Title of Invention

NOVEL S,S'-[DISULFANEDIYLBIS(3-DISUBSTITUTED AMINO-PROPANE-2, 1-DIYL)]BIS- (DISUBSTITUTERDTHIOCARBAMATE) AND THE PROCESS FOR PREPARATION THEREOF.

Abstract Novel S,S"-[disulfanediylbis(3-disubstituted amino-propane-2, l-diyl)]bis-(disubstitutedthiocarbamate) of general formula (I) as shown in Fig.l of the accompanying drawings , wherein NR1R2 = hetrocyclic amines such as N-methylpiperazine, piperidine, pyrrolidine or morpholine, dialkyl amine such as dimethylamine, diethylamine, methylethylamine, arylatkyl amine such as N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-ethylaniline, or their substituted products, and NR3R4 = hetrocyclic amines such as N-methylpiperazine, piperidine, pyrrolidine or morpholine, dialkyl amine such as dimethylamine, diethylamine, methylethylamine, arylalkyl amine such as N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-ethylaniline, or their substituted products.
Full Text Field of invention
The present invention relates to novel S, S'-[disulfanediylbis(3-disubstituted amino-propane-2, 1-diyl)]bis-(disubstituterdthiocarbamate) and the process for preparation thereof. The invention also provides methods for the preparation of the said spermicidal agents and pharmaceutical composition containing the said compounds. These compounds are useful as spermicidal agents and for preventing unwanted pregnancies as a barrier contraceptive. In addition, the compounds may be useful in the prevention of sexually transmitted microbial infections.
Background of the invention
There exists a constant need for preparing new spermicidal agents with microbicidal activity, which can be utilized as a barrier contraceptive for preventing pregnancies. There has been tremendous emphasis on the development of innovative strategies for the synthesis of spermicidal agents with antimicrobial activity.
Population control is one of the major problems in the world. Among the nonsurgical methods of barrier contraception, the use of spermicides is gaining importance in the recent years. The currently used barrier contraceptives like vaginal contraceptive creams, aerosols, foam tablets contain spermicidal agents like nonyl phenoxy polyethoxy ethanol (commonly known as nonoxynol or nonyl-9), isooctyl phenoxy polyethoxy ethanol , menfegol, sodium lauryl sulfate, phenyl mercuric acetate, quinine etc., but they provide poor pregnancy protection, since they have either poor cervical mucus bio-diffusion ability or poor and unstable foam formation or they cause vaginal irritation. Further, the delivery systems for these spermicidal agents are cumbersome, messy in use and therefore, have poor consumer acceptance. Other known spermicidal agents such as gossypol, thymol, formylated phenols, substituted amino ethyl ketones and saponins are known to be associated with several drawbacks. Therefore, there is a need to develop new molecules, which are safe, efficient, have good cervical mucus bio-diffusion ability, possess spermicidal efficacy with antimicrobial activity and can be conveniently delivered like a stable foam tablet, pessary or coated in condom Prior art references:
It has been reported that sodium salts of dialkyl dithiocarbamate exhibit various biological activity 1-Dialkylaminocarbodithioic acid sodium/potassium salts and their esters have been reported to exhibit antibacterial and antifungal activity by several workers (Ates, Oznur, Kocabalkanli, Ayse, Cesur Nesrin, Otuk, Gulten Farmaco, 1998 53, 542-544, Nofal, Z M , Fahmy, H H , Mohamed, H S Archives of Pharmacol Research, 2002, 25, 28-38, Singh, Nanhai, Gupta, Sushmita, Nath, Gopal Appl Organomet Chem 2000, 14, 484-492, Nakagawa, Susumu, Otake, Nonkayu, Kiyonaga, Hideo, Yamada, Koji, Hona, Hideki, Ushijima,Ryosuke 2001, U S Patent US 6,251,891 1 -Pyrrplidinecarbodithioic acid have been shown to produce antioxidant induced changes in AP-1 transcription complex thereby selectively suppressing the human papillomavirus transcription (Roesl, Frank, Das, Bhudev C , Lengert, Maikei, Geletneky, Kersten, Zur Hausen, Harald , J Virol 1997, 71, 362-370) and it also inhibits the bone resorption (Ozaki, Ken, Takeda, Hiroyuki, Iwahashi, Hiroyoshi Kitano, Shigeo, Hanazawa, Shigemasa FEBS Lett 1997, 410, 297-300) The dithiocarbamates also possess immuno suppressant activity (Martinez-Martinez, Sara, del Arco, Pablo Gomez, Armesilla, Angel Luis, Aramburu, Jose, Luo, Chun, Rao, Anjana, Redondo, Juan Miguel Mol Cell Biol 1997, 17, 6437-6447) and accelerate wound healing (Seitz, Cornelia S , Khavan, Paul A 1997 PCT Int Appl WO 98 44, 908) Antitumor activity has also been reported in dithiocarbamate denvates (Kanekura, T , Higashi, Y , Kenzaki, T Cancer Lett 2000, 161, 177-183) The sodium salt of
pyrrolidinecarbodithioic acid enhanced immunostimulatory effect of interleukin-12 (Trinchien, Geiorgio, Lee, William M F, Kobhsh, Holly, 1998 USA,US 6,375,944) and exhibited antiviral effect on human rhinovirus (Gaudernak, Elizebeth, Seipelt, Joachim, Tnendl, Andrea, Grassauer. Andreas, Kuechler, Ernst J Virology, 2002, 76, 6004-6015)
The diethyl dithiocarbamate induces recruitment and activation or maturation of T-cells by increasing the synthesis of hormone likes factors active on T-cells even in arthymic mice It is also used in the treatment of cancer, immuno-deficiency autoimmune diseases, chronic infections & immunologic impairment due to aging Results from in vitro studies on MIC, MFCs & interaction with anti-fungal agents suggested that dithiocarbamates may be useful adjunct to conventional therapy in the treatment of immuno compromised patients with systemic fungal infections S-(2,6-dialkyl-anilido) carbonyl methyl esters of N, N-disubstituted dithiocarbamic acid was found to show fungicidal and herbicidal activity
