ₁ 6.17 (s, 1 H), 5.91 (s, 2H), 5.02 (m, 1 H), 4.38 (dd, J=5.1 Hz and 11.6Hz, 1 H), 4,11 (m, 2H), 3.60 (m, 2H), 3.47-3.28 (m, 3H), 2,99-2.83 (m, 2H), 2.70 (m, 1 H) ₁ 2.43 (m, 1 H), 2.20 (m, 2H) ₁ 1.54 (m, 1 H).

EXAMPLE 51 : Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1- (2,4-difluoro-benzyl)-pyrrol»din-3-yl]-2,3,6,7,12,12a-hexah ydro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 123)

123 was prepared according to general protocol 4 starting from intermediate 23 (100 mg) and 2,4-Difluorobenzaldehyde (1.5 eq.). Compound 123 was obtained after purification by TLC (CHaC^/MeOH) as a white powder (84 mg, 66%).

LC: t _R =5.09 min; MS (ESI+): m/z = 571 (M+H) ⁺ ;

¹ H NMR (DMSO-d ₆ ) : d 11 ,09 (s, 1 H) ₁ 7.50 (d, J=10.5 Hz, 1 H), 7.44 (m, 1 H), 7.30 (d, J= 6.9 Hz, 1 H) ₁ 7.19 (m, 1 H) ₁ 7.10-6.96 (m, 3H) ₁ 6.81- 6.76 (m, 2H), 6.70 (dd, 1 H, J=2.4Hz and 11.2Hz), 6.17 (s, 1 H), 5.91 (s, 2H) ₁ 5.02 (m, 1 H), 4.38 (dd, J=5.1 Hz and 11.6Hz, 1 H), 4,11 (m, 2H), 3.60 (m, 2H), 3.47-3.28 (m, 3H), 2,99-2.83 (m, 2H), 2.70 (m, 1 H), 2.43 (m, 1 H), 2.20 (m, 2H) ₁ 1.54 (m, 1 H).

EXAMPLE 52: Synthesis of 6-Benzo[1,3]dioxol-5-yl-2-((R)-1-benzyl- pyrrolidin-3-yl)-10-fluoro-2,3,6,7,12,12a-hexahydro- pyrazinoII'^M.θlpyridoIS^-blindole-i^-dione (mixture of 6R, 12aR and 6S, 12aS stereoisomers) (Compound 124)

1) First Step: Synthesis of 2-(5-Fluoro-1 H-indol-3-yl)-1-methoxycarbonyl- ethyl-ammonium; chloride (Compound 141 )

Compound 141 was prepared according to general protocol 5 starting from 5-fIuoro-DL-tryptophan (4.67 g, 250 mmol) and obtained as a white solid (5.85 g, 100 %).

LC : t _R = 3.06 min; MS (ESI+): m/z= 237 (M+H) ⁺

2) Second Step: Synthesis of 1-Benzo[1 ,3]dioxol-5-yl-6-fluoro-2,3,4,9- tetrahydro-1H-β-carboline-3-carboxylic acid methyl ester (mixture of 1 R,3R and 1S.3S stereoisomers) (Coumpound 142)

142

142 was prepared according to general protocol 1 starting from compound 141 as obtained above at step 1 (0.5 g) and piperonal (276 mg), after stirring for 120 hours and purification by HPLC. Compound 142 was obtained as a white powder (145 mg, 21 %). LC : t _R = 4.0 min;

MS (ESI+): m/z= 369 (M+H) ⁺ ;

¹ H NMR (DMSO-dβ): d 10.40 (s, 1 H), 7.16-7.21 (m, 2H), 6.79- 6.92 (m, 4H) ₁ 6.00 (s, 2H), 5.13 (brs, 1 H), 3.84 (dd, J=11.0 Hz and J=4.1 Hz, 1 H), 3.70 (s, 3H), 3.00 (ddd, J=14.8 Hz, J=4.1 Hz and J=1.5 Hz, 1 H), 2.78 (ddd, J=16.9 Hz, J=11.1 Hz and J=2.16 Hz, 1H)

3) Third Step: Synthesis of 1-Benzo[1 ,3]dioxol-5-yl-2-(2-chloro-acetyl)-6- fluoro-2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (mixture of 1 R,3R and 1S.3S stereoisomers) (Compound 143)