In 1959, John et al, (John, H , Holzaepfel, M D, Roy, W , Greonlee, B S , R E Wyant, M S , and Ellis, W C Jr B S , Fertility and sterility 1959,10 , 273-83) reported spermicidal activity of sodium dialkyl dithiocarbamates, specially the dimethyl and diethyl compounds at a test cone of 0 001% and 0 002% respectively while simple alkyl xanthate salts including the amyl compound showed activity at 0 5% cone , since both dithiocarbamate and xanthate salts are unstable under acid conditions Other related compounds derived from the active dithiocarbamates retained only a minor degree of activity In 1983 (Geogiev, VST, Survey of Drug Research in inynunologic Disease, 1983, 1, 403) it has been reported that sodium salt of N, N-diethyl dithiocarbamic acid possesses immunomodulatory and anti-HIV activity In 1992,
(Schreck, R, Meier, B , Mannel, DN Droge, W, Bacunc PA J Exp Med 1992, 175, 1181) it has also been reported that sodium salts of N, N-diethyl dithiocarbamic acid blocks the activation of nuclear factor KB (NFKB) a transcription factor in intact cells It was found that combination of an antioxidant such as dithiocarbamate and an antiviral agent might be useful in the treatment of AIDS patients Diethyl dithiocarbamate also shows antioxidant activity and is a powerful reductant Recently Tnpathi et al (Tripathi R P, Khan, A R, Setty, B S , Bhadun, A P, Acta Pharm, 1996, 46, 169-176) synthesized a number of alkyl/aryl esters of diethyl dithiocarbamates and evaluated them for their spermicidal activity Their structure activity relationship analysis indicated that dithiocarbamate having four to five carbon units with or without hetero atom in the amine part is active Introduction of any additional hydrophobic substitute in the amine part results in loss of spermicidal activity In the 5-alkyl diethyl dithiocarbamate series, the methyl ester retains activity Further, increase in the hydrophobic character in the ester results in the loss of activity However, the introduction of COOH group in the S-alkyl part of the ester (increase in hydrophilicity) retains spermicidal activity Doyle etal [Doyle, W J Jr, 1973 US patent US 3728,371, Chemical Abstrat 1973, 79, 5246a] reported the preparation of dialkylamino carbothioic acid S-(2,3-epithiopropyl) ester by the reaction of epichlorohydnn with sodium salt of dialkylaminodithiocarbamate Thiocarbamates are reported by Davis et al [Davis G E, Driver G W Hoggarth E, Martin A E , Paige MFC, Rose F L and Wilson B R, Brit J Pharmacol 1956, 11, 351-56] as potent antitubercular agents Several workers reported the herbicidal activity of the thiocarbamates Tricyclic carbamate compounds useful for inhibition of farnesyl protein transferase for treatment of cell-proliferative diseases were prepared by Bishop
et al (1998 US patent US 5,721,236) Pesticidal and fungicidal activity in thiocarbamates was reported by Schaper et al ( 1999 Ger Offen DE 19,815,026) Microbicidal activity of S-substituted P-thioacry lam ides was reported by Weinstein et al (1991 Eur Pat Appl EP 440,329) Objects of the invention:
The main object of the invention is to provide novel spermicidal and antifungal agents
Another object of the invention is to provide novel compounds S,S'-[disulfanediylbis(3-disubstituted amino-propane-2,1 -diyl)]bis-(disubstitutedthio-carbamate
Yet another object is to provide pharmaceutical compositions containing the novel compounds of the invention
Still another object is to provide processes for the preparation of the novel compounds of the invention Detailed Description:
The present invention provides S,S'-[disulfanediylbis(3-disubstituted amino-propane-2,l-diyl)]bis-(disubstitutedthiocarbamate) represented by the general formula (1) as shown in Fig 1 of the accompanying drawings, wherein NR1R2 = hetrocyclic amines such as N-methyl piperazine, pipendine, pyrrolidine or morphohne, dialkyl amine such as dimethylamme, diethylamme, methylethylamme, arylalkyl amine such as N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-ethylaniline, or their substituted products and NR3R4 = hetrocyclic amines such as N-Methyl piperazine, pipendine, pyrrolidine or morphohne, dialkyl amine such as dimethylamme,
diethylamine, methylethylamine, arylalkyl amine such as N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-ethylanihne, or their substituted products
Accordingly the present invention provides novel S,S'-[disulfanedrylbis(3-disubstituted amino-propane-2,1 -diyl)]bis-(disubstitutedthiocarbamate) represented by the general formula (1) as shown in Fig 1 of the accompanying drawings, wherein NR1R2 = hetrocyclic amines such as N-methylpiperazme, pipendine, pyrrolidine or morpholme, dialkyl amine such as dimethylamine, diethylamine, methylethylamine, arylalkyl amine such as N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-ethylanihne, or their substituted products and NR3R4 = hetrocyclic amines such as N-methylpiperazme, pipendine, pyrrolidine or morpholme, dialkyl amine such as dimethylamine, diethylamine, methylethylamine, arylalkyl amine such as N-methylanihne, N-methylbenzylamine, N-ethylbenzylamme N-ethylanihne, or their substituted products
The Applicants have developed several compounds using the process discussed herein below The representative compounds (1-14) bearing formulae (II) to (XV) are depicted in figures 1 respectively of the accompanying drawings
(Formula Removed) Formula I
Where:
(Table Removed)
The present invention further provides a process for the preparation of S,S'-[disulfanediylbis-(3-disubstituted amino-propane-2.1-diyl)]bis-(disubstitutedthio-carbamate) compounds having general formula (1) which is shown in Fig 1 of the accompanying drawings, and their salts, said method comprising the steps of
(a) preparing an alkali metal salt of dialkylammo dithiocarbamic acid by conventional methods,
(b) reacting the resulting alkali salt with epihalohydnn at a temperature in the range of -10 to 100° C, for a period in the range of 30 minute to 10 hours, in a protic solvent,
(c) recovering the epithio compound from the reaction mixture by conventional methods, treating the epithio compound with a secondary amine in an appropriate
solvent at a temperature in the range of 0-100° C for a period in the range of 1-100 hours,
(d) recovering the S,S'-[disulfanediylbis(3-disubstituted amino-propane-2,1-diyl)]bis-(disubstitutedthiocarbamate) by conventional methods, and
(e) Converting free base compounds into their salts by conventional methods
In an embodiment the alkali metal salt of the secondary ammo dithiocarbamic acid is prepared by the process reported by R P Tripathi et al, wherein, mixture of secondary amine (0 1 mol) in ether and 30% NaOH (0 1 mol) was treated by drop wise addition of carbon disulfide (0 12) below 5°C The mixture was stirred for one hour, concentrated and recrystalhsed with methanolic ether
In yet another embodiment, the amine is a secondary amine selected from hetrocyclic amines such as N-methyl piperazine, pipendme, pyrrolidine or morpholine, dialkyl amine such as dimethylamine, diethylamine, methylethylamme, arylalkyl amine such as N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-ethylaniline, c their substituted products
In another embodiment, the alkali metal salt of dithiocarbamic acid is selected from lithium, sodium or potassium
In yet another embodiment, the epihalohydrin is selected from epichlorohydnn or epibromohydrin
In another embodiment, the solvents employed, in opening the epithio ring, are selected from the group consisting of ethyl acetate, toluene, benzene, hexane, heptane and chloroform
In yet another embodiment, the amine used, in opening the epithio ring, is a secondary amine selected from hetrocyclic amines such as N-methyl piperazine,
pipendine, pyrrolidine or morphohne, dialkyl amine such as dimethylamme, diethylamine, methylethylamine, arylalkyl amine such as N-methylaniline, N-methylbenzylamme, N-ethylbenzylamme N-ethylanihne, or their substituted products
In a further embodiment, the recovery and purification of the epithio compound is affected by the conventional methods
In yet another embodiment, the preparation of salts from the free base is achieved by reacting free base compound of formula (1) as shown in Fig 1 of the accompanying drawings, with corresponding acids using a lower aliphatic alcohol having C] to C4 carbon atoms as solvent
Further, the water-soluble salts of the free base prepared by using organic or inorganic acids such as hydrochloric acid, citric acid, tartaric acid
The process of the present invention is explained by the reaction scheme depicted in Fig 2 of the accompanying drawings The process consists of the following steps
(a) preparing an alkali metal salt of dialkylammo dithiocarbamic acid of formula (a) by conventional methods,
(b) reacting the resulting alkali salt at a temperature in the range of -10 to 100°C, for a time in the range of 30 minute to 10 hour, with epihalohydrin in a protic solvent,
(c) Extracting the resulting epithio compound with an organic solvent to obtain dialkylammo carbothioic acid S-(2,3-epithio propyl) ester of formula (b), treating the resulting compound with a secondary amine
(d) extracting the S,S'-[disulfanediylbis(3-disubstituted amino-propane-2,1-diyl)]bis-(disubstitutedthiocarbamate), of formula (1), formed with an organic
solvent, followed by recovering and purifying the product as free base of formula (1) as shown in Fig 1 of accompanying drawings, by conventional methods, and
(e) Converting free base compounds into their salts by conventional methods
If desired the salts so formed are used as active ingredients in pessary, foam
tablet, cream, vaginal foam or in medicated condom
Brief Description of the accompanying drawings:
In the accompanying drawings,
Fig 1 represents the general formula of S,S'-[disulfanediylbis(3-disubstituted
aminopropane-2,1 -diyl)]bis-(disubstitutedthiocarbamate)
Fig 2 represents the scheme for the preparation of S,S'-[disulfanediylbis-
(pipendmopropane-2,1 -dryl)]bis-(pyrrolidinothiocarbamate)
Summary of the invention:
Accordingly, the present invention provides novel S,S'-[disulfanediylbis(3-
disubstituted aminopropane-2,1 -diyl)jbis-(disubstitutedthiocarbamate) having
general formula (1) as shown in Fig 1 of the accompanying drawings and useful as
spermicidal agents with antifungal activity
1 S,S'-[disulfanediylbis(pyrrohdino-propane-2,l-diyl)]bis-(pipendinothio-carbamate)
2 S,S'-[disulfanediylbis(pyrrohdino-propane-2,1 -diyl)]bis-(pyrrohdinothio-carbamate)
3 S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,1 -diyl)]bis-(pipendinothiocarbamate)
4 S,S*-[disulfanediylbis(pipendinopropane-2,1 -diyl)]bis-(pipendinothio-carbamate)
5 S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(pyrrohdinothio-carbamate)
6 S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,1 -diyl)]bis-(pyrrohdinothiocarbamate)
7 S,S'-[disulfanediylbis(morpholinopropane-2,1 -diyl)]bis-(piperidinothiocarbamate)
8 S,S'-[disulfanediylbis(morpholinopropane-2,l-diyl)]bis-(methylethyl-aminothiocarbamate)
9 S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(methylethyl-aminothiocarbamate)
10 S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,1 -diyl)]bis-(methy lethy laminoth locarbamate)
11 S,S'-[disulfanediylbis(pipendinopropane-2,1 -diyl)]bis-(morphohnothio-carbamate)
12 S,S'-[disulfanediylbis(N-methylpiperazmopropane-2,l-diyl)]bis-(diethylaminothiocarbamate)
13 S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(diethylamino-thiocarbamate)
14 S,S'-[disulfanediylbis(morphohnopropane-2,l-diyl)]bis-(diethylamino-
th locarbamate)