143 was prepared according to general protocol 2 starting from compound 142 as obtained above at step 2 (100 mg) and obtained as a yellow oil (103 mg, 86%) which was directly engaged in the cyclisation step. LC: t _R = 6.3 min; MS (ESI+): m/z=445 (IVHH) ⁺ . 4) Forth Step: Synthesis of Compound 124

124 was prepared according to general protocol 6 starting from 143 as obtained above at step 3 (103 mg) and (R)-(-)-1-Benzyl-3- aminopyrrolidine (87 μl) in 5 minutes. Compound 124 was obtained as a white powder (50 mg, 46%). LC: t _R = 5.0 min;

MS (ESI+): m/z = 553 (M+H) ⁺ ;

¹ H NMR (DMSO) : d 11.20 (s, 1 H), 7,17-7.36 (m, 7H), 6,88

(td, J=9.33 Hz and J=2.5 Hz, 1 H) ₁ 6,80 (m, 1 H), 6,76 (d, J=7.6 Hz, 1 H), 6,69 (dd, J=7.9 Hz and J=1.3 Hz, 1 H), 6.18 (s, 1 H), 5,92 (m, 2H), 5,03 (m, 1 H), 4,38 (dd, J=11 ,5 Hz and J=4.9 Hz, 1 H), 4,17 (d, J=11 ,1 Hz, 2H), 3.39-3.64 (m, 3H), 2.90 (m, 2H), 2.35-2.72 (m, 2H), 2.08-2.27 (m, 2H), 1.49-1.78 (m, 1 H). EXAMPLE 53: Synthesis of (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6- (3-hydroxy-4-methoxy-phenyl)-2,3,6,7,12,12a-hexahydro- pyrazinoti'^'ii^pyridotS^-blindole-i^-dione (Compound 125)

1) First step: Synthesis of the intermediates (1 R.3R) and (1S, 3R)-1-(3- Hydroxy-4-methoxy-phenyl)-2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (Compound 144)

144 was prepared according to general protocol 1 starting from D-tryptophan OMe.HCI (500 mg) and obtained as 693 mg of a yellow powder (100%).

LC: t _R =3.61 min and 3.71 min; MS (ESI+): m/z=353 (M+H) ⁺ ;

2) Second step: Synthesis of the intermediate (1 R,3R) and (1S,3R)-2-(2- Chloro-acetyl)-1-(3-hydroxy-4-methoxy-phenyl)-2,3,4,9-tetrah ydro-1 H-β- carboline-3-carboxylic acid methyl ester (Compound 145)

145 was prepared according to general protocol 2 starting from intermediate 144 of step 1 (662 mg). After evaporation, the reaction mixture was dissolved in a mixture Methanol/aqueous NaHCO3 and stirred for 2h00. Compound 145 was extracted to obtain 737mg of an orange powder

(91%).

LC: t _R =5.34 min and 5.54 min;

MS (ESI+): m/z = 429 (M+H) ⁺ ; 3) Third Step: Synthesis of Compound 125 Compound 125 was prepared according to general protocol 3 starting from intermediate 145 as obtained here-above in step 2 (100 mg) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (2.2 eq.). Compound 125 was obtained after purification by HPLC as a white powder.

EXAMPLE 54: Synthesis of (6R,12aR)-2-((R)-1-Benzyl-pyrrolidin-3-yl)-6- (4-hydroxy-3-methoxy-phenyI)-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 126)

1) First step: Synthesis of the intermediates (1 R,3R) and (1S ₁ 3R)-1-(4- Hydroxy-3-methoxy-phenyl)-2,3,4,9-tetrahydro-1 H-β-carboline-3-carboxylic acid methyl ester (Compound 146)

146 was prepared according to general protocol 1 starting from D-tryptophan OMe. HCI (637 mg) and obtained as 914 mg of a yellow powder (100%).

LC: t _R =3.67 min;

MS (ESI+): m/z=353 (M+H) ⁺ ;

2) Second step: Synthesis of the intermediate (1 R.3R) and (1S,3R)-2-(2- Chloro-acetyl)-1-(4-hydroxy-3-methoxy-phenyl)-2,3,4,9-tetrah ydro-1 H-β- carboline-3-carboxylic acid methyl ester (Compound 147)

147 was prepared according to general protocol 2 starting from intermediate 146 of step 1 (50 mg). After evaporation, the reaction mixture was dissolved in a mixture Methanol/aqueous NaHCO3 and stirred for

2h00. Compound 147 was extracted to obtain 58mg of an orange powder (100%).