The invention also provides a method for the preparation of novel S,S'-[disulfanediylbis(3-disubstituted aminopropane-2,1 -diyl)]bis-(disubstitutedthio-carbamate) of formula (I) and their formulations
The advantages of these new compounds as prepared by the process of the present invention are that they provide new compounds useful as spermicidal agents The activity of these compounds is not pH dependent These compounds can be used as vaginal contraceptive agents after incorporation in a suppository, foam tablet, cream, or in a lubricated condom These compounds also inhibit fungal infections
The compounds prepared by the process of the present invention are new and two to ten times more active than nonoxynol-9, a widely used spermicidal agent The spermicidal activity of the compounds prepared by the process of the present invention is not dependent on the pH
These considerations led to explore spermicidal activity in new molecules The applicants synthesized a number of S,S'-[disulfanediylbis(3-disubstituted amino-propane-2,1 -diyl)]bis-(disubstitutedthiocarbamate)
The invention is illustrated by the following examples, which are provided to illustrate the invention and in no way represent a limitation on the inventive concept embodied herein
Example I Preparation of S,S'-[disulfanediylbis(pyrrolidino-propane-2,l-diyl)]bis-(piperidinothiocarbamate) (Compound No 1):
Pipendine (0 1 mol) was taken in diethyl ether (50 ml), to this aqueous NaOH (30%, 0 1 mol) was added by keeping the temperature of reaction mixture below 5°C,
carbon disulfide (0 12 mol) was added drop wise, keeping the temperature below 0°C The resulting mixture was stirred at room temperature for one hour Solvent was distilled off and the crude mass was recrystalhsed by methanolic ether to get pipendino dithiocarbamic acid sodium salt and stored under nitrogen
The finally powdered pipendino dithiocarbamic acid sodium salt (0 1 mol) was dissolved in 60 ml of water and cooled to 0-5° C Epichlorohydnn (0 1 mol) was added with stirring at room temperature for one hour and heated to 80° C for five hours An oily layer was separated out Ethyl acetate was added to this and organic layer was washed with water, dried over anhydrous sodium sulphate and solvent was distilled off to get 1-piperidmocarbothioic acid S-(2,3-epithiopropyl) ester (B)
The 1-pipendinocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol) and pyrrolidine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for 120 hours at room temperature, solvent was distilled off and the resulting viscous mass was purified by using basic alumina column The mass so obtained (64 %) was identified as S.S'-[disulfanediylbis(pyrrolidinopropane-2,1 -diyl)]bis-(pipendinothiocarbamate) (II) of formula (1) through its IR, NMR and mass spectra
S,S'-[disulfanediylbis(pyrrolidinopropane-2,l-diyl)]bis-(pipendinothio-carbamate) (II) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example II Preparation of S,S*-[disulfanediylbis(pyrrolidino-propane-2,l-diyl)]bis-(pyrrolidinothiocarbamate) (Compound No 2):
Pyrrolidine (0 1 mol) was taken in diethyl ether (50 ml), to this aqueous NaOH (30%, 0 1 mol) was added by keeping the temperature of reaction mixture below 5°C, carbon disulfide (0 12 mol) was added drop wise, keeping the temperature below 0°C The resulting mixture was stirred at room temperature for one hour Solvent was distilled off and the crude mass was recrystallised by methanolic ether to get pyrrohdino dithiocarbamic acid sodium salt and stored under nitrogen
The finally powdered pyrrohdino dithiocarbamic acid sodium salt (0 1 mol) was dissolved in 60 ml of water and cooled to 0-5° C Epichlorohydnn (0 1 mol) was added with stirring at room temperature for one hour and heated to 80° C for five hours An oily layer was separated out Ethyl acetate was added to this and organic layer was washed with water, dried over anhydrous sodium sulphate and solvent was distilled off to get 1-pyrrolidinocarbothioic acid S-(2,3-epithiopropyl) ester (B)
The 1-pyrrolldinocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol) and pyrrolidine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for 120 hours at room temperature, solvent was distilled off and the resulting viscous mass was purified by using basic alumina column The mass so obtained (87 %) was identified as S,S'-[disulfanediylbis(pyrrolidinopropane-2,l-diyl)]bis-(pyrrohdinothiocarbamate) (III) of figure (1) through its IR, NMR and mass spectra
S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(pyrrolidinothio-carbamate) (III) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and
stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example III Preparation of S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,l-diyl)]bis-(piperidinothiocarbamate) (Compound No 3).
The 1-pipendinocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol), as prepared in example I, and N-methyl-piperazine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for 120 hours at room temperature, solvent was distilled off and the resulting viscous mass was purified by using basic alumina column The mass so obtained (42%) was identified as S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,l-diyl)]bis-(pipendinothiocarbamate) (IV) of figure (1) through its IR, NMR and mass spectra
S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,1 -dryl)]bis-(pipendinothiocarbamate) (IV) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example IV Preparation of S,S'-[disuIfanedrylbis(pipendinopropane-2,l-dryl)]bis-(pipendinothiocarbamate) (Compound No 4):
The 1-pipendinocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol), as prepared in example I, and pipendine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for 120 hours at room temperature, solvent was distilled off and the
resulting viscous mass was purified by using basic alumina column The mass so obtained (38%) was identified as S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(piperidmothiocarbamate) (V) of figure (1) through its IR, NMR and mass spectra
S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(pipendinothio-carbamate) (V) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example V Preparation of S,S'-[disuIfanediylbis(pipendinopropane-2,l-diyl)]bis-(pyrrolidinothiocarbamate) (Compound No 5):
The 1-pipendinocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol), as prepared in example II, and pipendine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for 120 hours at room temperature, solvent was distilled off and the resulting viscous mass was purified by using basic alumina column The mass so obtained (47 %) was identified as S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(pyrrohdinothiocarbamate) (VI) of figure (1) through its IR, NMR and mass spectra
S,S'-[disulfanediylbis(piperidinopropane-2,l-diyl)]bis-(pyrrohdinothio-carbamate) (VI) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and
stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example VI Preparation of S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,l-diyl)]bis-(pyrrolidinothiocarbamate) (Compound No 6):
The 1-pyrrolidinocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol), as prepared in example I, and pipendine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for 120 hours at room temperature, solvent was distilled off and the resulting viscous mass was purified by using basic alumina column The mass so obtained (60%) was identified as S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,l-diyl)]bis-(pyrrohdinothiocarbamate) (VII) of figure (1) through its IR, NMR and mass spectra
S,S'-[disulfanediy lbis(N-methylpiperazmopropane-2,1 -dryl)]bis-(pyrrohdinothiocarbamate) (VII) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example VII Preparation of S,S'-[disulfanediylbis(morphohnopropane-2,l-diyl)]bis-(piperidmothiocarbamate) (Compound No 7).
The 1-pyrrolidinocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol) was taken in benzene and to this (0 06 mol) of morphohne was added dropwise with stirring, the mixture was heated under reflux for four hours and after the reaction was
complete, as monitored by TLC, solvent was distilled off and the resulting viscous
mass was purified by using silica gel column The mass so obtained (71%) was
identified as S,S'-[disulfanediylbis(morphohnopropane-2,1 -diyl)]bis-
(pipendinothiocarbamate) (VIII) of figure (1) through its IR, NMR and mass spectra
S,S'-[disulfanediylbis(morpholinopropane-2,l-diyl)]bis-(pipendinothio-carbamate) (VIII) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example- VIII Preparation of S,S'-[disulfanediylbis(morphohnopropane-2,l-diyl)]bis-(methylethylaminothiocarbamate) (Compound No 8):
The 1-methylethylaminocarbothioic acid S-(2,3-epithiopropyl) ester (B) and (0 05 mol) of morpholine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for 120 hours at room temperature, solvent was distilled off and the resulting viscous mass was purified by using basic alumina column The mass so obtained (66%) was identified as S,S'-[disulfanediylbis(morphohnopropane-2, l-diyl)]bis-(methylethyl-amino-thiocarbamate) (IX) of figure (1) through its IR, NMR and mass spectra
S,S'-[disulfanediylbis(morpholinopropane-2,l-diyl)]bis-(methylethyl-aminothiocarbamate) (IX) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example IX Preparation of S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(methylethylaminothiocarbamate) (Compound No 9):
The 1-methylethylaminocarbothioic acid S-(2,3-epithiopropyl) ester (B) and
(0 05 mol) of piperidine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for
120 hours at room temperature, solvent was distilled off and the resulting viscous mass
was purified by using basic alumina column The mass so obtained (42%) was
identified as S,S'-[disulfanediylbis(pipendinopropane-2, l-diyl)]bis-
(methylethylamino-thiocarbamate) (X) of figure (1) through its IR, NMR and mass spectra
S ,S'- [disulfanediy lbis(p iper id inopropane-2,1 -diy 1)] bis-(methy lethy lamino-thiocarbamate) (X) (0 05 mol) was dissolved in 50 ml absolute alcohol Citric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The Citrate was precipitated by adding dry diethyl ether
Example X Preparation of S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,l-diyl)]bis-(methylethylaminothiocarbamate) (Compound No 10):
The 1-methylethylammocarbothioic acid S-(2,3-epithiopropyl) ester (B) and (0 05 mol) of N-methylpiperazine (0 05 mol) were dissolved in 75 ml ethyl acetate and stirred for 120 hours at room temperature, solvent was distilled off and the resulting viscous mass was purified by using basic alumina column The mass so obtained (30%) was identified as S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,l-diyl)]bis-