LC: t _R =5.37 min and 5.57 min;

MS (ESI+): m/z = 429 (M+H) ⁺ ; 3) Third Step: Synthesis of Compound 126

Compound 126 was prepared according to general protocol 3 starting from intermediate 147 as obtained here-above in step 2 (58 mg) and (R)-(-)-1-Benzyl-3-aminopyrrolidine (2.2 eq.). Compound 126 was obtained after purification by HPLC as a white powder.

EXAMPLE 55: Synthesis of (6R,12aR)-6-Benzo[1,2,5]thiadiazol-5-yl-2- {(R)-1-[3-(2-methoxy-ethoxy)-benzyl]-pyrroIidin-3-yl}-2,3,6, 7,12,12a- hexahydro-pyrazino[1',2':1,6]pyrϊdo[3,4-b]indole-1,4-dione (Compound 127)

Compound 127 was prepared according to general protocol 6 starting from compound 67c and (R)-1-[3-(2-Methoxy-ethoxy)-benzyl]- pyrrolidin-3-ylamine (2.5 eq.) in EtOH.

EXAMPLE 56: Synthesis of (6R,12aR)-6-Benzo[1,2,5]thiadiazol-5-yl-2- {(R)-1-[3,4-bis-(2-methoxy-ethoxy)-benzyl]-pyrrolidin-3-yl}- 2, 3,6,7, 12,12a- hexahydro-pyrazino[1",2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 128)

Compound 128 was prepared according to general protocol 6 starting from compound 67c and (R)-1-[3,4-Bis-(2-methoxy-ethoxy)-benzyl]- pyrrolidin-3-ylamine (2.5 eq.) in EtOH.

EXAMPLE 57: Synthesis of (6R,12aR)-6-Benzo[1,2,5]thiadiazol-5-yl-2- {(R)-1-[4-(2-methoxy-ethoxy)-benzyl]-pyrrolidin-3-yl}-2,3,6, 7,12,12a- hexahydro-pyrazinoII'^M^lpyridofS^-blindole-i^-dione (Compound 129)

Compound 129 was prepared according to general protocol 6 starting from compound 67c and (R)-1-[4-(2-Methoxy-ethoxy)-benzyl]- pyrrolidin-3-ylamine (2.5 eq.) in EtOH.

EXAMPLE 58: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1-(1- methyl-1-phenyl-ethyl)-pyrrolidin-3-yl]-2,3,6,7,12,12a-hexah ydro- pyrazinoII'^ijθlpyridotS^-blindole-i^-dione (Compound 130)

130 was prepared according from intermediate 23 and (1- Chloro-1-methyl-ethyl)-benzene (1.5 eq.) in acetone in presence of K ₂ CO ₃ .

EXAMPLE 59: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-((R)-1- phenethyl-pyrrolidin-3-yl)-2,3,6,7,12,12a-hexahydro- pyrazino[1',2':1,6]pyrido[3,4-b]indole-1 ,4-dione (Compound 132)

Compound 132 was prepared according to general protocol 6 starting from compound 5 and (R)-1-Phenethyl-pyrrolidin-3-ylamine (2.5 eq.) in EtOH. Compound 132 was obtained after purification by HPLC as a white powder.

LC: t _R =5.23 min;

MS (ESI+): m/z= 549 (M+H) ⁺ ;

EXAMPLE 60: Synthesis of (6R,12aR)-6-Benzo[1,3]dioxol-5-yl-2-[(R)-1-(2- phenoxy-ethyl)-pyrrolidin-3-yl]-2,3,6,7,12,12a-hexahydro- pyrazino[1\2":1,6]pyrido[3,4-b]indole-1,4-dione (Compound 133)

Compound 133 was prepared according to general protocol 6 starting from compound 5 as obtained above at step 2 (100 mg, 1 eq.) and (R)-1-(2-Phenoxy-ethyl)-pyrrolidin-3-ylamine (2.5 eq.) in EtOH.

EXAMPLE 61 : Synthesis of (θR^aaRJ-e-tSaJa-Dihydro-benzoti.Sldioxol- 5-yl)-2-[(R)-1-(1,1-dimethyl-2-phenyl-ethyl)-pyrrolidin-3-yl ]-2,3,6,7,12,12a- hexahydro-pyrazinoti'^'rijGlpyndotS^-blindole-i^-dione (Compound 134)

134 was prepared according to general protocol 4 starting from intermediate 23 and (2-Chloro-2-methyl-propyl)-benzene (1.5 eq.) in acetone in presence of K ₂ CO _3.

EXAMPLE 62: Synthesis of (6R,12aR)-6-(3a,7a-Dihydro-benzo[1,3]dioxol- 5-yl)-2-{1-[3-(2-methoxy-ethoxy)-benzyl]-azetidin-3-yl}-2,3, 6,7,12,12a- hexahydro-pyrazino[1\2':1,6]pyrido[3,4-b]indole-1,4-dione (Compound 135)

Compound 135 was prepared according to general protocol 6 starting from compound 5, and 1-[3-(2-Methoxy-ethoxy)-benzyl]-azetidin-3-yI- ammonium; chloride (2 eq.) in MeOH and obtained after purification by HPLC.

EXAMPLE 63: Synthesis of (6R,12aR)-2-{1-[3,4-Bis-(2-methoxy-ethoxy)- benzyl]-azetidin-3-yl}-6-(3a,7a-dihydro-benzo[1,3]dioxol-5-y l)- 2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1,6]pyrido[3,4-b]ind ole-1,4-dione (Compound 136)

Compound 136 was prepared according to general protocol 6 starting from compound 5, and 1-[3,4-Bis-(2-methoxy-ethoxy)-benzyl]-azetidin- 3-yl-ammonium; chloride (2 eq.) in MeOH and obtained after purification by HPLC. EXAMPLE 64: In vitro demonstration of the antiparasitic activity of compounds of formula (I) 1 ) Material and methods:

Plasmodium cultures

Two clones of P. falciparum are used: (a) the 3D7 clone of NF54 which is known to be sensitive to all anti-malarials, (b) the K1 strain originating from Thailand that is resistant to chloroquine and pyrimethamine, but sensitive to mefloquine. The cultures are naturally asynchronous (65-75% ring stage) and are maintained in continuous log phase growth in RPMI-1640 medium (as developed by Moore et. al. at Roswell Park Memorial Institute, sold by Sigma Aldrich) supplemented with 5% washed human A+ erythrocytes, 25 mM Hepes, 32 nM NaHCO ₃ , and AlbuMAXII (lipid-rich bovine serum albumin) sold GIBCO, Grand Island, NY (CM). All cultures and assays were conducted at 37°C under an atmosphere of 5% CO ₂ and 5% O ₂ , with a balance of N ₂ .

Drug sensitivity assays

Stock drug solutions were prepared using compounds of formula (I) in 100% DMSO (dimethylsulfoxide) at 20 mg/mL unless otherwise suggested by the supplier. The compounds were further diluted to the appropriate concentration using complete medium RPMI-1640 supplemented with 15nM cold hypoxanthine and AlbuMAXII.

Assays were performed in sterile 96-weII microtitre plates; each plate contained 100 μL of parasite culture (0.5% parasitemia, 2.5% hematocrit). Each compound was tested in triplicate and parasite growth compared to control and blank (uninfected erythrocytes) wells. After 24h of incubation at 37°C, 3.7 Bq of [ ³ H]hypoxanthine was added to each well. Cultures were incubated for a further 24 h before they were harvested onto glass-fiber filter mats. The radioactivity was counted using a Wallac Microbeta® 1450 scintillation counter. The results were recorded as counts per minute (CPM) per well at each compound concentration, control and blank wells. Percentage inhibition was calculated from comparison to blank and control wells, and IC ₅₀ values calculated using Microsoft XLFit ® line fitting software (IDBS, UK).

Primary screen The preliminary screen used the 3D7 strain. The compounds were tested at 6 concentrations (30, 10, 3, 1 , 0.3, and 0.1 μg/mL). If the compound did not affect parasite growth at 10 μg/mL it was classified as inactive, between 10 and 1 μg/mL, the compound was designated as partially active, and if <1 μg/mL, the compound was classified as active and was further evaluated by three-fold serial dilutions in a repeat test.