(methylethylaminothiocarbamate) (XI) of figure (I) through its IR NMR and mass spectra
S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,1 -diyl)]bis-(methylethylaminothiocarbamate) (XI) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example XI Preparation of S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(morpholinothiocarbamate) (Compound No 11).
The 1-morpholinocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol) was taken in benzene and to this (0 06 mol) of pipendine was added dropwise with stirring, the mixture was heated under reflux for four hours and after the reaction was complete, as monitored by TLC, solvent was distilled off and the resulting viscous mass was purified by using silica gel column The mass so obtained (42%) was identified as S,S'-[disulfanediylbis(pipendinopropane-2,1 -diyl)]bis-(morphohno-thiocarbamate) (XII) of figure (1) through its IR, NMR and mass spectra
S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(morphohno-thiocarbamate) (XII) (0 05 mol) was dissolved in 50 ml absolute alcohol Tartaric acid (0 1 mol) was dissolved in 25 ml absolute alcohol Both the solutions were mixed and stirred for two hours at room temperature The tartarate was precipitated by adding dry diethyl ether
Example XII Preparation of S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,l-diyl)]bis-(diethylaminothiocarbamate) (Compound No 12):
The 1-diethylaminocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol) was taken in benzene and to this (0 06 mol) of 4-methylpiperazine was added dropwise with stirring, the mixture was heated under reflux for four hours and after the reaction was complete, as monitored by TLC, solvent was distilled off and the resulting viscous mass was purified by using silica gel column The mass so obtained (77%) was identified as S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,1 -diyl)]bis-(diethylammothiocarbamate) (XIII) of figure (1) through its IR, NMR and mass spectra
Example XIII Preparation of S,S'-[disulfanediylbis(pipendinopropane-2,l-diyl)]bis-(diethylaminothiocarbamate) (Compound No 13):
The 1-diethylaminocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol) was taken in benzene and to this (0 06 mol) of pipendine was added dropwise with stirring, the mixture was heated under reflux for four hours and after the reaction was complete, as monitored by TLC, solvent was distilled off and the resulting viscous mass was purified by using silica gel column The mass so obtained (44%) was identified as S,S'-[disulfanediylbis(pipendinopropane-2, l-diyl)]bis-(diethyIammo-thiocarbamate) (XIV) of figure (1) through its IR, NMR and mass spectra
Example XIV Preparation of S,S'-[disulfanediylbis(morpholinopropane-2,l-diyl)]bis-(diethylaminothiocarbamate) (Compound No 14):
The 1-diehtylaminocarbothioic acid S-(2,3-epithiopropyl) ester (B) (0 05 mol) was taken in benzene and to this (0 06 mol) of morpholine was added dropwise with stirring, the mixture was heated under reflux for four hours and after the reaction was complete, as monitored by TLC, solvent was distilled off and the resulting viscous mass was purified by using silica gel column The mass so obtained (47%) was identified as S,S'-[disulfanediylbis(morpholinopropane-2, l-diyl)]bis-(diethyl-aminothiocarbamate) (XV) of figure (1) through its IR, NMR and mass spectra
Example XV In vitro spermicidal activity:
Masturbated semen samples from healthy male volunteers (2 or 3 men) with normal consistency and good quality were liquidated for 30 mm at 37° C Specimens showing good sperm numbers and motility were used in this study
The compounds or nonoxynol-9 (Standard) were dissolved in physiological sahne/DMSO at different concentrations Two drops of semen were placed on a slide and to this two drops of the solution of the compound or nonoxynol-9 were added and the contents mixed (about S seconds) and immediately examined under a phase contrast microscope The results were scored positive if 100% of spermatozoa became instantaneously immobile, whereas if even one or two spermatozoa showed sluggish motility, the test concentration of the compound was scored negative The control slides were prepared by adding two drops of physiological saline The minimum
effective concentration of active compound was determined and repeated in two different sperm samples
The compounds listed in table 1 were prepared and tested for their spermicidal activity against human spermatozoa Some of the compounds showed 100% spermicidal activity at a concentration range 0 5% - 0 01%
TABLE-1 (Table Removed)