Secondary screen

Both the 3D7 clone and the K1 line were used. The compounds were diluted three-fold over at 12 different concentrations with an appropriate starting concentration based on the preliminary screen. The IC ₅₀ was determined by a sigmoidal dose response analysis using Microsoft XLFit™ (IDBS, UK). For each assay, the IC ₅ o and IC ₉₀ values for each parasite line were determined against the known anti-malarials chloroquine and

artesunate, plus other standard compounds appropriate for the assay. Activity of Standards

IC 50 IC ₅₀

Chloroquine 3D7 4 - - 1 ng/mL K1 130 - - 210 ng/mL Artesunate 3D7 8 - - 2 ng/mL K1 8 - 2 ng/mL 2) Results:

Results obtained for each the tested compounds are showed in Table 1 presented in Example 65 below. EXAMPLE 65: In vitro demonstration of the antiparasitic activity of compounds of formula (1) 1 ) Material and methods:

Plasmodium cultures

P. falciparum GHA strain, derived from a Ghanese patient and chloroquine sensitive have been maintained in RPMI-1640 medium supplemented with 0.37 mM hypoxanthine, 25 mM Hepes, 25 mM NaHCO ₃ , and 10% O ⁺ human serum together with 2-4% washed human O ⁺ erythrocytes. All cultures and assays have been conducted at 37°C under an atmosphere of 4% CO ₂ , 3% O ₂ and 93% N ₂ . Drug sensitivity assays Compound stock solutions were prepared in 100% DMSO at

20 mM or mg/mL. The compound were serially pre-dlluted (2-4-fold) in DMSO followed by a further dilution in demineralised water to assure a final in-test DMSO concentration of <1 %.

Assays were performed in sterile 96-well microtiter plates, each well containing 10 μL of the watery compound dilutions together with 190 μL of the malaria parasite inoculum (1% parasitaemia, 2%HCT). Parasite growth was compared to untreated-infected and uninfected-controls. After 72 h incubation at 37 ⁰ C, plates were frozen at -20 ⁰ C. After thawing, 20 μL of each well was transferred into another plate together with 100 μL Malstat™ reagents and 20 μL of a 1/1 mixture of PES (Phenazine ethosulfat, 0.1 mg/mL) and NBT (Nitro Blue Tetrazolium Grade III, 2 mg/mL). The plates were kept out of light for 2 hours and change in colour was measured

spectrophotometrically at 655 nm. The results were expressed as % reduction in parasitaemia compared to control wells and ICso-values were calculated.

Primary screen

The GHA strain was used. Compounds were tested at 5 concentrations. Artemether and chloroquine were included as reference drugs.

Secondary screen

IC50 values were determinated using an extended dose range (2-fold compound dilutions). Advanced selectivity evaluation was performed against a panel of unrelated organisms (bacteria, yeasts, fungi and other protozoan parasites). 2) Results:

Results obtained for each the tested compounds are presented in Table 1 below: TABLE 1

TABLE 1 (Continued)

TABLE 1 (Continued)

These results show that all the tested compounds of formula (I) have an antiparasitic activity against P. falciparum.

EXAMPLE 66: In vivo demonstration of the antiparasitic activity of compounds of formula (I) 1) Material and methods:

Parasite and animal strains

Plasmodium strains used:

P. berghei ANKA strain, which is chloroquine sensitive (Killick-

Kendrick, 1978).

Female mice (TFW or BALB/c, specific pathogen free), 18-20 g, or

P. chabaudi

General experimental procedure

Infection: Blood was taken from donor mice and diluted in

RPMI-1640 medium to a parasitemia of 1 % (the equivalent of 1x10 ⁷ infected erythrocytes/mL) whilst kept on ice. An inoculum of 0.2 mL was administered to each mouse by the intravenous route. At 2-4 hours post-inoculation dosing commences.

Evaluation endpoint: On day 5 blood smears of all animals (2 slides/mouse) were prepared, fixed with methanol and stained with 10% Giemsa's stain. Parasitemia was determined microscopically by counting 1000 red blood cells under oil immersion at 1000x magnification. Results were reported as % infected erythrocytes and compared to the chloroquine control

group and the untreated control group.

Primary screen

The chloroquine sensitive Plasmodium berghei ANKA strain was used in the primary evaluation. Treatment regimen:

Each of the tested compounds of formula (I) according to the invention was prepared as a stock solution of 5 mg/mL in 10% DMSO/PBS.

Insoluble compounds of formula (I) were milled with stainless steel balls for up to 24 hours to reduce particle size. Fresh dilutions were prepared each day. The route of administration of tested compounds was intra peritoneal (i.p.).

Compounds of formula (I) were given in a 0.2 ml bolus every day for 4 days at

50 mg/kg, i.p. unless in vitro toxicity indicates a narrow therapeutic index and a lower dose was required. The control drug, chloroquine was administered per os (p.o.) every day for 4 days at 10 mg/kg, giving >90% parasite clearance on day 4.

Activity criteria:

Inactive: parasitemia < 25% reduction

Moderate activity: parasitemia 25-75% reduction

Active: parasitemia > 75% reduction Secondary screen

The chloroquine sensitive Plasmodium berghei ANKA strain was used as in the primary evaluation.

Treatment regimen:

Compounds considered active were repeated over a dose range to determine ED ₅₀ ZED ₉₀ values. Compounds were formulated at 10 mg/kg or at 50 mg/kg in a standard suspension vehicle (0.5% carboxymethylcellulose, 0.5% benzyl alcohol, 0.4% Tween® 80 in 0.9% NaCI) for both oral and i.p. administration.

Dosing was over a 4 day period as above, and evaluation was also as above.

Activity Criteria:

Depends upon compound class and therefore control drug

used.

2) Results:

The corresponding results are tabulated in Table 2 below: TABLE 2

EXAMPLE 67: In vitro demonstration of the antitrypanosomal activity of compounds of formula (I)

1 ) Material and methods:

Trypanosoma cultures Trypanosoma brucei rhodesiense STIB 900, a clone of a population isolated in 1982 from a patient in Tanzania which is known to be susceptible to all currently used drugs was used in the study. The bloodstream form trypomastigotes were maintained in MEM medium with Earle's salts supplemented with 25 mM HEPES, 1 g/L additional glucose, 10 mL/L MEM non-essential aminoacids (100x), 0.2 mM 2-mercaptoethanol, 2 mM Na- pyruvate, 0.1 mM hypoxanthine, 0.05 mM bathocuprionedisulphonic acid, 0.15 mM L-cysteine and 15% heat inactivated pooled human serum. All cultures and assays were conducted at 37°C under an atmosphere of 5% CO ₂ /95% air mixture. Drug sensitivity assays

Stock drug solutions were prepared in 100% DMSO (dimethylsulfoxide) at 20 mg/mL, and ball milled or sonicated if necessary. The stocks were kept at 4°C. For the assays, the compound was further diluted to the appropriate concentration using complete medium. Assays were performed in sterile 96-well microtiter plates,

each well containing 100 μL of parasite culture (1 x 10 ³ bloodstream forms) with or without serial drug dilutions at 37°C for 72 hours in 5% CO ₂ . The highest concentration for the test compounds was 30 μg/mL. Each drug was tested in triplicate. A 3-fold serial dilution was performed down to a suitable concentration to obtain an IC ₅₀ value. Initial testing was 30, 10, 3 and 0.1 μg/mL. Control drug was Pentamidine and was diluted down to 0.0001 μg/mL (12 dilutions). Control wells were without drug, blanks were medium only. After 72hours of incubation the plates were inspected under an inverted microscope to assure growth of the controls and to determine the minimum inhibitory concentration (MIC): this is the lowest drug concentration at which no trypanosomes with normal morphology and motility as compared to the control wells can be seen. 20 μL of Alamar Blue were added to each well and the plates incubated for another 2-4 hours. Then the plates werewere read on a Gemini Plate Reader (Molecular Devices) using an excitation wave length of 530 nm and an emission wave length of 580 nm (cut off 550nm).

Primary screen

The preliminary screen used the Trypanosoma b. rhodesiense strain. The compounds were tested at 7 concentrations (drug concentration range from 30 μg/mL to 0.1 μg/mL in 3-fold dilutions). In this assay pentamidine had an ED ₅₀ value of 0.1 to 0.02 ng/mL.

Secondary screen

Active compounds (IC50 <0.2μg/mL) were tested again over the appropriate dose range against Trypanosoma brucei rhodesiense STIB 900 to confirm IC ₅₀ and IC ₉₀ values in comparison to standard pentamidine and internal chemical group standard if available. 2) Results:

Results obtained for each the tested compounds are presented in Table 3 below:

TABLE 3

These results show that all the tested compounds of formula have an activity against T. brucei rhodesiense.