EXAMPLE XVI In vivo evaluation of S,S'-[disulfanedivlbis(N-methvlDiDerazinoproDane-2,l-divl)lbis-(DiDendinothiocarbamate) in rabbit model
Procedure: Adult female BeJgian rabbits showing both vaginal and psychic signs of estrus were selected for experimental use Aliquot of pure compound/suppository were prepared in physiological saline before instillation into the rabbit vagina Solutions containing 100 mg (MEC X 100, pure compound) and 50 mg (MEC X 50, in formulation) of S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,1 -diyl)]bis-(pipendinothiocarbamate) was deposited at the cervix of each dose with a syringe attached to an 18 cm long rubber catheter with a 3 mm bore size Within 2-3 minutes following the intravaginal administration, each doe was mated once to a fertile buck
A control (same series) was also run in which the applicator was similarly inserted in the vagina, and the placebo (suppository, dissolved in 1 0 ml saline) was instilled Each dose was tested in at least 4-5 rabbits (William W L Contraception 1980,22, 659)
The rabbits were sacrificed on day 15 of gestation Implantation sites and number of Corpora lutea were recorded
Results: None of the treated females showed implantation sites, though the ovulation was normal as revealed by the number of corpora lutea The control animals showed normal number of implantation sites (4-6)
Example XVII Suppository formulation
1. Preparation of placebo suppository
A mixture of polyethylene glycol (mol wt 1500) 3 0 g, polyethylene glycol (mol wt 4000) 2 0 g, polyethylene glycol (mol wt 6000) 5 0 g and hexane-1-ol (0 6 ml) was mixed and melted by heating on water bath The mixture was allowed to cool to get solid mass, which was moulded, into a suppository of oval shape and 2x1x05-cm size It was then packed to avoid coming into contact with moisture before use
2. Preparation of suppositories of S,S'-[disulfanediyIbis(N-methylpiperazino-
propane-2,l-diyl)]bis-(pipendinothiocarbamate).
A mixture of polyethylene glycol (mol wt 1500) 2 0 g, polyethylene glycol (mol wt 4000) 1 33 g, polyethylene glycol (mol wt 6000) 3 33 g and hexane-1-ol (0 4 ml) was melted by heating on a water bath and 20 mg S,S'-[disulfanediylbis(N-methylpiperazmopropane-2,l-diyl)]bis-(pipendinothiocarbamate) as the spermicidal agent in 2 ml of chloroform was added to it with intimate mixing chloroform was evaporated off by warming the mixture to 60°C The mixture was allowed to cool to get a solid mass, which was moulded, into a suppository of oval shape It was then packed to avoid coming into contact with moisture before use
Example XVIII
Preparation of medicated condoms of S,S'-[disulfanediyIbis(N-methylpiperazinopropane-2,l-diyl)]bis-(pipendinothiocarbamate):
S,S'-[disulfanediylbis(N-methylpiperazinopropane-2,l-diyl)]bis-'
(piperidinothiocarbamate) (20 mg) as the spermicidal agent in 1 ml of polyethylene
glycol was lubricated on the inner tip of the condom It was then packed to avoid coming into contact with moisture before use
Example XIX
In vitro anti fungal activity-
MIC determination: The MIC of compounds were determined by broth microdilution
technique as per guide lines of National Committee for Clinical Laboratory Standards using RPMI 1640 media buffered with MOPS (3-[N-Morphohno] propanesulfonic acid) Starting inoculum of test culture was 1-5x103 CFU/ml Microtiter plates were incubated at 35°C MICs and IC50 were recorded after 48 h of incubation The results of the antifungal activity of some of the compounds are given in table -2
Table- 2
MIC (µg/ml) and MFC(µg/ml) of compounds against different Candida strains:

(Table Removed)
MIC Minimum inhibitory concentration
IC50, concentration at which 50% growth is inhibited
Ca Candida albicans
PK-03 Candida albicans MTCC 183
PK-09 Candida albicans MTCC 1346
PK-13 Candida albicans ATCC 10231 PK-14 Candida krusei ATCC 6258 PK-15 Candida parapsilosis ATCC 22019 PK-17 Candida albicans ATCC 10453 PK-18 Candida albicans ATCC 60193 PK-19 Candida albicans ATCC 66027







We claim
1. Novel S, S'-[disulfanediylbis(3-disubstituted amino-propane-2, 1-diyl)]bis-(disubstituterdthiocarbamate) of general formula (I) as shown in Fig.1 of the accompanying drawing, wherein NR1R2 =hetrocyclic amines selected from N-methylpiperazine, pineridine, pyrrolidine or morpholine, dialkyl amine selected from dimethylamine, diethylamine, methylethylamine, arylalkyl amine selected from N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-ethylaniline and NR3R4 =hetrocyclic amines selected from N-methylpiperazine, pineridine, pyrrolidine or morpholine, dialkyl amine selected from dimethylamine, diethylamine, methylethylamine, arylalkyl amine selected from N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-ethytlaniline and the process for the preparation thereof.
2. The representative compounds of S, S'-[disulfanediylbis(3-disubstituted amino-propane-2, 1-diyl)]bis-(disubstituterdthiocarbamate) comprising :
i) S, S'-[disulfanediylbis (pyrrolidino-propane-2,1diyl)]bis-
piperidinothio-cabamate).
ii) S, S'-[disulfanediylbis (pyrrolidino-propane-2,1diyl)]bis-
piperidinothio-cabamate).
iii) S, S'-[disulfanediylbis (N-methylpiperazinopropane-21-diyl)]
piperidinothio-cabamate).
iv) S, S'-[disulfanediylbis (piperidinopropaane-2,1-diyl)] bis-
piperidinothio-cabamate).
v) S, S'-[disulfanediylbis (piperidinothio-cabamate)2,1diyl)]bis-
pyrrolidinothiocarbamate).
vi) S, S'-[disulfanediylbis (N-methylpiperazinopropane-21-diyl)]
piperidinothio-carbamate).
vii) S, S'-[disulfanediylbis (morpholinopropane-2,1-diyl)] piperidinothio-carbamate).
viii) S, S'-[disulfanediylbis (morpholinopropane-2,1-diyl)] bis -
(methylethyl-aminothiocarbomate)
ix) S, S'-[disulfanediylbis (piperidinopropaane-2,1-diyl)] bis-
(methylethyl-anubitguicarvanate),
x) S, S'-[disulfanediylbis (N-methylpiperazinopropane-21-diyl)] bis-
(methylethylaminocarbamate).
xi) S, S'-[disulfanediylbis (piperidinopropaane-2,1-diyl)] bis-
(morpholinothiocarbamate).
xii) S, S'-[disulfanediylbis (N-methylpiperazinopropane-21-diyl)] bis-
(diethylaminothiocarbamate).
xiii) S, S'-[disulfanediylbis (piperidinopropaane-2,1-diyl)] bis-
(diethylamino-tguicarvanate),
xiv) S, S'-[disulfanediylbis (morpholinopropane-2,1-diyl)] bis
(diethylamino-thiocarbamate).
3. A process for the preparation compounds of S, S'-[disulfanediylbis(3-
disubstituted amino-propane-2, 1 -diyl)]bis-(disubstituterdthiocarbamate)
having general formula (1) as shown in Fig. 1 of the accompanying
drawings, wherein NR1R2 =hetrocyclic amines such as N-
methylpiperazine, pineridine, phrrolidne or morpholine, dialkyl amine such
as dimethylamine, diethylamine, methylethylamine, arylalkyl amine such
as N-methylaniline, N-methylbenzylamine, N-ethylbenzylamine N-
ethytlaniline, or their substituted products, and NR3R4 =hetrocyclic amines
such as N-methylpiperazine, pineridine, phrrolidne or morpholine, dialkyl
amine such as dimethylamine, diethylamine, methylethylamine, arylalkyl
amine such as N-methylaniline, N-methylbenzylamine, N-
ethylbenzylamine N-ethytlaniline, or their substituted products and said
process comprising the steps of:
a) preparing an alkali metal salt of dialkylaminodithocrabamic acid of Formula (a) by conventional methods,
b) reacting the resulting alkali salt with epihalohydrin at a temperature in the range of 10-100 C, for a time in the range of 30 minute to 10 hour in a protic solvent,
c) recovering the epithio compound Formula (b) from the reaction mixture by known methods,
d) treating the epithio compound with an amine at a temperature in the range of 0-100°C for a period in the range of 1-100 hours in a nonprotic solvent,
e) recovering the S, S'-[disulfanediylbis(3-disubstituted amino-propane-2, 1-diyl)]bis-(disubstituterdthiocarbamate) having general formula (1) by conventional methods, and
f) converting free base compounds into their salts by conventional methods.

4. A process as claimed in claim 3 wherein the alkali metal salt of dialkylamino dithiocarbamic acid is selected from lithium, sodium or potassium salts.
5. A process as claimed in claim 3 wherein the epihalohydrin is selected from epichlorohydrin and epibromohydrin.
6. A process as claimed in claim 3 wherein the protic solvent is selected from the group comprising water, ethanol, methanol, benzene, ethanolic or methanolic benzene, toluene, ethanolic or methanolic toluene, chloroform, ethyl acetate or mixtures thereof.
7. A process as claimed in claim 3 wherein the nonprotic solvent is selected from the group comprising ethylacetate, methylpropionate, benzene, toluene, hexane, heptane, chloroform or mixtures thereof.
8. A process as claimed in claim 3 wherein the secondary amine is selected from hetrocyclic amine such as N-methylpiperazine, piperidine, pyrrodine or morpholine, dialkyl amine such as dimethylamine, diethylamine, methylethylamine, arylakyl amine such as N-methylamiline, N-ethylbenzylamine N-ethylaniline, or their substitute dproducts.
9. A process as claimed in claim 3 wherein the epithio compound obtained in
step © is extracted with chloroform, dichloromethane, toluene,
ethylacetate dried and purified by column chromatography.
10. A process as claimed in claim 3 wherein the free base compounds in step
(f) are converted to their salts by reacting free base compound of formula
of figure (1) as claimed in claims 3 and 4 with corresponding acids using a
lower C1 to C4 aliphatic alcohol as solvent.
11. A process as claimed in claim 3 wherein the salts of the free base are
prepared by using organic or inorganic acids such as hydrochloric acid,
citric acid, tartaric acid.
12. Novel S, S'-[disulfanediylbis(3-disubstituted amino-propane-2, 1-diyl)]bis-(disubstituterdthiocarbamate) and the process for preparation thereof substantially as herein described with reference to the examples and drawings accompanying the specification.

Documents:

1792-del-2004-abstract.pdf

1792-DEL-2004-Claims-(18-01-2011).pdf

1792-del-2004-claims.pdf

1792-DEL-2004-Correspondence-Others-(18-01-2011).pdf

1792-del-2004-correspondence-others.pdf

1792-DEL-2004-Description (Complete)-(18-01-2011).pdf

1792-del-2004-description (complete).pdf

1792-del-2004-drawings.pdf

1792-DEL-2004-Form-1-(18-01-2011).pdf

1792-del-2004-form-1.pdf

1792-del-2004-form-18.pdf

1792-DEL-2004-Form-2-(18-01-2011).pdf

1792-del-2004-form-2.pdf

1792-del-2004-form-3.pdf

1792-del-2004-form-5.pdf


Patent Number 245815
Indian Patent Application Number 1792/DEL/2004
PG Journal Number 05/2011
Publication Date 04-Feb-2011
Grant Date 02-Feb-2011
Date of Filing 22-Sep-2004
Name of Patentee COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH
Applicant Address RAFI MARG NEW DELHI-110 001, INDIA.
Inventors:
# Inventor's Name Inventor's Address
1 ANIL KUMAR DWIVEDI CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
2 VISHNULAL SHARMA CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
3 NIHARIKA KUMARIA CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
4 RAJA ROY CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
5 RAM CHANDRA GUPTA CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
6 RAGHWEDNRA PAL CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
7 KIRAN KUMAR CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
8 GOPAL GUPTA CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
9 JAGDAMBA PRTASAD MAIKHURI CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
10 JANAK DULARI DHAR CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
11 PRADEEP KUMAR CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
12 ABDUL HAQ ANSARI CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
13 PRAVEEN KUMAR SHUKLA CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
14 MANISH KUAMR CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
15 KUNNATH PADMANABHAN MADHUSUDANAN CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
16 PRATIMA SRIVASTAVA CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
17 SATYAWAN SINGH CENTRAL DRUG RESEARCH INSTITUTE, CHATTAR MANZIL PLACE, LUCKNOW-226 001 (U.P.), INDIA.
PCT International Classification Number A01N 47/10
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA