WO2009110002A1 - Novel efflux pump inhibitors - Google Patents

Novel efflux pump inhibitors Download PDF

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Publication number
WO2009110002A1
WO2009110002A1 PCT/IN2009/000144 IN2009000144W WO2009110002A1 WO 2009110002 A1 WO2009110002 A1 WO 2009110002A1 IN 2009000144 W IN2009000144 W IN 2009000144W WO 2009110002 A1 WO2009110002 A1 WO 2009110002A1
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formula
acrylic acid
compound
dihydronapth
substituted
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PCT/IN2009/000144
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French (fr)
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WO2009110002A4 (en
Inventor
Surrinder Koul
Niranjan Thota
Venkat Reddy Mallepally
Payare Lal Sangwan
Subhash Chandra Taneja
Inshad Ali Khan
Ashwani Kumar
Alsaba Fida Raja
Ajit Kumar Saxena
Satyam Kumar Agrawal
Rakesh Kamal Johri
Sheikh Tasduq Abdullah
Gurdarshan Singh
Lakshmi Narasamma Rao Bachu
Autar Krishan Dhar
Basant Purnima
Ghulam Nabi Qazi
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Council Of Scientific & Industrial Research
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Publication of WO2009110002A1 publication Critical patent/WO2009110002A1/en
Publication of WO2009110002A4 publication Critical patent/WO2009110002A4/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/02—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having nitrogen atoms of carboxamide groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals
    • C07C233/11—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having nitrogen atoms of carboxamide groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals with carbon atoms of carboxamide groups bound to carbon atoms of an unsaturated carbon skeleton containing six-membered aromatic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/12—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by halogen atoms or by nitro or nitroso groups
    • C07C233/15—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by halogen atoms or by nitro or nitroso groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/16—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
    • C07C233/24—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring
    • C07C233/29—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring having the carbon atom of the carboxamide group bound to an acyclic carbon atom of a carbon skeleton containing six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/45—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C07C233/53—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring
    • C07C233/55—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring having the carbon atom of the carboxamide group bound to a carbon atom of an unsaturated carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/32—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton containing six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D207/06—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with radicals, containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D211/40—Oxygen atoms
    • C07D211/44—Oxygen atoms attached in position 4
    • C07D211/46—Oxygen atoms attached in position 4 having a hydrogen atom as the second substituent in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C07D233/60—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms with hydrocarbon radicals, substituted by oxygen or sulfur atoms, attached to ring nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
    • C07D295/182—Radicals derived from carboxylic acids
    • C07D295/185—Radicals derived from carboxylic acids from aliphatic carboxylic acids

Definitions

  • the present invention relates to the field of chemotherapeutics, particularly use of novel synthetic aromatic amides for potentiating the bio-efficacy of specific drugs.
  • the invention particularly relates to the preparation and use of synthetic analogues of aromatic amides, useful in potentiating bio-efficacy of anti-infective drugs.
  • the invention more particularly relates to aromatic substituted acrylic acid amide of general formula 1 including its analogues or and salts thereof
  • novel amide when administered in combination with an anti-infective drag potentiates the bioactivity of the drug thereby requiring lower doses and/or decreased frequency of dosing while maintaining the therapeutic efficacy of standard doses of such drugs.
  • the synthetic molecules of this invention are all novel, as they have not been reported 5 earlier in the literature. These novel molecules possess specific properties of potentiating the bio-efficacy of specific drugs particularly they are useful in combination with anti-infective drugs which besides reducing their effective dosages are also associated with lesser side effects.
  • the molecules along with the pharmaceutical products/combinations possessing the properties disclosed in the present invention are novel and not known in literature or prior LO art.
  • Pumps may be specific for one substrate or may transport a range of structurally dissimilar compounds (including antibiotics of multiple classes); such pumps can be associated with multiple drug resistance (MDR).
  • MDR drug resistance
  • efflux transporter O. 5 lom ⁇ vskaya, M.. S. Warren. A; Lee, J. Galazzo, R 1 fronko, M. Lee, J, Blais, D. Cho, S. Chamberland, T.ranau, R. Leger, S. Hecker, W. Watldns, K. oshino, H. ishida, V. J. Lee., Antimicrob. Agents and Chemother.
  • MF major facilitator
  • MATE multidrug and toxic efflux
  • RND resistance-nodulation-division
  • SMR small multidrug resistance
  • ABC ATP binding cassette
  • MDR is mainly conferred by MFS efflux systems, the most studied pumps being Nor A of S. aureus (A. A. Neyfakh, C.M.Borsch, G.W.Kaatz Antimicrob. Agents and Chemotherapy, 1993, 37: 128-129) and its homologues in B.
  • the main object of the present invention is to provide the novel 3-(l-substituted-3,4- dihydro-naphth-2-yl)-acrylic acid amide, its tetra hydro derivatives of general formula Ia, 10 laa and 5-(l-substituted-3,4-dihydronaphth-2-yl)-4-alkyl-2E,4E-pentadienoic acid amides its tetrahydro derivatives of structure formulae Ib, lba its analogues and /or their salts thereof.
  • Another object of the invention is to provide the novel aromatic substituted pentadienoic acid amide, which may be useful as potentiators of the bioefficacy of the drugs. 15 Yet another object of the invention is to provide the process for the preparation of the novel aromatic amides.
  • Still another objective of the invention is to provide the compounds of formula 1, which are not toxic.
  • Further object of the invention is to provide the pharmaceutical composition using effective ZO amount of one or more compound of formula Ia including its geometrical isomers, its analogues or and salts thereof as stated above along with the anti-infective drug and optionally along with a carrier or diluent or pharmaceutically acceptable exciepient.
  • Another object of the invention is to provide the pharmaceutical composition, which is useful for the treatment of the infections caused by bacteria. .5 Further object of the invention is to provide the pharmaceutical composition which is used in the reduction in the dose requirement of anti-infectives.
  • the present invention provides a new class of bacterial efflux pump inhibitors(EPIs) (not »0 reported in the literature) for therapeutic use when co-administered with antiinfectives (antibiotics).
  • EPIs bacterial efflux pump inhibitors
  • the EPIs as per the present invention belongs to the general formula 1
  • Staphylococcus aureus 1199 and its mutant Nor A over expressing Staphylococcus aureus 1199B of the genus Gram +ve bacteria known for its resistance to a host of antibiotics (including Ciprofloxacin) when treated with a formulation comprising of antibiotic ciprofloxacin and an EPI (selected from the compounds of the formula 1) in a given definite ratio by potentiation of the drug activity which is because of the inhibition of the efflux pump present in the bacteria.
  • the present invention describes a new class of bacterial efflux pump inhibitors (EPIs) for therapeutic use when given in combination with antibiotics.
  • EPIs bacterial efflux pump inhibitors
  • the EPIs as per the present invention are selected from the general formula 1 which contains compounds that belong to 3-(l-substituted-3,4-dihydro-naphth-2-yl)-acrylic acid amide, its tetra hydro derivatives (general formula Ia, laa) and 5-(l-substituted-3,4-dihydronaphth-2-yl)-4-alkyl-2E,4E- pentadienoic acid amides its tetrahydro derivatives (general formulae Ib, lba) its analogues and /or their salts thereof.
  • a formulation comprising of an antibiotic such as ciprofloxacin is co-administered with an EPI selected from the above mentioned class of compounds and tested against drug resistance bacterial strains such as Staphyllococcus aureus 1199 and its mutant Nor A over expressing Staphyllococcus aureus 1199B (Gram +ve bacteria known for its resistance to aJiost of antibiotics including Ciprofloxacin,mupirocin ).
  • the drug potency is restored due to the inhibition of the bacterial efflux pump as evidenced by the experiments using ethidium bromide (EtBr) where the EPI inhibits the fast loss of fluorescence of EtBr which in absence of EPI is depleted fast.
  • EPIs bacterial efflux pump inhibitors
  • the EPIs as per the present invention are selected from the general formula 1 which contains compounds that belong to 3-(l-substituted-3,4-dihydro-naphth-2-yl)-acrylic acid amide, its tetra hydro derivatives (general formula Ia, laa) and 5-(l-substituted-3,4- dihydronaphth-2-yl)-4-alkyl-2£,4ii-pentadienoic acid amides its tetrahydro derivatives (general formulae Ib, lba) its analogues and /or their salts thereof.
  • general formula 1 which contains compounds that belong to 3-(l-substituted-3,4-dihydro-naphth-2-yl)-acrylic acid amide, its tetra hydro derivatives (general formula Ia, laa) and 5-(l-substituted-3,4- dihydronaphth-2
  • a formulation comprising of an antibiotic such as ciprofloxacin is co-administered with an EPI selected from the above mentioned class of compounds and tested against drug resistance bacterial strains such as Staphyllococcus aureus 1199 and its mutant Nor A overexpressing Staphyllococcus aureus 1199B (Gram +ve bacteria known for its resistance to a host of antibiotics including Ciprofloxacin,mupirocin ).
  • Figure-IB depicts the inhibition of efflux of ethidium bromide from the wild and mutant strain of S. aureus when pretreated with the compound of formula Ia, resulting in no loss of fluorescence compared to controls where the compound of formula I is not present, which leads to rapid decrease in the fluorescence.
  • the present invention provides a substituted aromatic amides of general formula Ia, Ib, laa, lba and salts thereof as potentiators of bioefficacy of anti-infective drugs when used in combination with anti-infective drugs in the form of a pharmaceutical composition.
  • R 5 represents hydrogen atom or normal or branched chain Ci to Cio alkyl group or phenyl or benzyl radical and R 6 represents hydrogen atom or normal or branched chain C 1 to C 10 alkyl group or phenyl or benzyl radical; where NR 5 R 6 together (R 5 H-R 6 ) represent heterocyclic amine radical " such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N- methylpiperazinyl, oxazolyl, N-methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like and optionally converting them to their salts by method icnown in the art of synthesis.
  • R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO), aryl, benzyl (unsubstituted and substituted);
  • R 5 represents hydrogen atom or normal or branched chain Ci to C 10 alkyl group or phenyl or benzyl radical and R 6 represents hydrogen atom or normal or branched chain C 1 to Cjo alkyl group or phenyl or benzyl radical; where NR 5 R 6 together (R 3 -+R 6 ) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N- methylpiperazinyl, oxazolyl, N-methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like.
  • NR 5 R 6 together (R 3 -+R 6 ) represent heterocyclic amine radical such as piperidiny
  • R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO), aryl , benzyl (unsubstituted and substituted);
  • R7 represents normal or branched chain Cl to ClO alkyl group.
  • the salt is pharmaceutically acceptable selected from hydrochloride, acetate, succinate, maleate.
  • the present invention provides a process for the preparation of substituted aromatic acid amides of general formula Ia, laa, Ib and lba as stated above, the method of preparation comprising;
  • R is selected from a group consisting of halogen, hydrogen, alkyl (Ci to Ci 0 ) or aryl or benzyl (unsubstituted and substituted);
  • Ri,R2,R 3 ,R 4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, substituted pyranyl radical, unsubstituted pyi'anyl radical or
  • Ri 1 R 4 represents hydrogen group and R2+R3 together represent 1,3-dioxol or 1,4-dioxol to Wittig reaction (Wittig reagent prepared from triphenylphosphine and ethyl bromoacetate/ ethylchloroacetate
  • Formula 5 (viii) condensation of the compounds of the formula 5 with an ylide (Wittig reagent, prepared from triphenylphosphine and ethyl bromoacetate/ ethylchloroacetate) in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at a temperature ranging between 5-80 0 C for a period ranging between 1 to 24 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like, or benzene/toluene or dimethylformamide followed by direct saponification of Wittig reaction product furnished compounds of the formula 6 wherein R 11 R 2 R 31 R 4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, allceny ⁇ oxy, substituted pyranyl radical, unsubstituted pyranyl radical or Ri 1 R 4 represents hydrogen group and R 2 ,R 3 represent hydrogen, alkoxyl (C 1
  • Formula 6 (ix) treating the compound of the general formula 6 with thionyl chloride in presence of a base such as pyridine followed by treatment of the acid chloride with an appropriate base afforded compounds of the formula Ib wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO) or aryl or benzyl (unsubstituted and substituted); R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, sxibstituted pyranyl radical, unsubstituted pyranyl radical or Rl, R4 represents hydrogen group and R2, R3 represent hydrogen, alkoxyl (Cl-ClO), alkenoxyl groups (Cl-ClO) 1 , aryloxyl, substituted aryloxyl groups or Rl, R4 represents hydrogen group and R2+R3 together represents 1,3-dioxol or 1,4-d
  • the process further comprises converting the compound of formula Ia,laa4b and lba into a salt.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising an effective amount of one or more compound of formula la,laa,lb and lba its analogues or /and salts thereof as stated above along with the anti-infective drug and optionally along with a carrier or diluent or pharmaceutically acceptable exciepient.
  • the amount of compound of formula 1 may vary from 0.1 to 50% by weight of the composition with respect to the drug. In another embodiment of the invention wherein the composition is useful as an anti - microbial agent.
  • composition is reduced more than 32 times of the anti- infective drugs when used alone.
  • ED50 of anti-infective drug reduced to about 1/4 when used in combination with the compound of formula Ia, laa, Ib and lba.
  • the anti-infectives are selected from groups comprising of penicillins (including semi-synthetic), cephalosporins, aminoglycosides, glycopeptides, fluroquinolones, macrolides, tetracyclines other antibiotic groups such as mupirocin and framycetin, first and second line anti-TB drugs, anti-leprosy ' drugs, oxazolidinones.
  • composition is effective against micro- organisms selected from gram positive bacteria and gram negative bacteria, that include
  • Staphylococcus species Bacillus species. Pseiidomonas species, E coli and Salmonella species as well as against micro-organisms such as Mycobacterium species.
  • composition is effective against growth of micro-organisms that include anti-infective resistant strains such as MRSA.
  • composition is effective when tested for curing of mice, guinea pig or rabbit models infected with micro-organisms from the gram positive group of bacteria and gram negative group of bacteria such as Staphylococcus species,
  • compositions including Bacillus species., Pseiidomonas species, E coli, Salmonella species besides being effective against microorganisms such as Mycobacterium species.
  • excipient or pharmaceutical vehicle such as lactose, corn starch, polyethylene glycols and the like may be administered through oral route.
  • compositions along with excipient or pharmaceutical vehicle such as castor oil, olive oil and the like may be administered through systemic route.
  • excipient or pharmaceutical vehicle such as castor oil, olive oil and the like
  • compositions along with excipient or pharmaceutical vehicle such as polyethylene glycols, bees wax, paraffin wax, emulsifying agents and the like may be applied topically.
  • excipient or pharmaceutical vehicle such as polyethylene glycols, bees wax, paraffin wax, emulsifying agents and the like
  • a method for inhibiting a bacterial cell that employs an efflux pump resistance mechanism comprising contacting the cell with a pharmaceutical composition comprising of an antibacterial agent and a compound of formula la,laa,lb,lba and optionally an excipient or pharmaceutical vehicle.
  • the bacterial cell is selected from gram positive bacteria such as Staphylococcus species and Bacillus species.
  • the bacterial cell is selected from gram negative bacteria such as Pseudomonas species, E coli and Salmonella species.
  • the bacterial cell is selected from mycobacterial species.
  • a method of treating infections comprising administrating to a subject in need of such treatment a therapeutically effective amount of compound of formula Ia, laa, Ib, lba, its analogues or/ and salts thereof as claimed in claim 1 along with the anti-infective drug and optionally along with a carrier or diluent or pharmaceutically acceptable exciepient.
  • the aromatic amide of formula 1 i.e. 3-(l-substituted-3,4-dihydronaphth-2-yl)-acrylic acid amide, including its related tetralin derivatives of structural formulae Ia and 5-(l- substituted-3,4-dihydronaphth-2-yl)-4-alkyl-2E,4E-pentadienoic acid amide of formula Ib including its tetralin derivatives of structure formulae lba where R and R 1 to R 7 are described as above, were synthesised from the corresponding Tetralone and eugenol and their derivatives by the known state of art as described in literature.
  • Alpha.-tetralone (1,2,3,4-tetrahydronaphtha-l-one) was first reacted with Vilsmeier reagent comprising of dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tribromide mixture at 0-10 0 C for 5-12 hrs, contents were neutralised with dilute alkali solution to produce l-halo-2-formyl-3,4-dihydronaphthalene.
  • the l-halo-2-formyl- 3,4-dihydronaphthalene was condensed with an ylide (Wittig reagent, prepared from triphenylphosphine and ethyl bromoacetate/ethylchloroacetate) in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at temperature 5 - 80 0 C for 1 to 24 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like benzene/toluene or dimethylformamide to yield corresponding 3-(l-halo-3,4-dihydro-naphth-2-yl)-acrylic acid ethyl ester which was hydrolysed with a strong alkali solution using sodium hydroxide or potassium hydroxide followed by acidification to furnish 3-(l-halo-3,4-dihydronaphth-2-yl) acrylic acid.
  • a solution of aryl alkenoic acid as prepared above in an inert organic solvent such as benzene, dichloromethane was treated with thionyl chloride and excess of solvent removed in vacuo.
  • the acyl chloride intermediate thus obtained without purification was condensed with acyclic or cyclic or heterocyclic amine in inert organic solvent such as dichloromethane, benzene, diethyl ether and the like in the temperature range of 5-50 0 C, and after the purification by crystallisation or column chromatography to produce amide of formula 1.
  • the acid of formula 1 was converted to its tetrahydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and at 1-3 atmospheric pressure to obtain corresponding tetrahydro derivative which was converted to corresponding amides of formula Ia by the method described as above for the preparation of compound of formula 1.
  • polar solvent such as methanol or ethanol
  • Ia may also be prepared by hydrogenation of compound of formula 1 as such by the method as described above. .
  • Method-3 The tetralone was first reacted with dimethylformamide and phosphorus oxybromide or phosphorus tri bromide mixture at 0-10 0 C for 5-24 hrs, contents neutralised with dilute alkali solution to produce l-halo-3,4-dihydronaphthalene-2-carbaldehyde .
  • alkyl magnesium halide alkyl halide C 2 -C 1 O
  • an anhydrous ether solvent such as diethyl ether, tetrahydrofuran and the like to produce corresponding l-(l-halo-3,4- dihydronaphth-2-yl)-alkan-l-ol which was treated with dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tribromide mixture at 0-10 0 C for 20-60 hrs, contents neutralised with dilute alkali solution to produce 3-(l-halo-3,4-dihydronaphth- 2-yl)-2-alkyl-propenal.
  • the 3-(l-halo-3,4-dihydronaphth-2-yl)-2-alkyl-propenal was condensed with an ylide (Wittig reagent, prepared from triphenylphosphine and methyl/ethyl bromoacetate or methyl/ethyl chloroacetate in equimolar.
  • ylide Wood reagent, prepared from triphenylphosphine and methyl/ethyl bromoacetate or methyl/ethyl chloroacetate in equimolar.
  • the acid of formula Ib was converted to its tetrohydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and 1-5 atmospheric pressure to obtain corresponding tetrohydro derivative which was converted to corresponding amides of formula lba by the method described above for the preparation of compound of formula Ib.
  • polar solvent such as methanol or ethanol
  • lba may also be prepared by hydrogenation of compound of formula Ib as such by the method as described above. And excess of thionyl chloride and solvent removed.
  • the acid chloride intermediate thus obtained was condensed with acyclic or cyclic or heterocyclic amine or aryl amine or C-protected amino ester or unsubstituted and substituted allcylaryl amines.
  • the ⁇ -tetralone was first converted to 1,2,3,4-tetrahydronapth-l-ol by reduction with sodium borohydride and the resulting alcohol reacted with dimethylformamide and phosphorus oxybromide or phosphorus tribromide mixture at 0-10 0 C for 5-24 hrs, contents neutralised with dilute alkali solution to produce 3,4-dihydronaphthalene-2-carbaldehyde.
  • the 3,4-dihydronaphthalene-2-carbaldehyde was reacted with malonic acid in presence of base such as piperidine using pyridine as solvent, and the reaction left at room temperature for 24-30 hrs after that heated at 80-90 0 C, contents were cooled and poured into ice cold water than acidified and extracted with ethyl acetate, organic layer was washed with water and dried over anhydrous sodium sulfate, concentrate furnished 3-(3,4-dihydro-naphth-2- yl)-acrylic acid.
  • base such as piperidine using pyridine as solvent
  • the 3-(3,4-dihydronaphth-2-yl) ⁇ 2- alkyl-propenal was condensed with an ylide (Wittig reagent, prepared from triphenylphosphine and methyl/ethyl bromoacetate or methyl/ethyl chloroacetate) in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at temperature 5-80 0 C for 6-60 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like or benzene/toluene or dimethylformamide to yield corresponding 5-(3,4-dihydronaphth ⁇ 2-yl)-4-alkyl-2£,4is-pentadienoic acid ethyl ester which was saponified with a strong alkali solution using sodium hydroxide or potassium hydroxide followed by acidification to furnish 5-(3,4-dihydronaphth
  • the acid of formula Ib is converted to its tetrohydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and 1-5 5 atmospheric pressure to obtain corresponding tetrahydro derivative which was converted to corresponding amides of formula lba by the method described above for the preparation of compound of formula Ib.
  • polar solvent such as methanol or ethanol
  • lba may also be prepared by hydrogenation of compound of formula Ib as such by the method as described above.
  • the alpha-tetraione was first allowed to react with Grignard reagent (prepared from aryl halides & magnesium or unsubstituted or substituted- benzyl halide) with constant stirring at ambient temperature in an anhydrous ether solvent such as diethyl ether, tetrahydrofuran and the like to produce corresponding phenyl (unsubstituted or substituted) dihydronaphth- 2-yl-carbinol or benzyl (unsubstituted or substituted) dihydronaphthyl carbinol which was
  • the acid of formula Ib is converted to its i0 hexahydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and 1-5 atmospheric pressure to obtain corresponding tetrohydro derivative which was converted to corresponding amides of formula lba by the method described above for the preparation of compound of formula Ib.
  • polar solvent such as methanol or ethanol
  • lba may also be prepared by hydrogenation of compound of formula Ib as such by the method as described above.
  • R halogen, hydrogen, alkyl (Cl to ClO), substituted/unsubstituted aryl or benzyl.
  • RlR2R3R4 hydrogen, nitro, halo, hydroxy, alkyloxy, alkenyloxy, aryloxy, substitutedpyranyl radical unsubstituted pyranyl radical or Rl , R4 represnet hydrogen and R2,
  • R2+R3 1 ,3-dioxal, 1,4-dioxal
  • R5+R6 heterocyc!ic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-melhylplperazlnyl, oxazolyl, N-methylpipera ⁇ inyl, pyrrolyl, imidazolyl, oxazolyl, thiooxazole, an amino acid such as aianlnyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and Ih ⁇ like and optionally converting them to (heir salts by method known In the art of synthesis,
  • R7 normal or branched chain C1 to C10 alkyl groups or phenyl or benzyl radical branched chain) (substituted or unsubstituted)
  • Chemo resistance is a major problem in drug therapy.
  • the mechanisms underlying the clinical phenomena of de novo and acquired drug resistance may arise from alterations at any step in the cell-killing pathway. These include drug transport, drug metabolism, drug targets, cellular repair mechanisms and the ability of cells to recognize a harmful toxin or pathogen.
  • a common mechanism of reduced cellular drug accumulation is the increased expression Multi ⁇ Drug Resistance pumps (MDRs).
  • MDRs Multi ⁇ Drug Resistance pumps
  • the present invention also proves that the compounds described here inhibit the bacterial efflux, as a result of which there is more of accumulation and decreased efflux of ethidium bromide is used as substrate to gauge the efflux of the drugs by the extent of fluorescence observed.
  • the products of the present invention are novel mechanism based pharmaceutical entities acting through synergism and/or additive effect so that drugs contained in the tormulation are more bio-efficaceous as a result of one or more of the mechanism as revealed above and thereby increasing the sensitivity of the target cell to an anti-infeetive.
  • the Mrug' in the present invention refers to a chemical entity capable of affecting organism's patho-physiology and used for the treatment or prevention of disease.
  • Drugs include a number of classes of compounds, but not limited to aminoglycoside, penicillins, cephalosporins and other ⁇ -lactam agents, macrolides, glycopeptides, fluoroquinolones, tetracyclines, first and second line anti-TB drugs, anti-leprosy, antiviral, polyene, triazole, and imidazoles and combinations like pyrimidines, sulphamethoxazole.
  • Drugs may be a prodrug, activated or metabolised form, consisting of charged, uncharged, hydrophilic, hydrophobic or zwitter-ion species which make their entry by simple diffusion, carrier mediated transport dependent and not dependent on energy requirements, through ion and/ or voltage gated channels.
  • the 'potentiator' refers to aromatic amides of the formulae Ia, laa, Ib, lba selected from the following set of compounds prepared by the method described in the examples.
  • 3-(l-benzyl-3,4-dihydro-napth-2-yl)-acrylic acid pyrrolidide 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid morpholide 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisopropylamide 3 -( 1 -benzyl-3 ,4-dihydronapth-2-yl)-acrylic acid ⁇ -hydroxy piperidine 5 3 -(6,7-dimethoxy-3 ,4-dihydronaphth-2-yl)- ⁇ ropenoic acid piperidide
  • a therapeutically effective amount means those amounts of the efflux pump inhibitor and anti-microbial agent which, when used in combination produce the desired therapeutic effect as judged by the model animal infection studies.
  • the efflux pump inhibitor and anti-microbial agent are combined in predetermined proportions and thus a therapeutically effective amount would be an amount of the combination and the amount of individual components i.e.
  • efflux pump inhibitor and the anti-microbial agent as well as the combination of two can be determined by one of the skill in the art and the amount of the combination will vary depending upon several factors such as the particular microbial strain /isolate used in the study and the particular efflux pump inhibitor and the anti-microbial agent used besides depending upon the weight, sex, age etc of the animal/patient on whom the combination of the efflux pump inhibitor and the anti-microbial agent is tested.
  • a pharmaceutical carrier is generally an inert bulk agent added to make the ingredients achieve superior ad mixing and can be solid or liquid. The inert parts of standard pharmaceutical compositions used in this process are also part of the present invention.
  • the checker board method is a checker board method
  • checkerboard refers to the pattern (of tubes or microtiter plate wells) formed by multiple dilutions of two drugs being tested (Eliopoulos GM, Moelle ' ring RC. Antimicrobial Combinations, in: Antibiotics in Laboratory Medicine: USA: Williams & Will ⁇ ns).
  • the checkerboard consisted of columns in which each tube (or well) contains the same amount of the standard drug (antibacterial/antifungal/anti-TB/antiviral) being diluted along the x-axis and rows in which each tube (or well) contains the same amount of the potentiator being diluted on the y-axis.
  • each square in the checkerboard (which represents one tube/ well or plate) contained a unique. This chequerboard technique can be performed with liquid or semisolid (agar) media.
  • the agar (Mueller Hinton agar, Middlebrook 7H10 agar) was autoclaved and allowed to cool to 55 0 C to 50 0 C.
  • the combination of the standard drug and the potentiator was added to the agar.
  • Serial two fold dilutions of each of standard drug and the potentiator were prepared in appropriate solvents.
  • the volume of solvent (containing standard drug or potentiator) added to agar was kept small (i.e. 5% of the total volume).
  • the bacteria to be tested were applied to the surface of agar with a replicating device designed to deliver a standard inoculum (approx 10 4 cfu
  • the plates were incubated at 37 0 C for 24 hrs (3 weeks in case of Mycobacterium tuberculosis).
  • R 7 methyl
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • Example 69d Decrease in the MICs of Amikacin against Staphylococcus aureus, MRSA and . other Gram positive bacteria when used in combination with compound of formula Ia where Minimum Inhibitory Concentration (MIC) of Amikacin alone and in combination with the above mentioned potentiator was performed against Staphylococcus aureus species, using method described in the study design. Sixteen to four fold reductions in MIC of Amikacin was observed in combination with the potentiator (Table Id).
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • MIC Minimum Inhibitory Concentration
  • Staphylococcus aureus ATCC 29213 (10 7 CFU/mouse).
  • the infected mice were divided in groups and each group consisted of 6 mice.
  • the treatment consisted of one dose immediately after the infection followed by the next dose after a gap of 6 hrs.
  • the result was recorded as number of survivals each day.
  • the mice were observed for seven days and ED50 was determined after seven days of observation.
  • the ED 50 for ciprofloxacin was 9.2
  • bacterial suspensions were prepared as described above and were exposed to ethidium bromide (2 ⁇ g/ml) in the presence of compound of formula 1 (25 ⁇ g/ml) for 30 min at 37 0 C. The cells were then pelleted by centrifugation and were re-suspended in fresh uptake buffer. The loss of ethidium bromide from the cells was measured as a decrease in fluorescence ( Figure IB).
  • the present invention discloses the preparation of a novel class of compounds Le 3-( unsubstit ⁇ ted-or-substituted-3,4-dihydronaph-2yl-)-acrylic acid amides,5— (unsubstituted-or substituted-3,4-dihydronaph-2yl-)4-substitued-2E,4E-pentadionic acid amides and the saturated derivatives of these compounds ,the whole class of compounds not reported hitherto in the literature and their use as potentiators of antibiotics (i.e as efflux pump inhibitors of bacteria) when used in combination with antibiotic and tested against bacteria.
  • antibiotics i.e as efflux pump inhibitors of bacteria
  • the synthesised molecules have shown high potency in terms of lowering of MIC of the drug ciprofloxacin (thirtytwo fold) compared to the MIC levels of drug when used alone. These classes of compounds have shown potentiation of the in-vivo efficacy of the drugs when tested in in-vivo animal experimental models.

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Abstract

This invention relates to compounds, which are in general potentiators of antiinfeective agents and inhibitors of bacterial efflux pumps in particular. These novel compounds i.e. substituted dihydronaphtalene derivatives and tetralin derivatives and/or their salts are useful against antibiotic-resistant microbial pathogens, for treating microbial infections by reducing the extrusion of a co-administered antimicrobial agent. This invention also provides processes for the preparation of such efflux pump inhibitors and their compositions with anti infective agents and methods for treatment of microbial infections.

Description

"NOVEL EFFLUX PUMP INHIBITORS"
Field of the invention
The present invention relates to the field of chemotherapeutics, particularly use of novel synthetic aromatic amides for potentiating the bio-efficacy of specific drugs. The invention particularly relates to the preparation and use of synthetic analogues of aromatic amides, useful in potentiating bio-efficacy of anti-infective drugs. The invention more particularly relates to aromatic substituted acrylic acid amide of general formula 1 including its analogues or and salts thereof
Figure imgf000002_0001
Formula-1
Wherein n~ 1 or 2, dotted lines indicates the single or double bond, wherein n=l R7 - H wherein n=2 R7 = normal or branched chain Cl to ClO alkyl group, phenyl, benzyl radical at 4-position and R7=H at C-2 position, wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO) or aryl or benzyl (unsubstituted and substituted); R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy* aryloxy, substituted pyranyl radical, unsubstituted pyranyl radical or R1, R4 represents hydrogen group and R21R3 represent hydrogen ,alkoxyl(Cl-C10), alkenoxyl groups (Cl-ClO), aryloxyl, substituted aryloxyl groups or Ri, R4 represents hydrogen group and R2+R3 together represent 1,3-dioxol , 1,4-dioxol; R3- represents hydrogen atom or normal or branched chain C1 to C10 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain Ci to Cm alkyl group or phenyl or benzyl radical; where NR5R6 together (R5+R6) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N- methylpiperazinyl, oxazolyl, N-methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like and optionally converting them to their salts by method known in the art of synthesis. The novel amide when administered in combination with an anti-infective drag potentiates the bioactivity of the drug thereby requiring lower doses and/or decreased frequency of dosing while maintaining the therapeutic efficacy of standard doses of such drugs. The synthetic molecules of this invention are all novel, as they have not been reported 5 earlier in the literature. These novel molecules possess specific properties of potentiating the bio-efficacy of specific drugs particularly they are useful in combination with anti-infective drugs which besides reducing their effective dosages are also associated with lesser side effects. The molecules along with the pharmaceutical products/combinations possessing the properties disclosed in the present invention are novel and not known in literature or prior LO art.
Background of the invention
For preventive or therapeutic treatment of infectious diseases caused by micro-organisms, various antibacterial agents have so far been developed, and drags such as β -lactam
[5 antibiotics (penicillins, cephems, monobactams, carbapenems, and penems), aminoglycosides, quinolones, macrolides, tetracyclines, rifamycins, chloramphenicols, and phosphomycins have been practically used. However, with the increase of clinically used amount of antibacterial agents, remarkable numbers of resistant bacterial strains to these antibacterial agents have emerged, which becomes a serious problem in the treatment of
>0 infectious diseases.
There are three proven targets for the main antibacterial drags: (1) bacterial cell-wall biosynthesis; (2) bacterial protein synthesis; and (3) bacterial DNA replication and repair. There are number of mechanisms through which the micro-organisms resistance against these therapeutic agents. These mechanisms include target overproduction or modification,
J5 permeability barriers (reduced uptake or active efflux), enzymatic inactivation, and sequestration.
One of the most frequently employed resistance strategies in both pro-karyotes and euk, yotes is the transmembrane efflux pump proteins, which are involved in the extrusion of toxic substrates (including virtually all classes of clinically relevant antibiotics) from
SO within cells into the external environment. These proteins are found in both gram-positive and gram-negative bacteria as well as in eukaryotic organisms (F. Van Bambeke, E. Balzi,
? P.M. Tulkens, Biochem. Pharmacol. 60 (2000) 457-470 Pumps may be specific for one substrate or may transport a range of structurally dissimilar compounds (including antibiotics of multiple classes); such pumps can be associated with multiple drug resistance (MDR). In the prokaryotic kingdom, there are five major families of efflux transporter (O. 5 lomόvskaya, M.. S. Warren. A; Lee, J. Galazzo, R1 fronko, M. Lee, J, Blais, D. Cho, S. Chamberland, T.ranau, R. Leger, S. Hecker, W. Watldns, K. oshino, H. ishida, V. J. Lee., Antimicrob. Agents and Chemother. 2001, 45: 105-116. MF (major facilitator), MATE (multidrug and toxic efflux), RND (resistance-nodulation-division), SMR (small multidrug resistance) and ABC (ATP binding cassette). All these systems utilize the proton motive
10 force as an energy source (LT. Paulsen, Microbiological Reviews, 1996, 60: 575-608 apart from the ABC family, which utilizes ATP hydrolysis to drive the export of substrates. In gram-positive bacteria, MDR is mainly conferred by MFS efflux systems, the most studied pumps being Nor A of S. aureus (A. A. Neyfakh, C.M.Borsch, G.W.Kaatz Antimicrob. Agents and Chemotherapy, 1993, 37: 128-129) and its homologues in B.
15 subtilis, Bmr and Bit (A. A. Neyfakh, Antimicrob. Agents and Chemotherapy, 1992, 484- 485). For gram-negative bacteria, RND efflux systems are major contributors to resistance: AcrAB-TolC and MexABOprM are involved in intrinsic resistance of E. coli (M. C. Sulavik. C. Houseweart, C.cramer, N. Jiwai, N. Murgolo, J. Greene, B. Didomenico, K. J. Shaw, G. H. Miller, R. Hare, G. Shinier Antimicrob. Agents and Chemotherapy, 2001, 45:
10 1126-1136) and P. aeruginosa (X. Li, N.Nikaido,L.Poole Antimicrob. Agents and Chemotherapy, 1995, 35: 1948-), respectively. One strategy to target these microbial efflux pumps is to find inhibitors of these efflux pumps and, in particular of bacterial and fungal efflux pumps. Recently, we have demonstrated, the preparation and the application of newer class of
.5 synthetic molecules as potentiators of bioefficacy of antiinfectives (S. Koul, J.L. Koul, S. C. Taneja, P. Gupta, LA. Khan, Z.M. Mirza, A. Kumar, R.K. Johri, M. Pandita, A. Khosa', A.K. Tikoo, S.C. Sharma, V. Verma, and G.N. Qazi. U.S. PGPUB-0004645), where in the inhibitors disclosed are 4-alkyl-5-substituted phenyl 2£,4.£-pentadienoicacid amides and its tetra hydro analogies which is unlike the efflux pump inhibitor compounds of the present
$0 invention. It may be said here that in the above mentioned U.S. patent 20070004645 the potentiation of the drugs like ciprofloxacin and mupirocin in combination with most potent compounds like 5-(3,4-methylenedioxyphenyl)-4-ethyl-2E,4E-pentadienoic acid piperidide & 5-(3,4-dimethoxyphenyl)-4-ethyl-2E,4E-pentadienoic acid piperidide) showed up to eight fold reduction in the MIC of the said drugs. Whereas, the present invention discloses up to sixteen fold reduction in the MIC of the above said drugs (as shown in the examples 69a to 5 75 and tables 1 to 5 of the present invention).
Objects of the invention
The main object of the present invention is to provide the novel 3-(l-substituted-3,4- dihydro-naphth-2-yl)-acrylic acid amide, its tetra hydro derivatives of general formula Ia, 10 laa and 5-(l-substituted-3,4-dihydronaphth-2-yl)-4-alkyl-2E,4E-pentadienoic acid amides its tetrahydro derivatives of structure formulae Ib, lba its analogues and /or their salts thereof.
Another object of the invention is to provide the novel aromatic substituted pentadienoic acid amide, which may be useful as potentiators of the bioefficacy of the drugs. 15 Yet another object of the invention is to provide the process for the preparation of the novel aromatic amides.
Still another objective of the invention is to provide the compounds of formula 1, which are not toxic.
Further object of the invention is to provide the pharmaceutical composition using effective ZO amount of one or more compound of formula Ia including its geometrical isomers, its analogues or and salts thereof as stated above along with the anti-infective drug and optionally along with a carrier or diluent or pharmaceutically acceptable exciepient.
Another object of the invention is to provide the pharmaceutical composition, which is useful for the treatment of the infections caused by bacteria. .5 Further object of the invention is to provide the pharmaceutical composition which is used in the reduction in the dose requirement of anti-infectives.
Summary of the invention
The present invention provides a new class of bacterial efflux pump inhibitors(EPIs) (not »0 reported in the literature) for therapeutic use when co-administered with antiinfectives (antibiotics).The EPIs as per the present invention belongs to the general formula 1
Figure imgf000006_0001
Formula-1
Figure imgf000006_0002
1a 1aa
Figure imgf000006_0003
1 b 1ba wherein the compounds falling under the general formula laa,lab,lba and laa belong to 3-
(l-substituted-3,4-dihydro-naphth-2-yl)-acrylic acid amide, its tetra hydro derivatives (general formula Ia, . laa) and 5-(l-substituted-3,4-dihydronaphth-2-yl)-4-alkyl-2E,4E- pentadienoic acid amides its tetrahydro derivatives (general formulae Ib, lba) its analogues and /or their salts thereof. Strains of Staphylococcus aureus 1199 and its mutant Nor A over expressing Staphylococcus aureus 1199B of the genus Gram +ve bacteria known for its resistance to a host of antibiotics (including Ciprofloxacin) when treated with a formulation comprising of antibiotic ciprofloxacin and an EPI (selected from the compounds of the formula 1) in a given definite ratio by potentiation of the drug activity which is because of the inhibition of the efflux pump present in the bacteria. This is evidenced by the experiments shown in the present invention using ethidium bromide (EtBr) which is the substrate of efflux' pump and is thrown out as evidenced by the fast loss of fluorescence of EtBr whereas in presence EPI there is retention of the fluorescence of EtBr and no fluorescence depletion is observed.
The present invention describes a new class of bacterial efflux pump inhibitors (EPIs) for therapeutic use when given in combination with antibiotics. The EPIs as per the present invention are selected from the general formula 1 which contains compounds that belong to 3-(l-substituted-3,4-dihydro-naphth-2-yl)-acrylic acid amide, its tetra hydro derivatives (general formula Ia, laa) and 5-(l-substituted-3,4-dihydronaphth-2-yl)-4-alkyl-2E,4E- pentadienoic acid amides its tetrahydro derivatives (general formulae Ib, lba) its analogues and /or their salts thereof. A formulation comprising of an antibiotic such as ciprofloxacin is co-administered with an EPI selected from the above mentioned class of compounds and tested against drug resistance bacterial strains such as Staphyllococcus aureus 1199 and its mutant Nor A over expressing Staphyllococcus aureus 1199B (Gram +ve bacteria known for its resistance to aJiost of antibiotics including Ciprofloxacin,mupirocin ). The drug potency is restored due to the inhibition of the bacterial efflux pump as evidenced by the experiments using ethidium bromide (EtBr) where the EPI inhibits the fast loss of fluorescence of EtBr which in absence of EPI is depleted fast.
In the present invention, preparation and application of a new class of bacterial efflux pump inhibitors (EPIs) for therapeutic use is described when given in combination with antibiotics. The EPIs as per the present invention are selected from the general formula 1 which contains compounds that belong to 3-(l-substituted-3,4-dihydro-naphth-2-yl)-acrylic acid amide, its tetra hydro derivatives (general formula Ia, laa) and 5-(l-substituted-3,4- dihydronaphth-2-yl)-4-alkyl-2£,4ii-pentadienoic acid amides its tetrahydro derivatives (general formulae Ib, lba) its analogues and /or their salts thereof. A formulation comprising of an antibiotic such as ciprofloxacin is co-administered with an EPI selected from the above mentioned class of compounds and tested against drug resistance bacterial strains such as Staphyllococcus aureus 1199 and its mutant Nor A overexpressing Staphyllococcus aureus 1199B (Gram +ve bacteria known for its resistance to a host of antibiotics including Ciprofloxacin,mupirocin ). The drug potency is restored due to the inhibition of the bacterial efflux pump as evidenced by the experiments using ethidium bromide (EtBr) where the EPI inhibits the fast loss of fluorescence of EtBr which in absence of EPI is depleted fast as shown in Figure- IA and Figure- IB Brief description of the drawings
Figure-IA Accumulation of ethidium bromide in wild type and mutant strain, CIPr-I and effect of compound of formula Ia corresponds to compound 3-(l-chloro-3,4-dihydronapth- 2-yl)-acrylic acid N5N-diisopropylamide at 25 μg/ml.
In the figure-IA, the compound of formula I was added after 30 min (as indicated by the arrow in the figure) of the start of the experiment. This addition resulted in the increased accumulation of ethidium bromide (due to the inhibition of the efflux pump) in the wild and mutant strains as indicated by the increase in the fluorescence intensity.
Figure-IB Efflux of ethidium bromide in wild type and mutant strain. ClPr-l and effect of compound of formula Ia corresponds to compound 3-(l-chloro-3,4-dihydronapth-2-yl)- acrylic acid N,N-diisopropylamide at 25 μg/ml.
Figure-IB depicts the inhibition of efflux of ethidium bromide from the wild and mutant strain of S. aureus when pretreated with the compound of formula Ia, resulting in no loss of fluorescence compared to controls where the compound of formula I is not present, which leads to rapid decrease in the fluorescence.
Description of the invention
Accordingly the present invention provides a substituted aromatic amides of general formula Ia, Ib, laa, lba and salts thereof as potentiators of bioefficacy of anti-infective drugs when used in combination with anti-infective drugs in the form of a pharmaceutical composition.
Figure imgf000008_0001
Ia, Ib, laa and lba
Wherein n= 1 or 2, dotted lines indicates the single or double bond, wherein n=l R7 = H wherein n=2 R7 = normal or branched chain Cl to ClO alkyl group, phenyl, benzyl radical at 4-position and R7=H at C-2 position, wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO) or aryl or benzyl (imsubstituted and substituted); R1,R25R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, substituted pyranyl radical, unsubstituted pyranyl radical or Rl, R4 represents hydrogen group and R2, R3 (R2=R3) represent hydrogen , alkoxyl (Cl-ClO) , alkenoxyl groups (Cl- ClO) , aroyloxyl or substituted aroyloxyl groups or Rl, R4 represents hydrogen group and R2 + R3 together represent 1,3-dioxol or 1,4-dioxol;
R5 represents hydrogen atom or normal or branched chain Ci to Cio alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain C1 to C10 alkyl group or phenyl or benzyl radical; where NR5R6 together (R5H-R6) represent heterocyclic amine radical " such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N- methylpiperazinyl, oxazolyl, N-methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like and optionally converting them to their salts by method icnown in the art of synthesis. The structural formula of compound Ia, Ib, laa and lba, represented as follows :
Figure imgf000009_0001
Ia
Figure imgf000009_0002
laa
Formula-la and laa, wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO), aryl, benzyl (unsubstituted and substituted);
R1,R2,R3,R4 is hydrogen, nitro, halogen , hydroxy, alkyloxy, alkenyloxy, aryloxy, substituted pyranyl radical ,unsubstituted pyranyl radical or Rl , R4 represents hydrogen group and R2, R3( R2=R3) represent hydrogen , alkoxyl (Cl-ClO) or alkenoxyl groups (Cl- ClO) also aryloxyl or substituted aryloxyl groups or Rl, R4 represents hydrogen and R2+R3 together represent 1 ,3-dioxol or 1 ,4-dioxol,;
R5 represents hydrogen atom or normal or branched chain Ci to C10 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain C1 to Cjo alkyl group or phenyl or benzyl radical; where NR5R6 together (R3-+R6) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N- methylpiperazinyl, oxazolyl, N-methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like.
Figure imgf000010_0001
Ib
Figure imgf000010_0002
lba
Formula-lb and lba, wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO), aryl , benzyl (unsubstituted and substituted);
R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, substituted pyranyl radical ,unsubstituted pyranyl radical or Rl, R4 represents hydrogen group and R2, R3(R2=R3) represent hydrogen, alkoxyl(Cl to ClO), alkenoxyl groups (Cl- ClO)5 aryloxyl, substituted aryloxyl groups or Rl, R4 represents hydrogen group and R2+R3 together, represents 1 ,3-dioxol or 1 ,4-dioxol,; R5 represents hydrogen atom or normal or branched chain Ci to C]0 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain Ci to Cio alkyl group or phenyl or benzyl radical; where NR5R6 together (R5+R6) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl, N-methylpiperazinyl., pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like, wherein R7 represents normal or branched chain Cl to ClO alkyl group.
In an embodiment of the invention wherein the salt is pharmaceutically acceptable selected from hydrochloride, acetate, succinate, maleate.
Accordingly the present invention provides a process for the preparation of substituted aromatic acid amides of general formula Ia, laa, Ib and lba as stated above, the method of preparation comprising;
(i) subjecting aldehyde compound of formula 2,
Figure imgf000011_0001
Formula 2 Formula 3 wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Ci to Ci0) or aryl or benzyl (unsubstituted and substituted); Ri,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, substituted pyranyl radical, unsubstituted pyi'anyl radical or Ri1 R4 represents hydrogen group and R25R3 (R2=R3) represent hydrogen, alkoxyl ' (CI-CI0), alkenoxyl groups (Ci-Cio), aryloxyl, substituted aryloxyl groups or Ri1 R4 represents hydrogen group and R2+R3 together represent 1,3-dioxol or 1,4-dioxol to Wittig reaction (Wittig reagent prepared from triphenylphosphine and ethyl bromoacetate/ ethylchloroacetate in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at temperature 5-80 0C for 1 to 24 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like benzene/toluene or dimethylformamide) and subsequent saponification of the Wittig reaction product to get compound of the formula 3, (ii) alternately allowing aldehyde of the formula 2 to react with malonic acid in presence of a base like piperdine and pyridine, contents stirred at temperature 25-40 0C for 24-30 hours followed by subsequent decarboxylation carried out by heating at 80-90 0C, acidification of the reaction mixture by dilute HCl, or H2SO4 or the like, extraction of the contents, concentration of the organic layer, to get the compound of formula 3, (iii) treating the compound of formula 3 with thionylchloride and subsequent reaction of the acid chloride thus prepared with an appropriate amine to get the compound of formula Ia by conventional method wherein RiR2R3R4 is hydrogen, nitro, halogen , hydroxy, alkyloxy, alkenyloxy, substituted pyranyl radical,un-substituted pyranyl radical or R1, R4 -represents hydrogen group and R2 , R3 (R2=R3) represent hydrogen , alkoxyl (Ci-C10) or alkenoxyl groups (Ci-Cio) , aryloxyl , substituted aryloxyl groups or Ri; R4 represents hydrogen group and R2 + R3 together represent 1,3-dioxol or 1,4-dioxol, ;R5 represents hydrogen atom or normal or branched chain C1 to Ci0 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain C1 to Ci0 alkyl group or phenyl or benzyl radical; where NR5R6 together (R5+R6) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl, N- methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and optionally converting them to their salts by known method by reacting with an acidic compounds such as hydrochloric acid, acetic acid, tartaric acid,
Figure imgf000012_0001
Formula 1 a
(iv) converting the compound of formula Ia to its tetrahydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature >0 and at 1-3 atmospheric pressure to obtain corresponding tetrahydro derivative of general formula laa,
Figure imgf000012_0002
Formula laa
(v) alternately, hydrogenation of compounds of the formula 3 with Pd/charcoal/ solvents5 such as methanol, ethanol, ethylacetate and at 1-3 atmospheric pressure and treatment of the tetrahydro derivative with thionyl chloride and followed by treatment of the acid chloride with appropriate amine afforded compound of the formula Iaa wherein R,R1,R2,R3,R4,R5,R6 are radicals/groups as described above for the compound of the formula Ia,
(vi) For the preparation of compound of the formula Ib the method of preparation comprises of reacting compound of the formula 2 with Grignard reagent prepared from appropriate alkyl halide (represented by R7X, chosen from Cl-ClO normal or branched alkylhalide, substituted or unsubstituted benzyl or aryl halides) and Mg metal with constant stirring at ambient temperature in an anhydrous ether/THF solvent to produce compound of the formula 4 wherein R11R2 R31R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, substituted pyranyl radical, unsubstituted pyranyl radical or R1; R4 represents hydrogen group and R2,R3 (R2=R3) represent hydrogen atoms or alkoxyl(Cl to ClO) or alkenoxyl groups (C]-Cio), aroyloxyl, substituted aroyloxyl groups or Ri, R4 represents hydrogen group and R2+R3 together represent 1,3-dioxol or 1,4-dioxol and R7 is Cl-ClO normal or branched alkylhalide, substituted or unsubstituted benzyl or aryl halides,
Figure imgf000013_0001
Formula Ib
(vii) The compounds of formula -4 treated with Vilsmeier reagent comprising of dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tribromide mixture at 0-50 0C preferably at 0-10 0C for 5-50 hrs. Preferably for 5-12 hrs, followed by dilution of contents with ice cold water, bringing the pH to 7 by neutralisation with dilute alkali solution to produce compound of formula 5 wherein R1,R2,R3,R4 and R7 are radicals or substituents as described in for compounds of the formula 4,
Figure imgf000013_0002
Figure imgf000013_0003
Formula 5 (viii) condensation of the compounds of the formula 5 with an ylide (Wittig reagent, prepared from triphenylphosphine and ethyl bromoacetate/ ethylchloroacetate) in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at a temperature ranging between 5-80 0C for a period ranging between 1 to 24 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like, or benzene/toluene or dimethylformamide followed by direct saponification of Wittig reaction product furnished compounds of the formula 6 wherein R11R2R31R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, allcenyϊoxy, substituted pyranyl radical, unsubstituted pyranyl radical or Ri1 R4 represents hydrogen group and R2,R3 represent hydrogen, alkoxyl (C1-Ci0), alkenoxyl groups (Ci-Cio), aryloxyl, substituted aryloxyl groups or R)1 R4 represents hydrogen group and R1+R2 together represent 1,3-dioxol or 1,4-dioxol; and R7 is Cl-ClO normal or branched alkylhalide, substituted or unsubstituted benzyl or aryl radical,
Figure imgf000014_0001
Formula 6 (ix) treating the compound of the general formula 6 with thionyl chloride in presence of a base such as pyridine followed by treatment of the acid chloride with an appropriate base afforded compounds of the formula Ib wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO) or aryl or benzyl (unsubstituted and substituted); R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, sxibstituted pyranyl radical, unsubstituted pyranyl radical or Rl, R4 represents hydrogen group and R2, R3 represent hydrogen, alkoxyl (Cl-ClO), alkenoxyl groups (Cl-ClO)1, aryloxyl, substituted aryloxyl groups or Rl, R4 represents hydrogen group and R2+R3 together represents 1,3-dioxol or 1,4-dioxol; R5 represents hydrogen atom or normal or branched chain C1 to C10 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain Ci to Ci0 alkyl group or phenyl or benzyl radical; where NR5R6 together (R5+R5) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl, N-methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like, wherein R7 represents normal or branched chain Cl to ClO alkyl group,
Figure imgf000015_0001
Formula Ib
(x) converting the compound of formula Ib to its tetra hydro derivative by hydro genation in presence of Pd/charcoal in polar solvent such as methanol or ethanol or ethyl acetate at ambient temperature and at a pressure ranging between 1-3 atmosphere to obtain compound of formula lba.
Figure imgf000015_0002
Formula lba
(xi) alternately, hydrogenation of compounds of the formula 6 with Pd/charcoal/ solvents such as methanol, ethanol, ethyl acetate and at a pressure ranging between 1-3 atmosphere and subsequent treatment of the tetrahydro derivative with thionyl chloride followed by treatment of the acid chloride thus obtained with appropriate amine afforded compound of the formula lba wherein R,R1,R2,R3,R4,R5,R6,R7 are radicals/groups as described above for the compound of the formula Ib.
In an embodiment of the invention wherein the use of compound of formula 1 that include compounds of the formula la,laa,lb and lba as potentiator of bioefiϊcacy of anti-infective drugs.
In another embodiment of the invention wherein the process further comprises converting the compound of formula Ia,laa4b and lba into a salt.
Accordingly, the present invention provides a pharmaceutical composition comprising an effective amount of one or more compound of formula la,laa,lb and lba its analogues or /and salts thereof as stated above along with the anti-infective drug and optionally along with a carrier or diluent or pharmaceutically acceptable exciepient.
In an embodiment of the invention the amount of compound of formula 1 may vary from 0.1 to 50% by weight of the composition with respect to the drug. In another embodiment of the invention wherein the composition is useful as an anti - microbial agent.
In yet another embodiment of the invention wherein the MIC value of composition is reduced more than 32 times of the anti- infective drugs when used alone. In an another embodiment of the invention wherein the ED50 of anti-infective drug reduced to about 1/4 when used in combination with the compound of formula Ia, laa, Ib and lba.
In still another embodiment of the invention wherein the dose of anti-infective drugs is reduced about 1A when used in combination with compounds of formula la,laa,lb and lba.
In further embodiment of the invention wherein the anti-infectives are selected from groups comprising of penicillins (including semi-synthetic), cephalosporins, aminoglycosides, glycopeptides, fluroquinolones, macrolides, tetracyclines other antibiotic groups such as mupirocin and framycetin, first and second line anti-TB drugs, anti-leprosy ' drugs, oxazolidinones.
In an embodiment of the invention wherein the composition is effective against micro- organisms selected from gram positive bacteria and gram negative bacteria, that include
Staphylococcus species, Bacillus species. Pseiidomonas species, E coli and Salmonella species as well as against micro-organisms such as Mycobacterium species.
In another embodiment of the invention wherein composition is effective against growth of micro-organisms that include anti-infective resistant strains such as MRSA. In a further embodiment of the invention wherein composition is effective when tested for curing of mice, guinea pig or rabbit models infected with micro-organisms from the gram positive group of bacteria and gram negative group of bacteria such as Staphylococcus species,
Bacillus species., Pseiidomonas species, E coli, Salmonella species besides being effective against microorganisms such as Mycobacterium species. In an embodiment of the invention wherein the said composition along with excipient or pharmaceutical vehicle such as lactose, corn starch, polyethylene glycols and the like may be administered through oral route.
In an embodiment of the invention wherein the said composition along with excipient or pharmaceutical vehicle such as castor oil, olive oil and the like may be administered through systemic route.
In an embodiment of the invention wherein the said composition along with excipient or pharmaceutical vehicle such as polyethylene glycols, bees wax, paraffin wax, emulsifying agents and the like may be applied topically.
A method for inhibiting a bacterial cell that employs an efflux pump resistance mechanism, comprising contacting the cell with a pharmaceutical composition comprising of an antibacterial agent and a compound of formula la,laa,lb,lba and optionally an excipient or pharmaceutical vehicle.
In an embodiment of the present invention wherein the bacterial cell is selected from gram positive bacteria such as Staphylococcus species and Bacillus species.
In one more embodiment of the invention wherein the bacterial cell is selected from gram negative bacteria such as Pseudomonas species, E coli and Salmonella species.
In a further embodiment of the invention wherein the bacterial cell is selected from mycobacterial species.
A method of treating infections comprising administrating to a subject in need of such treatment a therapeutically effective amount of compound of formula Ia, laa, Ib, lba, its analogues or/ and salts thereof as claimed in claim 1 along with the anti-infective drug and optionally along with a carrier or diluent or pharmaceutically acceptable exciepient.
In one of the embodiment of the invention wherein the infection caused by a bacteria that employs an efflux pump resistance as one of the ways of resistance, comprising administering to a patient in need thereof a therapeutically effective amount of an antibacterial agent and a compound of formula Ia, laa, Ib and lba.
The present invention deals with such combinations comprising 3-(l-substituted-3,4- dihydro-naphth-2-yl)-acrylic acid amide of structural formula Ia, including its related tetralin derivatives of structural formulae laa and 5-(l-substituted-3,4-dihydronaphth-2-yl)- 4-alkyl-2E,4E-pentadienoic acid amide of formula Ib including its related tetralin derivatives of structure formulae lba where R1 to R7 are as described above and as a representative example of formula 1 where R=Cl, Ri=R2=R3=R4=H and R5+R6=isobutylamine (example-5) and N,N-diisopropylamine (example-8), displayed the properties of potentiation/synergism in in vitro screening when combined with various anti- infective agents using bacteria, viruses and yeast. These combinations were also efficacious when tested in in vivo using mice and guinea pig models infected with microorganisms (example 69d-f and 73 a) and displayed the properties of potentiation/synergism when combined with various anti-infective agents in vitro using bacteria, viruses and yeast. The present invention is aimed to circumvent such problems and use of the products of the present invention offer a low dose regimen that produces enhanced therapeutic action comparable to that of standard dose of a drug alone. The compounds of the present invention are neither reported in the chemical literature nor have been used for the purpose of potentiating the bio-efficacy of the drugs particularly anti- infective drugs such as described in the present invention. The synthesis of the novel compounds have been accomplished through the combination of various chemical steps known in the art of synthesis though not for the purpose of synthesising the novel compounds of formula Ia, laa, Ib and lba as described above.
The aromatic amide of formula 1 i.e. 3-(l-substituted-3,4-dihydronaphth-2-yl)-acrylic acid amide, including its related tetralin derivatives of structural formulae Ia and 5-(l- substituted-3,4-dihydronaphth-2-yl)-4-alkyl-2E,4E-pentadienoic acid amide of formula Ib including its tetralin derivatives of structure formulae lba where R and R1 to R7 are described as above, were synthesised from the corresponding Tetralone and eugenol and their derivatives by the known state of art as described in literature. Method-1
Alpha.-tetralone (1,2,3,4-tetrahydronaphtha-l-one) was first reacted with Vilsmeier reagent comprising of dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tribromide mixture at 0-10 0C for 5-12 hrs, contents were neutralised with dilute alkali solution to produce l-halo-2-formyl-3,4-dihydronaphthalene. The l-halo-2-formyl- 3,4-dihydronaphthalene was condensed with an ylide (Wittig reagent, prepared from triphenylphosphine and ethyl bromoacetate/ethylchloroacetate) in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at temperature 5 - 80 0C for 1 to 24 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like benzene/toluene or dimethylformamide to yield corresponding 3-(l-halo-3,4-dihydro-naphth-2-yl)-acrylic acid ethyl ester which was hydrolysed with a strong alkali solution using sodium hydroxide or potassium hydroxide followed by acidification to furnish 3-(l-halo-3,4-dihydronaphth-2-yl) acrylic acid. A solution of aryl alkenoic acid as prepared above in an inert organic solvent such as benzene, dichloromethane was treated with thionyl chloride and excess of solvent removed in vacuo. The acyl chloride intermediate thus obtained without purification was condensed with acyclic or cyclic or heterocyclic amine in inert organic solvent such as dichloromethane, benzene, diethyl ether and the like in the temperature range of 5-50 0C, and after the purification by crystallisation or column chromatography to produce amide of formula 1. For the preparation of compound of formula Ia, the acid of formula 1 was converted to its tetrahydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and at 1-3 atmospheric pressure to obtain corresponding tetrahydro derivative which was converted to corresponding amides of formula Ia by the method described as above for the preparation of compound of formula 1. Alternatively Ia may also be prepared by hydrogenation of compound of formula 1 as such by the method as described above. .
Method-2
Eugenol l-(4'-hydroxy-3'-methoxy-phenyl)-2-propene was first reacted with alkyl/alϊyl/aryl alkyl halide using K2CO3 and dry acetone mixture at reflux temperature for 24-50 hrs, the reaction mixture worked up by conventional method and the crude product purified by column chromatography over silica gel using hexane-ethyl acetate mixture as eluent to give the product as yellow viscous liquid. The eugenol alkyl ether was then reacted with dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tri bromide mixture (N.S. Narasimhan, T. Mukhotadhyay, Teterahedron Lett. (1979) 1341- 1342. & N.S. Narasimhan, T. Mukhotadhyay, S.S. Kusurkar, Indian J. Chem. 2OB (1981) 546-548) at 0-10 0C for 40-72 hrs, contents neutralised with dilute alkali solution to produce 6,7-dia'lkoxy-3,4-dihydronaphthalene-2-carbaldehyde. The 6,7-dialkoxy-3,4 dihydronaphthalene-2-carbaldehyde was then condensed with malonic acid in presence of base such as piperidine using pyridine as solvent and the reaction left at room temperature for 24-30 hrs after that heated at 80-900C5 contents were cooled and poured into ice cold water than acidified and extracted with ethyl acetate, organic layer was washed with water and dried over anhydrous sodium sulphate, concentrate furnished 3-(6,7-dialkoxy-3,4- dihydro-naphth-2-yl)-acrylic acid.
A solution of aryl alkenoic acid as prepared above in an inert organic solvent such as benzene, dichloromethane treated with thionyl chloride and excess of solvent removed. The acyl chloride intermediate thus obtained without purification was condensed with acyclic or cyclic or heterocyclic amine in inert organic solvent such as dichloromethane, benzene, diethyl ether and the like in the temperature range of 5-50 0C, and after the purification by crystallisation or column chromatography to produce amide of formula Ia.
Method-3 The tetralone was first reacted with dimethylformamide and phosphorus oxybromide or phosphorus tri bromide mixture at 0-10 0C for 5-24 hrs, contents neutralised with dilute alkali solution to produce l-halo-3,4-dihydronaphthalene-2-carbaldehyde . The l-halo-3,4- dihydronaphthalene-2-carbaldehyde was reacted with alkyl magnesium halide (alkyl halide C2-C1O) with constant stirring at ambient temperature in an anhydrous ether solvent such as diethyl ether, tetrahydrofuran and the like to produce corresponding l-(l-halo-3,4- dihydronaphth-2-yl)-alkan-l-ol which was treated with dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tribromide mixture at 0-10 0C for 20-60 hrs, contents neutralised with dilute alkali solution to produce 3-(l-halo-3,4-dihydronaphth- 2-yl)-2-alkyl-propenal. The 3-(l-halo-3,4-dihydronaphth-2-yl)-2-alkyl-propenal was condensed with an ylide (Wittig reagent, prepared from triphenylphosphine and methyl/ethyl bromoacetate or methyl/ethyl chloroacetate in equimolar. mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at temperature 5-80 0C for 6-60 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like or benzene/toluene or dimethylformamide to yield corresponding 5-(l-halo-3,4-dihydronaphth- 2-yl)-4-alkyl-2E,4E- pentadienoic acid ethyl ester which was saponified with a strong alkali solution using sodium hydroxide or potassium hydroxide followed by acidification to furnish 5-(l-halo-3,4-dihydronaphth-2-yl)-4-alkyl-2J?,4E-pentadienoic acid and amidation with acyclic or cyclic or heterocyclic amine to afford compounds of formula Ib. For the preparation of compound of formula lba, the acid of formula Ib was converted to its tetrohydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and 1-5 atmospheric pressure to obtain corresponding tetrohydro derivative which Was converted to corresponding amides of formula lba by the method described above for the preparation of compound of formula Ib. Alternatively lba may also be prepared by hydrogenation of compound of formula Ib as such by the method as described above. And excess of thionyl chloride and solvent removed. The acid chloride intermediate thus obtained was condensed with acyclic or cyclic or heterocyclic amine or aryl amine or C-protected amino ester or unsubstituted and substituted allcylaryl amines.
Method-4
The α-tetralone was first converted to 1,2,3,4-tetrahydronapth-l-ol by reduction with sodium borohydride and the resulting alcohol reacted with dimethylformamide and phosphorus oxybromide or phosphorus tribromide mixture at 0-10 0C for 5-24 hrs, contents neutralised with dilute alkali solution to produce 3,4-dihydronaphthalene-2-carbaldehyde. The 3,4-dihydronaphthalene-2-carbaldehyde was reacted with malonic acid in presence of base such as piperidine using pyridine as solvent, and the reaction left at room temperature for 24-30 hrs after that heated at 80-90 0C, contents were cooled and poured into ice cold water than acidified and extracted with ethyl acetate, organic layer was washed with water and dried over anhydrous sodium sulfate, concentrate furnished 3-(3,4-dihydro-naphth-2- yl)-acrylic acid. The resulting acid above is taken in an inert organic solvent such as benzene, dichloromethane and treated with thionyl chloride in inert organic solvent such as dichloromethane or benzene and the like to give compounds of the formula Ia or alternately 3,4-dihydronaphthalene-2-carbaldehyde was reacted with alkyl magnesium halide (alkyl halide C2-CiO, straight or branched) with constant stirring at ambient temperature in an anhydrous ether solvent such as diethyl ether, tetrahydrofuran and the like to produce corresponding l-(3,4-dihydronaphth-2-yl)-alkan-l-ol which was treated with dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tribromide mixture at 0-10 0C for 20-60 hrs, contents neutralised with dilute alkali solution to produce 3-(3,4-dihydronaphth-2-yl)-2-alkyl-propenal. The 3-(3,4-dihydronaphth-2-yl)~2- alkyl-propenal was condensed with an ylide (Wittig reagent, prepared from triphenylphosphine and methyl/ethyl bromoacetate or methyl/ethyl chloroacetate) in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at temperature 5-80 0C for 6-60 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like or benzene/toluene or dimethylformamide to yield corresponding 5-(3,4-dihydronaphth~2-yl)-4-alkyl-2£,4is-pentadienoic acid ethyl ester which was saponified with a strong alkali solution using sodium hydroxide or potassium hydroxide followed by acidification to furnish 5-(3,4-dihydronaphth-2-yl)-4-alkyl-2j5l,4£- pentadienoic acid which on treatment with acyclic or cyclic or heterocyclic amine aryl amine or C-protected amino ester or unsubstituted and substituted alkylaryl amines to afford compounds of formula Ib. For the preparation of compound of formula lba, the acid of formula Ib is converted to its tetrohydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and 1-5 5 atmospheric pressure to obtain corresponding tetrahydro derivative which was converted to corresponding amides of formula lba by the method described above for the preparation of compound of formula Ib. Alternatively lba may also be prepared by hydrogenation of compound of formula Ib as such by the method as described above.
Method-5
10 The alpha-tetraione was first allowed to react with Grignard reagent (prepared from aryl halides & magnesium or unsubstituted or substituted- benzyl halide) with constant stirring at ambient temperature in an anhydrous ether solvent such as diethyl ether, tetrahydrofuran and the like to produce corresponding phenyl (unsubstituted or substituted) dihydronaphth- 2-yl-carbinol or benzyl (unsubstituted or substituted) dihydronaphthyl carbinol which was
[5 treated with dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tribromide mixture at 0-10 0C for 20-60 hrs, contents neutralised with dilute alkali solution to produce 3-(3,4-dihydronaphth-2-yl)-2-aryl-propenal or 3-(3,4- dihydronaphth-2-yl)-2-benzyl-propenal. The propenal thus prepared was condensed with an ylide (Wittig reagent, prepared from tiϊphenylphosphine and methyl/ethyl bromoacetate or
10 methyl/ethyl chloroacetate) in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at temperature 5-80 0C for 6-60 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like or benzene/toluene or dimethylformamide to yield corresponding 5-(3,4-dihydronaphth-2-yl)-4-arylΛenzyl- 2E?4E-pentadienoic acid ethyl ester which was saponified with a strong alkali solution
15 using sodium hydroxide or potassium hydroxide followed by acidification to furnish 5-(3,4- dihydronaphth-2-yl)-4-aryl or benzyl-2E, 4£-pentadienoic acid which on treatment .with acyclic or cyclic or heterocyclic amine aryl amine or C-protected amino ester or unsubstituted and substituted alkylaryl amines to afford compounds of formula Ib. For the preparation of compound of formula lba, the acid of formula Ib is converted to its i0 hexahydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and 1-5 atmospheric pressure to obtain corresponding tetrohydro derivative which was converted to corresponding amides of formula lba by the method described above for the preparation of compound of formula Ib. Alternatively lba may also be prepared by hydrogenation of compound of formula Ib as such by the method as described above.
Schematic diagram
Figure imgf000023_0001
1ba
R=halogen, hydrogen, alkyl (Cl to ClO), substituted/unsubstituted aryl or benzyl. RlR2R3R4=hydrogen, nitro, halo, hydroxy, alkyloxy, alkenyloxy, aryloxy, substitutedpyranyl radical unsubstituted pyranyl radical or Rl , R4 represnet hydrogen and R2, R3 (R2=R3) represent hydrogen, hyroxyl, halo, nitro, alkoxyl' (Cl-ClO), alkenoxyl(Cl- ClO), aroyloxyl , substituted aroyloxyl , or Rl , R4 represnet hydrogen and R2+R3 together represent 1,3-dioxol or 1,4-dioxol ;and R5=R6= hydrogen or normal or branched chain Ci to C1O alkyl group or phenyl or benzyl radical or R5+R6=heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl, N- methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl etc. R7=normal or branched chain Cl to ClO alkyl group, phenyl, benzyl radical
Figure imgf000024_0001
R2=R3=H,OR(R=C1 Io C10, Aryl, Substituted aryl, Benzyl, Substituted benzyl x = H, Cl. Br, Phenyl, Substituted Phenyl, Benzyl, Substituted Benzyl, alkyl
R2+R3=1 ,3-dioxal, 1,4-dioxal
R5=R6=H, normal αr branched chain C1 to C10 alkyl group or phenyl or benzyl radical
R5+R6=heterocyc!ic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-melhylplperazlnyl, oxazolyl, N-methylpipera∑inyl, pyrrolyl, imidazolyl, oxazolyl, thiooxazole, an amino acid such as aianlnyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and Ihβ like and optionally converting them to (heir salts by method known In the art of synthesis,
R7= normal or branched chain C1 to C10 alkyl groups or phenyl or benzyl radical
Figure imgf000025_0001
branched chain) (substituted or unsubstituted)
Figure imgf000025_0002
yUnsubstltufcd)
Figure imgf000025_0003
Figure imgf000025_0004
(R=CH3, C1-C10 straight or branched chain)
Figure imgf000025_0005
or R=aryl or benzyl (substituted or unsubstituted)
Figure imgf000025_0006
Figure imgf000026_0001
(R=CH3, C1-C10 straight or R=halo,alkyl,aryl, benzyl branched chain)
Figure imgf000026_0002
X=CI,Br,alkyl,aryl,benzyl X=CI,Br,alkyl,aryl,benzyl
R=alky!,aryl, benzyl R=alkyl,aryl,benzyl
Bio efficacy New antibiotics are continued to be discovered at a rate of >500 per year, but these almost invariably belong to previously identified classes of compounds, making it likely that pathogens will be able to rapidly build up resistance to these "new" drugs. The increasing threat of drug-resistant pathogens is causing a renewed interest in the discovery of novel antibiotics. Present invention is focused on 'potentiators' involved wherein such agents, which by themselves are not therapeutic entities but when combined with an active drug, lead to the potentiation of the pharmacological effect of the drug. Such formulations/ combinations have been found to increase the bio-efficacy of a number of drugs even when reduced doses of drugs are present in such formulations. For example, patent nos. IP 172690; IP 176433 . and US 5744161 disclose such art. These compounds are not only capable of increasing bioavailability of a wide variety of therapeutic agents but are also capable of enhancing bio- efficacy through a variety of mechanisms. As a result newer understanding has emerged about the factors involved in decreased cellular concentrations of drugs at which they failed to attain therapeutic levels and the strategies that make it possible to enhance the bio- efficacy of these active drugs even at lower concentrations compared to standard high dosing. Some of these factors are:
(a) Increasing the penetration or entry of the active drug into the pathogen even where tney become persistors, besides inhibiting the capability of pathogens and abnormal tissues to reject the drug. This would eventually ensure the enhanced killing of the pathogenic microorganisms, which are otherwise inaccessible to the active drug.
(b) Chemo resistance is a major problem in drug therapy. The mechanisms underlying the clinical phenomena of de novo and acquired drug resistance may arise from alterations at any step in the cell-killing pathway. These include drug transport, drug metabolism, drug targets, cellular repair mechanisms and the ability of cells to recognize a harmful toxin or pathogen. A common mechanism of reduced cellular drug accumulation is the increased expression Multi ■ Drug Resistance pumps (MDRs). The present invention also proves that the compounds described here inhibit the bacterial efflux, as a result of which there is more of accumulation and decreased efflux of ethidium bromide is used as substrate to gauge the efflux of the drugs by the extent of fluorescence observed.
(c) Immunological intervention through nitric oxide (NO) production, CMI and/or humoral immune potentiation with favourable influence on the Th 1/ Th 2 balance.
(d) Sensitization of specific receptors like proteins, DNA, RNA etc thus potentiating and prolonging the effect leading to enhanced antibiotic activity against pathogens, and disorders. Adequate experimental evidences have been gained in respect of several of these mechanisms, (B. Ray, S. Medda, S. Mukhopadhyay, M.K. Basu, Indian J. Biochem. Biophys. 36 (1999) 248-251) modulating the membrane fluidity, which may alter the activity of membrane, bound transporter proteins. The overall permeability changes may affect (i) specific ion transporter channels, and (ii) also lead to bulk movement of lipophilic solutes along the paracellular pathway. Such membrane changes have also been evidenced in the action of several polyene antibiotics (J. Milhaud, J. Bolard, P. Benveniste, M.A. Hartmann, Biochim.
. Biophys. Acta. 943 (1988) 315-325).
(e) Potentiating the mechanism of action of drugs and thus increasing their efficacy at lower doses e.g. inhibition of RNA polymerase transcription leading to potentiation of the effect of rifampicin at less than half the standard dose. The products of the present invention are novel mechanism based pharmaceutical entities acting through synergism and/or additive effect so that drugs contained in the tormulation are more bio-efficaceous as a result of one or more of the mechanism as revealed above and thereby increasing the sensitivity of the target cell to an anti-infeetive.
Description of the preferred embodiment The Mrug' in the present invention refers to a chemical entity capable of affecting organism's patho-physiology and used for the treatment or prevention of disease. Drugs include a number of classes of compounds, but not limited to aminoglycoside, penicillins, cephalosporins and other β-lactam agents, macrolides, glycopeptides, fluoroquinolones, tetracyclines, first and second line anti-TB drugs, anti-leprosy, antiviral, polyene, triazole, and imidazoles and combinations like pyrimidines, sulphamethoxazole. Drugs may be a prodrug, activated or metabolised form, consisting of charged, uncharged, hydrophilic, hydrophobic or zwitter-ion species which make their entry by simple diffusion, carrier mediated transport dependent and not dependent on energy requirements, through ion and/ or voltage gated channels. The 'potentiator' refers to aromatic amides of the formulae Ia, laa, Ib, lba selected from the following set of compounds prepared by the method described in the examples. 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid piperidide 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid pyrrolidide 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid morpholide 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid n-octylamide
3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid isopropylamide 3-(l -chloro-3,4-dihydfonapth-2-yl)-acrylic acid N,N-diethylamide 3 -(I -chloro-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisopropylamide 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acidp-methoxyanilide
3 -(I -chloro-3,4-dihydronapth-2-yl)-acrylic acid 3,4-methylenedioxyanilide 3-(l -chloro-3,4-dihydro-napth-2-yl)-acrylic acid o-methylanilide 3-(l -chloro-3,4-dihydronapth-2-yl)-acrylic acid benzylamide 3-(l -chloro-3,4-dihydronapth-2-yl)-acrylic acid p-nitroanilide 3-(l -chloro-3,4-dihydro-napth-2-yl)-acrylic acid p-fluoroanilide
3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acidp-carboxyanilide 3 -(I -chloro-3,4-dihydronapth-2-yl)-acrylic acid N,N-diphenylamide 3 -(I -chloro-3,4-dihydronapth-2-yl)-acrylic acid N-methylpiperazide 3 -( 1 -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid />-methylanilide 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid «-butylamide 5 3 -(I -chloro-3 ,4-dihydronaρth-2-yl)-acrylic acid anilide
3-(l -chloro-3, 4-dihydronapth-2-yl)-acrylic acid imidazolide 3 -( 1 -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid ø-methoxyanilide 3 -( 1 -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid N,N-diisobutylamide '3-(l-bromo-3,4-dihydiOnapth-2-yl)-acrylic acid piperidide
LO 3 -( 1 -bromo-3 ,4-dihydronapth-2-yl)-acrylic acid pyrrolidide
3 -(I -bromo-3, 4-dihydronapth-2-yl)-acrylic acid morpholide 3 -(I -bromo-3 ,4-dihydronapth-2-yl)-acrylic acid isobutylamide 3-(l -bromo-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisopropylamide 3-(l-bromo-3,4-dihydro-napth-2-yl)-acrylic acid «-butylamide
15 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid N-methylpiperazide
3-( 1 -bromo-3 ,4~dihydronapth-2-yl)-acrylic acid N,N-diethylamide 3-(l -bromo-3 ,4-dihydro-napth-2-yl)-acrylic acid imidazolide 3-(l -bromo-3 ,4-dihydronapth-2-yl)-acrylic acid />-hydroxypiperidide 3-(3,4-dihydronaphth-2-yl)-acrylic acid isobutylamide
'0 3-(3,4-dihydronaphth-2-yl)-acrylic acid N,N-diisopropylamide '
3-(3.4-dihydronaphth-2-yl)-acrylic acid piperidide 3-(3,4-dihydronaphth-2-yl)-acrylic acid pyrrolidide 3-(3,4-dihydronaphth-2-yl)-acrylic acid morpholidide 3-(3,4-dihydronaphth-2-yl)-acrylic acid/>-methoxy anilide
:5 3-(3,4-dihydronaphth-2-yl)-acrylic acid «-octylamide
3-(3,4-dihydronaphth-2-yl)-acrylic acid N,N-diethylamide 3-(3,4-dihydronaphth-2-yl)-acrylic acid anilide 3-(3,4-dihydronaphth-2-yl)-acrylic acid 3,4-methylenedioxy anilide 3-(l-benzyl-3,4-dihydro-naρth-2-yl)-acrylic acid «-octylamide
0 3-(l-benzyl-3,4-dihydro-napth-2-yl)-acrylic acid piperidide
3-(l-benzyl-3,4-dihydro-napth-2-yl)-acrylic acid pyrrolidide . 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid morpholide 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisopropylamide 3 -( 1 -benzyl-3 ,4-dihydronapth-2-yl)-acrylic acid ^-hydroxy piperidine 5 3 -(6,7-dimethoxy-3 ,4-dihydronaphth-2-yl)-ρropenoic acid piperidide
3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-ρropenoic acid moφholide 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid pyrrolidide 3 -(6,7-dimethoxy-3 ,4-dihydronaphth-2-yl)-propenoic acid n-octylamide 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid isobutylamide
10 3 -(6,7-dimethoxy-3,4-dihydronaρhth-2-yl)-ρropenoic acid rø-butylamϊde
3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid n-propylamide 3 -(6,7-dimethoxy-3 ,4-dihydronaρhth-2-yl)-propenoic acid isopropylamide 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid N,N-diisopropylamide 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid anilide
15 3-(6,7-diraethoxy-3,4-dihydronaphth-2-yl)-piOpenoic-acid-o-methylanilide
3 -(6,7-dimethoxy-3 ,4-dihydronaphth-2-yl)-propenoic acid /7-raethylanilide 3 -(6,7-dimethoxy-3 ,4-dihydronaphth-2-yl)-propenoic acid /7-methoxyanilide 5-( 1 -chloro-3 ,4-dihydronaphth-2-yl)-4~methyl-2£,4i?-pentadienoic acid isobutylamide
-0 5-(l-chloro-3,4-dihydronaphth-2-yl)-4-methyl-2£',4£'-pentadienoic acid N5N- diisopropylamide
5-( 1 -chloro-3 ,4-dihydronaphth-2-yl)-4-methyl-2E,4E-pentadienoic acid piperidide 5-(l -chloiO-3,4-dihydronaphtli-2-yl)-4-methyl-2E,4£-pentadienoic acid pyrrolidide 5-(l-chloro-3.4-dihydronaphth-2-yl)-4-methyl-2£,4£'-pentadienoic acid morpholide
!5 5-(l -chloro-3, 4-dihydronaphth-2-yl)-4-ethyl-2£',4^'-ρentadienoic acid piperidide
5-(l-chloro-3,4-dihydronaphth-2-yl)-4-ethyl-2£,4£-pentadienoic acid pyrrolidide 5-( 1 -chloro-3 ,4-dihydronaphth-2-yl)-4-ethyl-2E,4£'-ρentadienoic acid isobutylamide 5~(l-chloro-3,4-dihydronaphth-2-yl)-4-ethyl-2£?4£'-pentadienoic acid N,N- diisopropylamide.
0 3 -( 1 -methyl-3 ,4-dihydronapth-2-yl)-acrylic acid piperidide
3-(l-methyl-3,4-dihydronapth-2-yl)-acrylic acid pyrrolidide 3-(l-methyl-3,4-dihydronapth-2-yl)-acrylic acid morpholide 3 -(I -methyl-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide 3-(l-methyl-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisopropylamide 3-(l -chloro- l,2,3,4-tetrahydiOnapth-2-yl)-propanoic acid piperidide 3-(l-chloro-l,2,3,4-tetrahydronapth-2-yl)-propanoic acid pyrrolidide
3-(l-chloro-l,2,3,4-tetrahydronapth-2-yl)-propanoic acid morpholide 3 -( 1 -chloro -1,2,3 ,4-tetrahydronaρth-2-y l)-propanoic acid n-buty lamide 3 -( 1 -chloro- 1 ,2,3 ,4-tetrahydiOnapth-2-yl)-propanoic acid isobutylamide 3-(l-chloro-l,2,3,4-tetrahydronapth-2-yl)-propanoic acid π-octylamide 3-(l-chloro-l,2,3,4-tetrahydronapth-2-yl)-propanoic acid N,N-diethylamide
3-(l -chloro-1 ,2,3 ,4-tetrahydronapth-2-yl)-propanoic acid N,N-diisopropylamide 3 -( 1 -bromo- 1 ,2,3 ,4-tetrahydronapth-2-yl)-propanoic acid piperidide 3-(l-bromo-l ,2,3,4-tetrahydronapth-2-yl)-propanoic acid isobutylamide 3-(l -bromo- 1 ,2,3 ,4-tetrahydronapth-2-yl)-propanoic acid ^-methoxyanilide 3-(l -bromo- 1 ,2,3,4-tetrahydronapth-2-yl)-propanoic acid N,N-diisopropylamide
3-(l-bromo-l,2,3,4-tetrahydronapth-2-yl)-propanoic acid pyrrolidide 3-(l -bromo- 1 ,2,3,4-tetrahydronapth-2-yl)-propanoic acid ø-methylanilide 3-(l-bromo-l, 2,3 ,4-tetrahydronapth-2-yl)-propanoic acid />-methylanilide 5-( 1 -chloro- 1 ,2,3 ,4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid N5N- diisopropylamide
5-(l-chloro-l,2,3,4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid isobutylamide 5-(l -chloro-1 ,2,3,4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid piperidide 5-(l-chloro-l,2,3,4-tetrohydronaphth-2-yl)-4-niethyl-pentanoic acid pyrrolidide 5-(l -chloro-1 ,2,3,4-tetrohydronaphth-2-yl)-4-methyl-ρentanoic acid n-octylamide 5-(l -chloro-1, 2,3 ,4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid anilide
5-(l-chloro-l ,2,3,4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid morpholide. 5-(l -chloro-1, 2,3, 4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid glycine methyl ester amide 5-(l-chloro-l,2,3,4-tetrohydronaphth-2-yl)-4-methyl-ρentanoic acid alanine methyl ester amide
5-(l -chloro-1, 2,3, 4-tetrohydronaphth-2-yl)-4-methyl-ρentanoic acid phenyl alanine methyl ester amide
5 -( 1 -chloro- 1,2.3 ,4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid triptamine amide.
The combination of particular amount of potentiator with particular amount of the drug i.e the doses of efflux pump inhibitor and the antimicrobial agent which are useful in combination as a treatment are therapeutically effective which generally refers to the inhibition of the normal metabolism of the micro-organisms responsible for microbial infection. In the present invention, a therapeutically effective amount means those amounts of the efflux pump inhibitor and anti-microbial agent which, when used in combination produce the desired therapeutic effect as judged by the model animal infection studies. In particular embodiment, the efflux pump inhibitor and anti-microbial agent are combined in predetermined proportions and thus a therapeutically effective amount would be an amount of the combination and the amount of individual components i.e. efflux pump inhibitor and the anti-microbial agent as well as the combination of two can be determined by one of the skill in the art and the amount of the combination will vary depending upon several factors such as the particular microbial strain /isolate used in the study and the particular efflux pump inhibitor and the anti-microbial agent used besides depending upon the weight, sex, age etc of the animal/patient on whom the combination of the efflux pump inhibitor and the anti-microbial agent is tested. A pharmaceutical carrier is generally an inert bulk agent added to make the ingredients achieve superior ad mixing and can be solid or liquid. The inert parts of standard pharmaceutical compositions used in this process are also part of the present invention.
Study design
The checker board method:
This is the most frequently used method to access the antimicrobial combinations in vitro. The term "checkerboard" refers to the pattern (of tubes or microtiter plate wells) formed by multiple dilutions of two drugs being tested (Eliopoulos GM, Moelle'ring RC. Antimicrobial Combinations, in: Antibiotics in Laboratory Medicine: USA: Williams & Willάns). In the present study the checkerboard consisted of columns in which each tube (or well) contains the same amount of the standard drug (antibacterial/antifungal/anti-TB/antiviral) being diluted along the x-axis and rows in which each tube (or well) contains the same amount of the potentiator being diluted on the y-axis. As a result each square in the checkerboard (which represents one tube/ well or plate) contained a unique. This chequerboard technique can be performed with liquid or semisolid (agar) media.
Agar Method:
In this method the agar (Mueller Hinton agar, Middlebrook 7H10 agar) was autoclaved and allowed to cool to 55 0C to 50 0C. The combination of the standard drug and the potentiator was added to the agar. Serial two fold dilutions of each of standard drug and the potentiator were prepared in appropriate solvents. In order to maintain the desired concentrations of both agar and drugs, and to rule out the effect of solvent, the volume of solvent (containing standard drug or potentiator) added to agar was kept small (i.e. 5% of the total volume). After the agar plates have been poured and allowed to dry, the bacteria to be tested were applied to the surface of agar with a replicating device designed to deliver a standard inoculum (approx 104 cfu|spot). The plates were incubated at 37 0C for 24 hrs (3 weeks in case of Mycobacterium tuberculosis).
-Broth Method: The above-mentioned checkerboard was also performed with liquid media in a microtiter plate format. This method was used to study the combination of antibacterial/antifungal/ antiviral drugs with potentiator.
Toxicity evaluation of 3-(l-chloro-3,4-dihydro-napth-2-yl)-acrylic acid N,N- diisopropylamide of formula Ia Toxicity evaluation of 3-(l-chloro-3,4-dihydro-napth-2-yl)-acrylic acid N,N- diisopropylamide of formula Ia (example-5). In acute toxicity studies an LD50 (oral) was found to be >3.0 gm/kg in mice. In subacute studies the effect of 3-(l-chloro-3,4-dihydro- napth-2-yl)-acrylic acid N,N-diisopropylamide of formula Ia (example-8) at 20, 40 and 100 mg/kg was investigated for 28 days. Parameters monitored were: food/water consumption, body weight, organ weight, haematological and clinical chemistry profile. No adverse effects were observed. Safety pharmacology studies (CNS, CVS and GIT profile) did not reveal any abnormality.
The following examples are intended to demonstrate some of the preferred embodiments but in no way be construed so as to limit the scope of the invention. Any person skilled in the art can design more formulations, which may be considered as part of the present invention.
Example 1
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yI)-acrylic acid piperidide of formula-l where
Figure imgf000034_0001
1) Preparation of l-chloro-2-formyl-3,4-dihydronaphthalene
To a chilled solution of α tetratone (2Og, 134 mmol) in dry DMF (50 ml), POCl3 (12.6 ml) was added drop wise at 0-5 0C for lhr and the resulting mixture was stirred at room temperature for 24 hrs. The reaction mixture was poured into ice-cold water (150 ml) containing sodium acetate (2Og). The contents were extracted with ethyl acetate (3x100 ml). The combined extracts were pooled and washed with water (3x50 ml), dried over anhydrous sodium sulfate and concentrated on rota vapour to give crude product, which was purified on silica gel column using pet. ether 60-80 0C: ethyl acetate (98:2) as an eluent to gave yellow viscous oil l-chloro-2-formyl-3,4-dihydronaphthalene yielded (24.8g, 95%), analysed for C11H9ClO; calcd C 68.58 H 4.71 Cl 18.40%, found C 69.04 H 4.72 Cl 18.41%) 2) Preparation of 3-(l-chloro-3,4-dihydronaphth-2-yI) acrylic acid
To a mixture of l-chloro-2-formyl-3,4-dihydronaphthalene (18.0g, 93 mmol) in pyridine (50 ml) and piperidine (5 ml), malonic acid (18g 1:2 eq) was added and the reaction mixture kept at room temperature for 52 hrs and heated on water bath for 6 hrs. To the cooled contents added 50% aqueous HCl solution and the resulting precipitate filtered, washed with water, dried and crystallized in pet.ether/ethylacetate to gave 3-(l-chloro-3,4-dihydronaphth- 2-yl) acrylic acid yielded (19.6g 90%), analysed for C13Hi1ClO2; calcd C 66.53 H 4.72 Cl 15.11%, found C 66.99 H 4.77 Cl 15.31%. 1H NMR (200 MHz, CDCl3): δ 2.63 (2H, t, J=7.82 Hz, CH2), 2.91 (2H, t, J=7.82 Hz, CH2), 6.11 (IH, d, J=15.80 Hz, CH=CHCO), 7.25 (4H, m, 4xAr-H), 8.25 (IH, d, J=I 5.83 Hz, CH=CHCO). 3) Preparation of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid piperidide._
To the compound 3-(l-chloro-3,4-dihydronaphth-2-yl) acrylic acid (500mg, 2.31mmol) in dry benzene (50 ml) added freshly distilled thionyl chloride (1.0 ml) and refluxed for lhr, excess of thionyl chloride removed in vacuo and thereafter condensed with dry benzene
5 solution of piperidine (1.0 ml) and stirred for 30 min. The organic layer washed with water (2x25 ml), dried over anhydrous sodium sulfate and evaporated on rotary evaporator to give crude product, which was purified on silica gel column using pet. ether 60-80 0C: ethyl acetate (90:10) as an elueht to gave the title compound (620mg, 94%), analysed for C18H20ClNO; calcd C 71.63 H 6.68 Cl 11.75 N 4.64%, found C 72.02 H 6.74 Cl 12.01 N
10- 4.71%. 1H NMR (200 MHz CDCl3): δ 1.62 (6H, bs, 3xCH2), 2.61 (2H, d, J=8.2Hz, CH2CH2), 2.88 (2H, d, J=8.2Hz, CH2CH2), 3.56 (4H, m, 2xCH2), 6.54 (IH, d J=I 5.43 Hz, CH=CHCO), 7.23 (3H, m, 3xAr-H), 7.71 (IH, dd, J=2.4, 9.0 Hz, Ar-H), 8.03 (IH, d, 15.44 Hz, CH=CHCO). Example 2
'.5 Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acryIic acid pyrrolidide of formula 1 where
Figure imgf000035_0001
R=Cl and R5+R6=pyrrolidine
This was similarly prepared, as described in example 1, except the starting materials were 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and pyrrolidine. Yield (90%), analysed for C18H20ClNO; calcd C 71.63 H 6.68 Cl l l!75 N 4.64%, found C71.99 H 6.82 Cl 12.11 N
10 4.66%. 1H NMR (CDCl3): δ 1.96 (4H, m), 2.66 (2H, t, J=5.0 Hz), 2.89 (2H, t, J=5.0 Hz), 3.51 (2H, t, J=6.5 Hz), 3.65 (2H, t, JN5.5 Hz), 6.57 (IH, d, J=15.36 Hz), 7.25 (3H, m), 7.70 (IH, d, >2.76 Hz), 8.06 (IH, d, J=15.3 Hz).
Example 3
5 Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yI)-acrylic acid morpholide of formula 1 where Ri=R2=Ra=Ri=H, R=Cl and R5+R6=morpholidine
This was similarly prepared, as described in example 1, except the starting materials were 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and morpholine. Yield (95%), analysed for C17H18ClNO2; calcd C 67.21 H 5.97 Cl 11.67 N 4.61%, found C 68.11 H 6.00 Cl 12.1 N 0 4.59%. 1H NMR (CDCl3): δ 2.68 (2H, t, J=8.4 Hz), 2.87 (2H, t, J=8.4 Hz), 3.31 (4H,- m), 3.73 (4H, m), 6.77 (IH, d, J=15.3 Hz), 7.26 (3H, m), 7.69 (IH, d, J=5.5 Hz), 8.07 (IH, d, J=I 5.3 Hz)
Example 4
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid λi-octylamide of formula 1 5 where
Figure imgf000036_0001
This was similarly prepared, as described in example I3 except the starting materials were 3-(l-chloro-3,4-dihydronapth-2-yl)-acryIic acid and /i-octylamine. Yield (73%), analysed for C2iH28CINO; calcd C 72.92 H 8.16 CI 10.25 N 4.05%, found C 73.11 H 8.22 Cl 10.44 N 4.10%. 1HNMR (CDCl3): δ 0.87 (3H, t, /=6.68 Hz), 1.28 (12H, m), 1.55 10 (2H, t, /=7.16 Hz), 2.61 (2H, t, /=5.20 Hz), 2.87 (2H, t, /=5.14 Hz), 6.30 (IH, d, /=15.5 Hz), 7.24 (3H, m), 7.69 (IH, d, /=2.23 Hz), 8.0 (IH, d, /=15.5Hz).
Example 5
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide of formula 1 15 where
Figure imgf000036_0002
R=CI and R5+R6=isobutylamine.
This was also similarly prepared, as described in example 1, except the starting materials were 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and isobutylamine. Yield (73%), analysed for Ci7H20ClNO; calcd C 70.46 H 6.96 Cl 12.23 N 4.83%, found C 70.61 H 7.01 Cl 12.61 N 4.88 %. 1HNMR (200 MHz CDCl3): δ 0.94 (6H, d, J=6>7 Hz, CH (CRi)2), 1-83 >0 (IH3 m, CH(CH3),), 2.62 (2H, t, J=7.75 Hz5CH2-CH2), 2.88 (2H, t, J=7.76 Hz, CH2CH2), 3.11 (2H, d, J=6.4 Hz, CH2CH), 6.33 (IH, d, J=15.54 Hz, CH=CHCO), 7.25 (3H, m, 3xAr- H), 7.69 (IH, m, Ar-H), 8.01 (IH, d, J=I 5.57 Hz, CH=CHCO).
Example 6
\5 Synthesis of 3-(l-chloro-3,4-dihydroπapth-2-yI)-acrylic acid isopropylamide of formula 1 where Ri=R2=R3=Ri=H, R=CI and R5+R6=isopropylamine.
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and isopropyl amine. Yield (90%), analysed for Ci6Hi8ClNO; calcd C 69.68 H 6.58 Cl 12.86 N 5.08%, found C 70.02 H 6.66 Cl 13.11 N 0 5.11%. 1H NMR (CDCl3): δ 1.19 (6H, d, J=6.58 Hz), 2.61 (2H, t, J=8.3 Hz), 2.88 (2H, t, J=8.3 Hz), 4.07 (IH, m), 6.28 (IH, d, J=6.28 Hz), 7.24 (3H, m), 7.69 (IH, d, J=4.5 Hz), 8.00 (IH5 d, J=15.6 Hz).
Example 7
Synthesis of 3-(l-chIoro-3,4-dihydronapth-2-yl)-acrylic acid-N,N-diethylamide of 5 formula 1 where Ri=R2=Ra=RpH, R=CI and Rs+R^JN^N-diethylamine.
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and N,N-diethylamme. Yield (85%), analysed for C2ιH28ClNO; calcd C 72.92 H 8.16 Cl 10.25 N 4.05%, found C 73.11 H 8.22 Cl 10.55 N 4.11%. 1HNMR (CDCl3): δ 1.20 (6H5 m), 2.65 (2H, t, J=5.24 Hz), 2.88 (2H, t, J=5.O3 Hz), 10 3.49 (4H5 m), 6.67 (IH, d, J=15.28 Hz)5 7.23 (3H5 m), 7.69 (IH, d, J=2.23 Hz), 8.07 (IH, d, J=I 5.28 Hz).
Example 8
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid-N,N-diisopropylamide of 15 formula 1 where Ri=R2=R3=R4=H, R=Cl and Rs+R6=N,N-diisopropylamine.
This was similarly prepared, as described in example I5 except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and. diisopropylamine Yield (98%), analysed for C19H24ClNO; calcd C 71.80 H 7.61 Cl 1 1.15 N 4.41%, found C 72.61 H 7.69 Cl 11.23 N 4.48 %. 1H NMR (200 MHz CDCl3): δ 1.23 & 1.27 (6H each, d, J=6.7 Hz each, 2xCH .0 (CH3)2), 1.96 (2H, m, 2xCH(CH3)2), 2.70 (2H, d, J=7.45 Hz, CH2CH.), 2.89 (2H, d, J=7.45 Hz, CH2CH2), 6.80 (IH, d, J=15.54 Hz, CH=CHCO), 7.30 (3H, m, 3xAr-H), 7.70 (IH, m, Ar-H), 7.97 (IH, d, 15.57 Hz, CH=CHCO).
Example 9 i5 Synthesis of 3-(l-chIoro-3,4-dihydronapth-2-yl)-acryIic acid />-methoxyanilide of formula 1 where
Figure imgf000037_0001
This was similarly prepared, as described in examplel, except the starting materials were 3- (l-chloro-3,4-dih3^dronapth-2-yl)-acrylic acid and jσ-anisidine. Yield (89%), analysed for C20H18ClNO2; calcd C 70.69 H 5.34 Cl 10.34 N 4.12%, found C 71.43 H 5.39 Cl 1039 N ;0 4.26%. 1H NMR (200 MHz CDCl3): δ 2.64 (2H, t, J=7.6 Hz)5 2.91- (2H, t, J=7.6 Hz), 3.73 (3H, s), 6.55 (IH5 d, J=15.40 Hz), 2.91 (2H, d, J=8.97 Hz), 7.33 (2H5 d5 J=6.9 Hz), 7.64 (4H5 m), 7.99 (IH, d, J=15.4 Hz), 10.15 (IH, s).
Example 10
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acryIic acid-3,4-methylenedioxy- 5 anilide of formula 1 where
Figure imgf000038_0001
R=Cl and R3+R6=3,4-methylenedioxy- aniline.
This was similarly prepared, as described in examplel, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acryiic acid and 3,4-methylenedioxy aniline. Yield (84%), analysed for C20Hi6ClNO3; calcd C 67.90 H 4.56 Cl 10.02 N 3.96%, found C 68.64 H 5.59 10 Cl 10.14 N 4.03%.
Example 11
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acryIic acid ø-methylanilide of formula
1 where
Figure imgf000038_0002
15 This was similarly prepared, as described in example 1, except the starting materials were 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and ø-methylaniline. Yield (90%), analysed for C20HisClNO; calcd C 74.18 H 5.60 Cl 10.95 N 4.33%, found C .74.26 H 5.82 Cl 11.03 N 4.39%.
>0 Example 12
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acryIic acid benzylamide of formula 1 where
Figure imgf000038_0003
R=Cl and Rs+R6=benzylamine.
This was similarly prepared, as described in example 1, except the starting materials were 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and benzylamine. Yield (73%), analysed for 15 C20Hi8ClNO; calcd C 74.18 H 5.60 Cl 10.95 N 4.33%, found C 74.26 H 5.79 Cl 10.99 N 4.38%.
Example 13
Synthesis of 3-(l-chIoro-3,4-dihydronapth-2-yl)-acrylic acid />-nitroaniIide of formula 1 0 where
Figure imgf000038_0004
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and p-nitroaniline. Yield (73%), analysed for C19H15ClN2O3; calcd C 64.32 H 4.26 Cl 9.99 N 7.90%, found C 64.92 H 4.31 Cl 10.07 N 7.99%.
5 Example 14
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acryIic acid p-fluoroanilide of formula 1 where
Figure imgf000039_0001
R=Cl and
Figure imgf000039_0002
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and p-fiuoro aniline. Yield (71%), analysed 10 for Ci9Hi5ClFNO; calcd C 69.62 H 4.61 Cl 10.82 F 5.80 N 4.27%, found C 70.44 H 4.69 Cl 10.90 N 4.32%.
Example 15
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid />-carboxyaniIide of 15 formula 1 where
Figure imgf000039_0003
This was similarly prepared, as described in example 1, except the starting materials were
3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and p-carboxyaniline. Yield (73%), analysed for Ci9Hi6ClNO3; calcd C 67.90 H 4.56 Cl 10.02 N 3.96%, found C 68.80 H 4.60
Cl 10.09 N 3.99%. 20
Example 16
Synthesis of 3-(l-chloro-3, 4-dihydronapth-2-yl)-acrylic acid N,N-diphenylamine of formula 1 where Ri-R2=Rs=Ri=H, R=Cl and R5+R6=N,N-diphenylamine.
This was similarly prepared, as described in example 1, except the starting materials were .5 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and diphenylamine. Yield (83%), analysed for C25H20ClNO; calcd C 77.81 H 5.22 Cl 9.19 N 3.63%, found C 78.69 H 5.29 Cl 9,27 N
3.68%.
Example 17
»0 Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid N-methylpiperazide of formula 1 where
Figure imgf000039_0004
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and N-methylpiperazine. Yield (85%), analysed for C18H2iClN2O; calcd C 68.24 H 6.88 Cl 11.19 N 8.84%, found C 69.16 H 6.89 Cl 11.24 N 8.91%.
Example 18
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acryIic acid p-methylanilide of formula 1 where R1=R2=R3=R4=H, R=Cl and R5+R<;=/;-methyIaniline.
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and j9-methylaniline. Yield (90%), analysed for C20H18ClNO; calcd C 74.18 H 5.60 Cl 10.95 N 4.33%, found C 75.10 H 5.66 Cl 10.99 N 4.39%.
Example 19 Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yI)-acryIic acid /ι-butylamide of formula 1 where Ri=R2=R3=Ri=H, R=Cl and Rs+R6=«-butylamine.
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and π-butylamine. Yield (90%), analysed for C17H20ClNO; calcd C 70.46 H 6.96 Cl 12.23 N 4.83%, found C 71.49 H 7.00 Cl 12.27 N 4.89%.
Example 20
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid anilide of formula 1 where
Figure imgf000040_0001
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and aniline. Yield (94%), analysed for Ci9H16ClNO; calcd C 73.66 H 5.21 Cl 11.44, N 4.52%, found C 74.54 H 5.26 Cl 11.47 N 4.59%.
Example 21
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid imidazolide of formula 1 where R1=R2=R3=Rt=H, R=Cl and R5+Rδ=imidazole.
This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and imidazole. Yield (94%), analysed for C16H17C1N2O; found C 66.55 H 5.93 Cl 12.98 N 9.70%, calcd C 66.94 H 5.99 Cl 13.02 N 5 9.77%.
Example 22
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid ø-methoxyanilide of formula 1 where Ri=R2=R3=R4=H, R=Cl and Rs+R^ø-methoxyaniline.
10 This was similarly prepared, as described in example 1, except the starting materials were 3- (l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and ø-methoxyaniline. Yield (90%), analysed for C20H18ClNO2; calcd C 70.68 H 5.34 Cl 10.43 N 4.12%, Found C 71.61 H 5.39 Cl 10.38
N 4.19%.
L 5 Example 23
Synthesis of 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisobutylamide of formula 1 where Ri=R2=R3=R-J=H, R=Cl and R3+R6=N,N-diisobutylamine.
This was similarly prepared, as described in example 1, except the starting materials were 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid and N5N-diisobutylamine. Yield (94%), >0 analysed for C21H28ClNO; calcd C 72.92 H 8.16 Cl 10.25 N 4.05%, found C 73.88 H 8.21 Cl 10.30 N 4.14%.
Example 24
Synthesis of 3-(l-bromo-3, 4-dihydronapth-2-yl)-acrylic acid piperidide of formula 1 15 where
Figure imgf000041_0001
R=Br and Rg+R6=piperidine.
1) Preparation of l-bromo-2-formyl-3,4-dihydronaphthalene
Dry DMF (8.02 ml, 103.2 mmol) in dry dichloromethane (DCM) (50 ml) was cooled to 0° C and phosphorus tribromide (8.0 ml, 89.5 mmol) was added drop-wise. The mixture was stirred at 0° C for 1 hr during which time a pale yellow suspension was formed. A solution 0 of -tetralone (5,03g, 34.4 mmol) in dry DCM (90 ml) was added and the mixture was heated under reflux for 1 hr. After cooling to 0 0C, aqueous NaHCO3 was added slowly until the effervescence had subsided. Extraction of the reaction mixture with DCM (3x100ml) was followed by drying the organic layer (MgSO4). The solution was filtered through a celite plug and evaporation of the excess solvent resulted into brown oil. Column chromatography eluting with ethyl acetate :hexane (20:80) gave the product as a brown solid l-bromo-2- formyl-3,4-dihydronaphthalene (6.7Og, 80%), analysed for CnH9BrO; calcd C 55.72 H 3.83 Br 33.70%, found C 56.69 H 3.89 Br 33.77%. 1H NMR (CDCl3): 2.61 (2H, t, J=8'.l Hz)5 3.95 (2H51, J-8.1 Hz)5 7.28 (3H5 m), 7.89 (IH5 d, J=7.2 Hz)5 10.25 (IH5 s).
2) Preparation of 3-(l-bromo-3,4-dihydro-napth-2-yl)-acrylic acid
To a mixture of l-bromo-2-formyl-354-dihydronaphthalene (5.Og5 21 mmol) in pyridine (15 ml) and piperidine (2.5 ml), malonic acid (2.63g5 1.2 eq) was added and kept the reaction mixture at room temperature for 52 hrs. Later the reaction mixture was heated on water bath for 6 hrs, and poured the contents in to a conical flask. To this 50% aqueous HCl solution was added to precipitate the 3-(l-bromo-3,4-dihydro-napth-2-yl)-acrylic acid. This precipitate was filtered and crystallized in pet. ether/ethyl acetate to yield pure product 3 (5.56g, 95%), analysed for C13HnBrO2; calcd C 55.94 H 3.97 Br 28.68%, found C 56.77 H
4.02 Br 28.72%. 1H NMR (CDCl3): δ 2.61 (2H5 1, 8.2 Hz)5 2.80 (2H5 1, J=8,2 Hz), 6.13 (IH5 d, J=15.7 Hz), 7.32 (3H, m), 7.78 (IH, d, J=4.9 Hz)5 8.21 (IH5 d, J=I 5.7 Hz).
3) Preparation of 3-(l-bromo-3,4-dihydronapth-2-yI)-acrylic acid piperidide._
To the compound 3-(l-bromo-354-dihydro-napth-2-yl)-acrylic acid (500mg, 2.31mmol) in dry benzene (15 ml) added freshly distilled thionyl chloride (0.53 ml) and refluxed for lhr, excess of thionyl chloride removed in vacuo and thereafter condensed with dry benzene solution of piperidine (1.0 ml) and stirred for 30 min. The organic layer was washed with water (2x25 ml), dried over anhydrous sodium sulfate, concentrated to give crude product, which was purified on silica gel column using pet. ether: ethyl acetate (92:8) as an eluent to gave the title compound (620mg, 98%), analysed for C18H20BrNO; calcd C 62.44 H 5.82 Br
23.08 N 4.05%, found C 63.47 H 5.91 Br 23.13 N 4.11%. 1H NMR (200 MHz CDCl3): δ
0.96 (6H5 d, J=6.6 Hz5 CH (CH3)2)5 1.83 (IH, m, CH(CH3)2), 2.57 (2H5 t, J=7.75
Hz,CH2.CH2)5 2.87 (2H, t, J=7.76 Hz, CH2CH2), 3.20 (2H, d, J=6.4 Hz, CH2CH), 6.11 (IH5 d, J=15.47 Hz, CH=CHCO), 7.21 (3Η, m5 3xAr-H), 7.75 (IH, dd, J=2.0, 8.73 Hz, Ar-H), 8÷01 (IH, d, J=15.47 Hz, CH=CHCO). Example 25
Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid pyrrolidide of formula 1 where Ri=R2=R3=R4=H, R=Br and R5+R6=pyrrolidine.
This was similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid and pyrrolidine. Yield (90%), analysed for C17H18BrNO; calcd C 61.46 H 5.46 Br 24.05 N 4.22%, found C 62.41 H 5.49 Br 24.08 N 4.36%.
Example 26 Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid morpholide of formula 1 where RI=R2=RJ=RJ=H, R=Br and Rs+Rβ =morpholine.
This was similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid and morpholine. Yield (95%), analysed for C17Hi8BrNO2; calcd C 58.63 H 5.21 Br 22.95 N 4.02%, found C 59.41 H 5.29 Br 23.08 N 4.09%
Example 27
Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide of formula
1 where
Figure imgf000043_0001
This was similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid and isobutylamine. Yield (92%), analysed for Ci7H20BrNO; calcd C 61.09 H 6.03 Br 23.91 N 4.79%, found C 61.99 H 6.11 Br 24.02 N 4.83%. 1H NMR (CDCl3): δ 0.95 (6H, d, J=6.66 Hz)5 1.83 (IH, m), 2.57 (2H, t, J=8,2 Hz), 2.87 (2H, t, J=8.2 Hz), 3.21 (2H, d, J=6.60 Hz), 6.11 (IH, d, J=15.5 Hz), 7.21 (3H, m), 7.75 (IH3 d, J=4.5 Hz), 8.00 (IH, d, J=15.5 Hz).
Example 28
Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisopropyIamide of formula 1 where
Figure imgf000043_0002
This was similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid and N,N-diisopropylamine. Yield (92%), analysed for C19H24BrNO; calcd C 62.99 H 6.68 Br 22.05 N 3.87%, found C 63.44 H 6.71 Br 22.09 N 3.91%.
Example 29 Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acryIic acid n-butylamide of formula 1 where
Figure imgf000044_0001
This was similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid and n-butyl amine. Yield (86%), analysed for C17H20BrNO; calcd C 61.09 H 6.03 Br 23.96 N 4.19%, found C 61.97 H 6.11 Br 24.04 N 4.24%.
Example 30
Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acryIic acid N-methylpiperizide of formula 1 where Ri=R2=Ra=Ri=H, R=Br and Rs+R6=N-methyIpiperazine. This was similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid and N-methylpiperizine. Yield (86%), analysed for C18H2iBrN2O; calcd C 59.84 H 5.86 Br 22.12 N 7.755, found C 60.77 H 5.91 Br 22.19 N 7.84%.
Example 31
Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid N,N-diethylamide of formula 1 where Ri=R2=Ra=Ri=H, R=Br and R3+R(i=N,N-diethylamine
This was similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3!4-dihydronapth-2-yl)-acrylic acid and N,N-diethylamine. Yield (86%), analysed for C17H20BrNO; calcd C 61.09 H 6.03 Br 23.91 N 4.19%, found C 61.97 H 6,14 Br 24.01 N 4.24%.
Example 32
Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid immidozolide of formula 1 where
Figure imgf000044_0002
R=CH2-Ph and R5+R6=immidazole.
This was' similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid and immidazole Yield (81%), analysed for Ci6H17BrN2O; calcd C 57.67 H 5.14 Br 23.98 N 8.41 %, found C 57.93 H 5.19 Br 24.04 N 8.49%.
Example 33
Synthesis of 3-(l-bromo-3,4-dihydronapth-2-yl)-acryIic acid /?-hydroxypiperizide of formula 1 where
Figure imgf000045_0001
This was similarly prepared, as described in example 24, except the starting materials were 3-(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid and /?-hydroxypiperidine. Yield (90%), analysed for Ci8H20BrNO2; calcd C 59.68 H 5.56 Br 22.06 N 3.87%, found C 61.81 H 5.61 Br 22.1 I N 3.93%.
Example 34
Synthesis of 3-(3,4-dihydronaphth-2-yl)-acrylic acid isobutylamide of formula 1 where Ri=R2=R3=Ri=H, R=H and R5+R6=isobutylamine. 1) Preparation of 1,2,3,4-tetrahydronaphth-l-oL
To a chilled solution of α-tetralone (7.6g, 52 mmol) in methanol (100 ml), sodium borohydride (500 mg) was added in 2 hour maintaining temperature 0-5 0C and after the completion of the reaction (monitored by TLC), the volume of the contents was reduced to one fifth, diluted with ice cold water (120 ml) and extracted with ether, the organic layer was washed with water and dried over anhydrous sodium sulfate, concentrated on a rotavapour under reduced pressure to give a colourless viscous oily compound 1,2,3,4- tetrahydronaphth-1-ol (7.1g, 93 %). Analysed for Ci0Hi2O; calcd C 81.01 H 8.16%, found C 82.11 H 8.21 %. 2) Preparation of 2-formyl-3,4-dihydronaphthalene.
To a chilled solution of 1,2,3,4-tetrahydronaρhth-l-ol (7g, 47.3 mmol) in dry DMF (25.0 ml), POCl3 (15.0 ml) was added drop wise at 0-5 0C for lhr and the resulting mixture was stirred at room temperature for 24 hrs. The reaction mixture was poured into ice-cold water (150 ml) and pH maintained by adding 0.5N NaOH solution and the reaction mixture extracted with ethyl acetate (3x100 ml). The combined organic layer washed with water (3x30 ml), dried over anhydrous sodium sulfate and evaporated on rotavapour to give crude product, which was purified on silica gel column using pet. ether 60-80 0C as an eluent to gave pale yellow compound 2-formyl-3,4-dihydronaphthalene (7.2 g, 96 %), analysed for CnHi0O; calcd C 83.53 H 6.37%, found C 84.44 H 6.41%.
3) Preparation of 3-(3,4-dihydronaphth-2-yI)-acrylic acid
5 To a mixture of 2-formyl-3,4-dihydronaphthalene (7.Og, 49.3 mmol) in pyridine (20 ml) and piperidine (3 nil), malonic acid (9.2 g 1.2 eq) was added and the reaction mixture kept at room, temperature for 52 hrs and then heated on water bath for 6 hrs. The contents were acidified with 50% HCl solution and the resulting precipitate filtered, washed with water and crystallized in pet. ether: ethyl acetate to yield pure product 3-(3,4-dihydronaphth-2-yl)- LO acrylic acid (9.1g. 93 %), analysed for Ci3H12O2; calcd C 77.98 H 6.04%, found C 78.76 H 6.11%. 1H NMR (200 MHz CDCl3): δ 2.92 (2H, t, J=Hz, CH2CH2), 2.52 (2H, t, J=Hz, CH2CH2), 6.00 (IH, d, J=Hz, CH=CHCO), 6.81 (IH3S, CH=C), 7.78 (4H, m, 4xAr-H), 8.21 (IH, d, J=Hz, CH=CHCO).
4) Preparation of 3-(3,4-dihydronapth-2-yI)-acryIic acid isobutylamide..
5 To the compound 3-(3,4-dihydronaphth-2-yl)-acrylic acid (200mg, 1 mmol) in dry benzene (20 ml) added freshly distilled thionyl chloride (1.0 ml) and refluxed for lhr, excess of thionyl chloride removed in vacuo and thereafter condensed with dry benzene solution of isobutyl amine (1.0 ml) and stirred for 30 min. The organic layer was washed with water (2x25 ml), dried over anhydrous sodium sulfate, concentrated to give crude product, which
!0 was purified on silica gel column using pet. ether: ethyl acetate (92:8) as an eluent to gave title compound (210mg, 82%) analysed for Ci7H20NO; calcd C 79.96 H 8.29 N 5.49%, found. C 80.84 H 8.33 N 5.55 %. 1H NMR (200 MHz CDCl3): δ 0.94 (6H, d, J=6.70 Hz, CH(CH3)2), 1.81 (1Η, m, CΗ(CΗ3)2), 2.43 (2H, t, J=8.0 Hz, CH2CH2), 2.88 (2H, t, J=8.0 Hz5 CH2CH2), 3.20 (2H, d, J=6.5 Hz, CH2CH), 5.96 (IH, d, J=15.32 Hz, CH=CHCO), 6.73
•5 (1Η, s, CH=C), 7.1 (4H, m, Ar-H)5 7.43 (IH5 d, J=I 5.33 Hz, CH=CHCO).
Example 35
Synthesis of 3-(3,4-dihydronaphth-2-yl)-acrylic acid N,N-diisopropylamide of formula
1 where
Figure imgf000046_0001
iO This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronapth-2-yl)-acrylic acid and N,N-diisopropylamine. Yield (94%), analysed for Ci9H25NO; calcd C 80.52 H 8.89 N 4.90%, found. C 81.43 H 8.93 N 4.97 %.
Example 36
Synthesis of 3-(3,4-dihydronaphth-2-yl)-acrylic acid piperidide of formula 1 where 5
Figure imgf000047_0001
and R5+R6=piperidide.
This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronapth-2-yl)-acrylic acid and piperidine. Yield (84%), analysed for C18H2INO; calcd C 80.86 H 7.92 N 5.24%, found C 81. "77 H 7.99 N 5.28 %. 1H NMR (200 MHz CDCl3): δ 1.63 (6H, bs, 3xCH2), 2.51 (2H, t, J=8.0 Hz, CH2CH2), 2.86 (2H, t, J=8.0 Hz, 0 CH2CH2), 3.55 (4H, m, 2xCH2), 671 (IH, d, J=15.32 Hz, CH=CHCO), 7.16 (4H, m, Ar-H), 7.45 (IH, d, J=15.33 Hz, CH=CHCO).
Example 37
Synthesis of 3-(3,4-dihydronaphth-2-yl)-acrylic acid pyrrolidide of formula 1 where 15 Ri=R2=R3=R4=H, R=H and R5+R6=pyrrolidine.
This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronapth-2-yl)-acrylic acid and pyrrolidine. Yield (94%), analysed for Cj7Hi9NO; calcd C 80.86 H 7.56 N 5.53%, found C 81.66 H 7.61 N 5.60%.
:0 Example 38
Synthesis of 3-(3,4-dihydronaphth-2-yl)-acrylic acid niorpholine of formula 1 where
Figure imgf000047_0002
and R5+R6=morpholine.
This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronapth-2-yl)-acrylic acid and morpholine. Yield (90%), analysed for 5 C17H19NO2;'calcd C 75.81 H 7.11 N 5.20%, found. C 76.77 H 7.18 N 5.27%.
Example 39
Synthesis of 3-(3,4-dihydronaphth-2-yl)-acrylic acid p-methoxyanilide of formula 1 where R1=R2=R3=RjI=H, R=H and R5+R6=p-methoxyaniIine.
0 This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronapth-2-yl)-acrylic acid and />-methoxyaniline. Yield (90%), analysed for C20Hi9NO2; calcd C 78.66 H=6.27 N 4.59%, found. C 79.44 H 6.31 N 4.62%.
Example 40
Synthesis of 3-(3, 4-dihydronaphth-2-yl)-acrylic acid w-octylamide of formula 1 where
Figure imgf000048_0001
This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronapth-2-yl)-acrylic acid and 7?-octylamine. Yield (90%), analysed for C21H29NO;,calcd C 80.98 H 9.38 N 4.50% found. C 81.99 H 9.43 N 4.58%.
Example 41
Synthesis of 3-(3,4-dihydronaphth-2-yl)-acrylic acid N,N-diethylamide of formula 1 where R1=R2=R3=R4 =H, R=H and Rs+R6=N,N-diethylamine.
This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronapth-2-yl)-acrylic acid and N.N-diethylamine. Yield (90%), analysed for CnH21NO; calcd C 79.96 H 8.29 N 5.49% found.C 80.77 H 8.33 N 5.54%.
Example 42
Synthesis of 3~(3,4-dihydronaphth-2-yl)-acrylic acid anilide of formula 1 where
Figure imgf000048_0002
R2=R3=Ri=H, R=H and Rs+R6=aniline. This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronapth-2-yl)-acrylic acid and aniline. Yield (90%), analysed for C19H17NO; calcd C 82.88 H 6.22 N 5.09%, found C 83.77 H 6.27 N 5.18%.
Example 43 Synthesis of 3-(3,4-dihydronaphth-2-yl)-acrylic acid 3,4-methelenedioxy anilide of formula 1 where
Figure imgf000048_0003
and R5+R<;=3,4-methyIenedioxy aniline.
This was similarly prepared, as described in example 34, except the starting materials were 3-(3,4-dihydronaρth-2-yl)-acrylic acid and 3,4-methylenedioxy aniline. Yield (90%), analysed for C20Hi7NO3; calcd C 75.22 H 5.37 N 4.39%, found C 76.11 H 5.42 N 4.44%. Example 44
Synthesis of 3-(l-benzyl-3,4-dihydronaphth-2-yl)-acrylic acid λi-octylamide of formula
1 where Ri=R2=R3=R4=H, R=CH2-Ph and R5+R6=«-octyIamine
1) Preparation of l-(benzyl)-tetrahydro-naphth-l-ol.
To an ethereal solution of Grignard reagent prepared from magnesium metal and benzyl bromide was added α-tetralone (15.Og, lOOmmol) in one hour and then the reaction mixture worked up by pouring . the contents intol% aqueous ammonium chloride solution, the organic layer separated and the aqueous layer extracted with solvent ether (4x100 ml), the combined Organic layer washed with water (3x20 ml), dried over anhydrous sodium sulphate, concentrated to give the title compound yield (17.5g, 70%),- analysed for Ci7HIsO; calcd C 85.67 H 7.61%, found C 85.95 H 7.69%.
2) Preparation of l-benzyl-2-formyI-3,4-dihydronaphthalene.
To a chilled solution of l-(benzyl)-tetrahydro-naphth-l-ol (1Og, 42 mmol) in dry DMF (35 ml), POCl3 (16.2 ml) was added drop wise at 0-5 0C for one hr and the resulting mixture was • stirred at room temperature for 45 hrs. The reaction mixture was poured into ice-cold water (250 ml) containing sodium acetate (15 g). The contents were extracted with ethyl acetate (3x100 ml). The combined extracts were pooled and washed with water (3x50 ml), dried over anhydrous sodium sulfate and concentrated on rotavapour to give crude product, which was purified on silica gel column using pet.ether 60-80 0C: ethyl acetate (90:10) as an eluent to gave yellow oily liquid compound l-benzyl-2-formyl-3, 4-dihydronaphthalene - (9.1g, 86%), analysed for Ci8H16O; calcd C 87.06 H 6.49%, found C 87.47 H 6.59%.
3) Preparation of 3-(l-benzyl-3, 4-dihydronaphth-2-yl)-acrylic acid.
To a mixture of l-benzyl-2-formyl-3, 4-dihydronaphthalene (8.5g, 32.7 mmol) in pyridine (25 ml) and piperidine (5.0 ml), malonic acid (I4.25g, 1.2 eq) was added and kept the reaction mixture at room temperature for 72 hrs. Later the reaction mixture was heated on water bath for 6 hrs, and poured the contents in to a conical flask. To this 50% aqueous HCl solution was added to precipitate the compound 3-(l-benzyl-3, 4-dihydronaρhth-2-yl)- acrylic acid. The precipitates were filtered and crystallized in pet. ether/ethyl acetate to yield pure 3-(l-benzyl-3, 4-dihydronaρhth-2-yl)-acrylic acid (7.5g, 78%), analysed for C2OHi8O2; calcd C 82.73 H 6.25%, found C 82.98 H 6.32%. 4) Preparation of 3-(l-benzyl-3,4-dihydronaphth-2-yl)-acrylic acid w-octyl amide.
To the compound 3-(l-benzyl-3, 4-dihydronaphth-2-yl)-acrylic acid (500mg, 2.01mmol). in dry benzene (20 ml) added freshly distilled thionyl chloride (1.0 ml) and refluxed for one hr, excess of thionyl chloride removed in vacuo and thereafter condensed with dry benzene solution of o.ctylamine (1.0 ml) and stirred for 30 min. The organic layer washed with water (2x25ml), dried over anhydrous sodium sulfate and evaporated on rotary evaporator to give crude product, which was purified on silica gel column using pet. ether 60-80 0C: ethyl acetate (85:15) as an eluent to gave title compound (620mg, 76%), analysed for C28H35NO; calcd C 83.74 H 8.78 N 3.49%, found C 84.02 H 8.84 N 3.56%. ' * ■
Example 45
Synthesis of 3-(l-benzyl-3,4-dihydronaphth-2-yl)-acrylic acid piperidide of formula 1 where Ri=R2=R3=Ri =H, R=CH2-Ph and R5+R6=piperidine. This was similarly prepared, as described in example 44, except the starting materials were 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid and piperidine. Yield (88%), analysed for C25H27NO; calcd C 83.99 H 7.61 N 3.92%, found C 84.61 H 7.70 N 4.40%:
Example 46
Synthesis of 3-(l-benzyl-3,4-dihydronaphth-2-yl)-acrylic acid pyrrolidine of formula 1 WhCrB R1=R2=R3=R4=H, R=CH2-Ph and R5+R6=pyrrolidine.
This was similarly prepared, as described in example 44, except the starting materials were 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid and pyrrolidine. Yield (88%), analysed for C24H25NO; calcd C 83.93 H 7.34 N 4.08%, found C 84.41 H 7.40 N 4.11%.
Example 47
Synthesis of 3-(l-benzyl-3,4-dihydronaphth-2-yl)-acrylic acid morpholide of formula 1 where R1=R2=R3=Rj=H, R=CH2-Ph and R5+R6=morpholine.
This was similarly prepared, as described in example 44, except the starting materials were 3-(l-benzyl-3,4-dihydiOnapth-2-yl)-acrylic acid and morpholine. Yield (79%), analysed for C24H25NO2; calcd C 80.19 H 7.01 N 3.90%, found C 80.91 H 7.08 N 3.98%. Example 48
Synthesis of 3-(l-benzyl-3,4-dihydronaphth-2-yl)-acrylic acid isobutylamide of formula
1 where
Figure imgf000051_0001
This was similarly prepared, as described in example 44, except the starting materials were 5 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid and isobutylamine. Yield (92%), analysed for C24H27NO; calcd C 83.44 H 7.88 N 4.05%, found C 83.91 H 7.92 N 4.11%.
Example 49
Synthesis of 3-(l-benzyI-3,4-dihydronaphth-2-yI)-acrylic N,N-diisopropyIamide of 10 formula 1 where
Figure imgf000051_0002
This was similarly prepared, as described in example 44, except the starting materials were 3-(i-benzyl-3,4-dihydronapth-2-yl)-acrylic acid and N,N-diisopropylamine. Yield (82%), analysed for C26H3iNO; calcd C 83.60 H 8.37 N 3.75%, found C 83.99 H 8.42 N 3.80%.
15 Example 50
Synthesis of 3-(l-benzyl-3,4-dihydronaphth-2-yl)~acrylicacid /j-hydroxy piperidine of formula 1 where
Figure imgf000051_0003
piperidine
This was similarly prepared, as described in example 44, except the starting materials were 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid and /?-hydroxypiperidine. Yield (72%), >0 analysed for C25H37NO2; calcd C 80.40 H 7.29 N 3.75%, found C 80.69 H 7.32 N 3.80%.
Example 51
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid piperidide of formula 1 where R1=R4=H, R2=R3=OCH3, R=H and Rg+R6==piperidine. '.5 1) Preparation of 4~allyl-i,2-dimethoxy benzene.
A solution comprising of methyl eugeonol (1Og, 60.97 mmol) and CH3I (9 ml) in dry acetone (50. ml) was refluxed at 60 C for 24 hrs. The reaction mixture volume was reduced to one fourth on rotavapour, contents cooled and diluted with water (150 ml), extracted with ethyl acetate (3x100 ml). The combined organic layer washed with water (3x30 ml), dried 0 over anhydrous sodium sulfate and evaporated on rotavapour to give crude product, which was purified on silica gel column using pet. ether 60-80 0C as an eluent to gave compound 4-allyl-l,2-dimethoxy benzene (9.3g, 85 %), analysed for CnHi4O2; calcd C 74.13 H 7.92%, found C 75.03 H 7.99%.
2) Preparation of 6,7-dimethoxy-2-formyl-3,4-dihydro-naphthalene
To a chilled solution of 4-allyl-l,2-dimethoxy benzene (9.Og, 50.96 mmol) in dry DMF (15 ml), POCl3 (13 ml) was added drop wise at 0-5 0C for 1 hr and the resulting mixture was stirred at room temperature for 72-90 hrs. The reaction mixture was poured into ice-cold water (150 ml) and pH 7.0 attained by adding IN NaOH solution' and the reaction mixture extracted with ethyl acetate (5x100 nil). The combined organic layer washed with water (3x30 ml), dried over anhydrous sodium sulfate and evaporated on rotavapour to give crude product, which was purified on silica gel column using pet. ether 60-80 0C: ethyl acetate (80:20)" as an eluent to gave compound 6,7-dimethoxy-2-formyl-3,4-dihydro-naphthalene (3.2g, 29%), analysed for C13Hi4O3; calcd C 71.54 H 6.67%, found C 72.44 H 6.71%. 1H NMR (CDCl3): δ 2.55 (2H, t, J=8.3 Hz), 2.82 (2H, t, J=8.3 Hz), 3.91 (6H, m), 6,74 (IH, s), 6.81 (IH, s), 7.20 (IH, s). 3) Preparation of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-acrylic acid.
To a stirring solution of ylide (11.5g) (prepared from triphenylphospine and ethyl bromoacetate) containing sodium hydride (1.Og) was added benzene solution of 6,7- dimethoxy-3,4-dihydro-naphthalene-2-carbaldehyde (5.5g, 25 mmol) and after 24 hours an additional amount of sodium hydride (0.5g) was added. The reaction mixture was stirred for 72 hours at 40 0C. On cooling, the contents were diluted with ethyl acetate (100 ml) to quench unused sodium hydride, then diluted with water (200 ml), organic layer separated and the aqueous layer extracted with ethyl acetate (2x100 ml). The combined organic layer washed with water (2x50 ml) and concentrated under reduced pressure. The crude 3-(6,7- dimethoxy-3,4-dihydro-naphtha-2-yl)-acrylate obtained above was hydrolysed without purification- in 10% methanolic potassium hydroxide solution and after usual work up ,the acid obtained was purified on silica gel column using pet. ether 60-80 0C: ethyl acetate (75:25) as an eluent to gave compound 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-acrylic acid (5.59g/86%), analysed for Ci5Hi6O4; calcd C 69.22 H 6.2%, found C 70.12 H 6.27%. 1H NMR (CDCl3): δ 2.49 (2H, t, J=8.20 Hz), 2.84 (2H, t, J=8.20 Hz), 3.86 (6H, m), 5.94 (IH, d, J=15.5 Hz); 6.71 (3H, m), 7.56 (IH, d, j=15.5 Hz). . 4) Preparation of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid piperidide
To the compound 3 -(6, 7-dimethoxy-3,4-dihydronaphth-2-yl)-acrylic acid (lOOmg, 0.384 mmol) in dry benzene (15 ml) added freshly distilled thionyl chloride (1.0 ml) and refluxed for 1 hr, excess of thionyl chloride evaporated in vacuo and thereafter condensed with dry benzene solution of piperidine (1.0 ml) and stirred for 30 min. The organic layer washed with water (2x25 ml), dried over anhydrous sodium sulfate, concentrated to give crude product, which was purified on silica gel column using pet. ether: ethyl acetate (85:15) as an eluent to gave title compound (1 lOmg, 88%) analysed for C2OH25NO3; calcd C 7337 H 7.70 N 4.28%, found C 74.21 H 7.78 N 4.31%. 1HNMR (CDCl3): δ 1.60 (6H, m), 2.47 (2H, t, J=8.15 Hz), 2.86 (2H, t, J=8.15 Hz), 3.59 (4H, m), 3.87 (6H, m), 6.39 (IH, d, J=15.1 Hz), 6.66 (3H3 m), 7.41 (IH, d, J=I 5.1 Hz).
Example 52
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yI)-propenoic acid morpholide of formula 1 where Ri=R4=H, R2=R3=OCH3, R=H, R3+R6=morpholine.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6, 7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and morpholine. Yield (80%), analysed for C19H23NO4; calcd C 69.28 H 7.04 N 4.25%, found C 70.11 H 7.10 N 4.31%. 1H NMR (CDCl3, δ): 2.47 (2H, t, J=8.2 Hz), 2.82 (2H, t, J=8.2 Hz), 3.71 (8H, m), 3.87 (6H, m), 6.32 (IH, d, J=15.1 Hz), 6.66 (2H, m), 7.11 (IH, d), 7.66 (IH, d, J=15.1 Hz).
Example 53
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid pyrrolidine of formula 1 where Ri=R4=H, R2=R3=OCH3, R=H and Rs+Rβ-pyrrolidine. This was also similarly prepared, as described in example 51, except the starting materials was 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and pyrrolidine. Yield (80%), analysed for Ci9H23NO3; calcd C 72.82 H 7.40 N 4.47%, found C 73.77 H 7.48 N 4.51%.
Example 54
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid /i-octylamide of formula 1 where R1=R4=H, R2=Ra=OCH3, R=H and R5+R<;=w-octylamine.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and n-octylamine. Yield (88%), •analysed for C23H33NO3; calcd C 74.36 H 8.95 N 3.77%, found C 74.97 H 9.02 N 3.81%. 1H NMR (CDCl3): δ 0.88 (3H5 1, J=63 Hz), 1.24 (1OH, m), 1.54 (2H, m), 2.42 (2H, t, J= 8.2 Hz), 2.82 (2H, t, J=8.2 Hz), 3.35 (2H, t, J=6.30 Hz), 5.90 (IH, d, J=15.2 Hz), 6.65 (2H, m), 7.09 (IH, d), 7.41 (IH, d, J=I 5.23 Hz).
Example 55 Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid isobutylamide of formula 1 where
Figure imgf000054_0001
isobutylamine.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and isobutylamine. Yield (90%), analysed for Ci9H25NO3; calcd C 72.35 H 7.99 N 4.44%, found C 72.99 H 8.08 N 4.49%. 1H NMR (CDCl3): δ 0.96 (6H, m), 1.83 (IH, m), 2.44 (2H, t, J=8.3 Hz), 2.77 (2H, t, J=8.3 Hz), 3.23 (2H, t, J=6.5 Hz), 5.90 (IH, d, J=I 5.3 Hz), 6.67 (2H, m), 7.12 (IH, d), 7.68 (IH, d, J=I 5.3 Hz).
Example 56 Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid /irbutylamide of formula 1 where Ri=R4=H, R2=R3=OCH3, R=H and R3+R6=n-butylamine.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-piOpenoic acid and π-butylamine. Yield (80%), analysed for Ci9H25NO3; calcd C 72.35 H 7.99 N 4.44%, found C 73.11 H 8.06 N 4.51%.
Example 57
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid λϊ-propylamide of formula 1 where Ri=R4=H, R2=R3=OCH3, R=H and R5+R6=«-propylamine.
This w.as similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yI)-piOpenoic acid and ^-propylamine. Yield (80%), analysed for Ci8H23NO3; calcd C 71.73 H 7.69 N 4.65%, found C 72.44 H 7.75 N 4.70%. Example 58
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid isopropylamide of formula 1 where Ri=R4=H, R2=Rs=OCH3, R=H and R5+R6= iopropylamine.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and isopropyl amine. Yield (80%), analysed for C18H23NO3; calcd C 71.73 H 7.69 N 4.65%, found C 72.45 H 7.74 N 4.71%.
Example 59 Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid N,N- diisopropylamide of formula 1 where Ri=R4=H, R2=R3=OCH3, R=H and R5H-Re= N,N- diisopropylamine.
This was similarly prepared, as described in example 51, except the starting materials are 3- (6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and N,N-diisopropylamine. Yield (85%), analysed for C21H29NO3; calcd C 73.44 H 8.51 N 4.08%, found C 74.11 H 8.59 N 4.14%.
Example 60
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid anilide of formula 1 where Ri=R4=H, R2=R3=OCH3, R=H and R5+R<;=aniline.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and aniline. Yield (85%), analysed for C2iH2iNO3; calcd C 75.20 H 6.31 N 4.18%, found C 76.11 H 6.39 N 4.24%.
Example 61
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid o-methyl anilide of formula 1 where Ri=R4=H, R2=R3=OCH3, R=H and R5+R6=ø-methyl aniline.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and ø-methyl aniline. Yield (89%),' analysed for C22H23NO3; calcd C 76.62 H 6.63 N 4.01%, found C 77.23 H 6.71 N 4.09%. Example 62
Synthesis of 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid p-methyl anilide of formula 1 where
Figure imgf000056_0001
R=H and
Figure imgf000056_0002
aniline.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and p-methyl aniline. Yield (72%), analysed for C22H23NO3; calcd C 76.62 H 6.63 N 4.01%, found C 77.45 H 6.69 N 4.11%.
Example 63 Synthesis of 3~(6,7-dimethoxy-3,4-dihydronaphth~2-yl)- propenoic acid /?-methoxy anilide of formula 1 where
Figure imgf000056_0004
R=H and
Figure imgf000056_0003
aniline.
This was similarly prepared, as described in example 51, except the starting materials are 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid and j>methoxy aniline. Yield (82%), analysed for C22H23NO4; calcd C 72.31 H 6.34 N 3.83%, found C 72.98 H 6.40 N 3.89 %.
Example 64
Synthesis of 5-(l-chloro-3,4-dihydronaphth-2-yl)-4-methyl-2E,4JE'-pentadienoic acid- isobutylamide of formula 2 where Ri=R2=R3=R4=H, R=Cl, R5+R<;=isobutenylamine and R7=methyl.
1) Preparation of l-chloro-2-formyl-3,4-dihydronaphthalene
To a chilled solution of α-tetralone (25g, 170 mmol) in dry DMF (15 ml), POCl3 (15.75 ml) was added drop wise at 0-5 0C for one lir. and the resulting mixture was stirred at room temperature for 24 hrs. The reaction mixture was poured into ice-cold water (150 ml) containing sodium acetate (25 g). The reaction mixture was extracted with ethyl acetate (3x200 ml). The combined extracts were washed with water (3x50 ml), dried over anhydrous sodium sulfate and evaporated on rota vapour to give crude product, which was purified on silica gel column using pet. ether 60-80 0C as an eluent to gave yellow viscous oil compound l-chloro-2-formyl-3,4-dihydronaphthalene (31 g, 95%), analysed for C, ,H9ClO; calcd C 68.58 H 4.71 Cl 18.40%, found C 69.45 H 4.78 Cl 18.46%. 2) Preparation of l-(l-chloro-3,4-dihydronaphth-2-yl)-propan-l-ol.
To an ethereal solution of. Grignard reagent prepared from magnesium metal and ethyl iodide was added compound l-chloro-2-formyl-3,4-dihydronaphthalene (1Og, 52mmol) in one hour and then the reaction mixture worked up by pouring the contents into 1% aqueous ammonium chloride solution, the organic layer separated and the aqueous layer extracted with solvent ether (4x100 ml), the combined organic layer washed with water (3x20 ml), dried over anhydrous sodium sulphate, concentrated to give the. product l-(l-chloro-3,4- dihydronaρhth-2-yl)-propan-l-ol as an viscous oil (7.6g, 66%), analysed for CnHi5ClO; calcd C 70.11 H 6.79 Cl 15.92%, found C 71.55 H 6.84 Cl 15.99%. 3) Preparation of 3-(l-chloro-3,4-dihydronaphth-2-yl)-2-methyl-propenal.
To a chilled solution of l-(l-chloro-3,4-dihydronaphth-2-yl)-propan-l-ol (5g, 22.5 mmol) in dry DMF (25 ml), POCl3 (11 ml) was added drop wise at 0-5 0C for lhr and the resulting mixture was stirred at room temperature for 24 hrs. The reaction mixture was poured into ice-cold water (250 ml) containing sodium acetate (15 g). The contents were extracted with ethyl acetate (3x100 ml). The combined extracts were pooled and washed with water (3x50 ml), dried over anhydrous sodium sulfate and concentrated on rota vapour to give crude product, which was purified on silica gel column using pet. ether 60-80 0C: ethyl acetate (98:2) as an eluent to gave yellow oily liquid compound 3-(l-chloro-3,4-dmydroriaphth-2- yl)-2-methyl-propenal (4.5g, 86%), analysed for C14H13ClO; calcd C 72.26 H 5.63 Cl 15.24%, found C 72.97 H 5.69 Cl 15.31%.
4) Preparation of 5-(l-chloro-3,4-dihydronaphth-2-yl)-4-methyl-2£',4£'-pentadienoic acid.
To a stirring solution of ylide (8.5g) (prepared from triphenylpho spine and ethyl bromoacetate) containing sodium hydride (1.Og) was added benzene solution of 3-(l-chloro- 3,4-dihydronaphth-2-yl)-2-methyl-propenal (3.5 g,15 mmol) and after 24 hours an additional amount of sodium hydride (0.5g) was added. The reaction mixture was stirred for 72 hours at 40 0C. On cooling the contents were diluted with ethyl acetate (100 ml) to quench unused sodium hydride, and then diluted with water (200 ml), organic layer separated and the aqueous layer extracted with ethyl acetate (2x100 ml). The combined organic layer washed with water (2x50 ml) and concentrated under reduced pressure. The crude 5-(l-chloro-3,4- dihydronapl)th-2-yl)-4-methyl-2E,4E-ρentadienoic acid ester thus obtained was hydrolysed without purification in 10% methanolic potassium hydroxide solution and after usual work up, the acid obtained was purified on silica gel column using pet. ether 60-80 C: ethyl acetate (75:25) to give 5-(l-chloro-3,4-dihydronaphth-2-yl)-4-methyl-2£'s4£'-pentadienoic acid (3.2g, 80%), analysed for Ci6Hi5ClO2; calcd C 69.95 H 5.50 Cl 12.90%, found C 70.77 H 5.59 Cl 12.98%.
5) Preparation of 5-(l-chloro-3,4-dihydronaphth-2-yl)-4-methyI-2E,4E-pentadienoic acid isobutylamide.
To the compound 5-(l-chloro-3,4-dihydronaphth-2-yl)-4-methyl-2E,4E-pentadienoic acid (500mg, 1.82 mmol) in dry benzene (40 ml) added freshly distilled thionyl chloride (1.5 ml) and refluxed for lhr, excess of thionyl chloride removed in vacuo and thereafter condensed with dry benzene solution of isobutyl amine (1.0 ml) and stirred for 30 min. The organic layer washed with water (2x25 ml), dried over anhydrous sodium sulfate, concentrated to give crude product, which was purified on silica gel column using pet. ether: ethyl acetate (92; 8) as an eluent to give title compound (500mg, 83%), analysed for Ca0H24ClNO; calcd C 72.82 H 7.33 Cl 10.75 N 4.25, found C 73.33 H 7.39 Cl 10.81 N 4.30%.
Example 65
Synthesis of 5-(l-chloro-3,4-dihydronaphth-2-yI)-4-methyI-2E,4E-pentadienoic acid N,N-diisopropylamide of formula Ib where Ri=R2=Ra=Ri=H, R=Cl, R5+Rβ=N,N- diisopropylamine and R7=methyl.
The title compound was similarly prepared, as described in example 64, except the starting materials are 5-(l-chloro-3,4-dihydronaphth-2-yl)-4-methyl-2E,4E-pentadienoic acid and N,N-diisppropylamine. Yield (90%), analysed for C22H28ClNO; calcd C 73.83 H 7.89 Cl 9.91 N 3.91%, found C 74.17 H 7.92 Cl 9.99 N 3.98%.
Example 66
Synthesis of 5-(l-chloro-3,4-dihydronaphth-2-yl)-4-methyl-2JE',4£'-pentadienoic acid piperidide of formula Ib where
Figure imgf000058_0001
and
R7=methyl. The title compound was similarly prepared, as described in example 64, except the starting materials are 5-(l-chloro-3,4-dihydro-naρhth-2-yl)-4-methyl-penta-2,4-dienoic acid and piperidine. Yield (82%), analysed for C2iH24ClNO; calcd C 73.78 H 7.08 Cl 10.37 N 4.10%, found C 74.11 H 7.11 Cl 3.41 N 4.17%.
Example 67
Synthesis of 5-(l-chloro-3,4-dihydronaphth-2-yI)-4-methyl-22?,42T-pentadienoic acid pyrrolidide of formula Ib where
Figure imgf000059_0001
and
R7-methyl.
The title compound was similarly prepared, as described in example 64, except the starting materials are 5-(l-chloro-3,4-dihydronaphra-2-yl)-4-methyl-penta-22s,4i?- dienoic acid and pyrrolidine. Yield (94%), analysed for C20H22ClNO; calcd C 73.27 H 6.76 Cl 10.81 N 4.27%, found C 74.12 H 6.71 Cl 10.78 N 4.21%.
Example 68
Synthesis of 5-(l-chloro-3,4-dihydronaphth-2-yI)-4-methyl-2E,4JE'-pentadienoic acid morpholide of formula Ib where
Figure imgf000059_0002
and
R7=methyl.
The title compound was similarly prepared, as described in example 64, except the starting materials were 5-(l-chloro-3,4-dihydro-naphth-2-yl)-4-methyi-penta-2,4-dienoic acid and morpholine. Yield (90%), analysed for C20H22ClNO2; calcd C 69.86 H 6.45 Cl 10.31 N 4.07%, found C 70.12 H 6.49 Cl 10.39 N 4.13%.
Example 69a
Decrease in the MICs of Amikacin against Staphylococcus aureus, MRSA and other Gram positive bacteria when used in combination with compound of formula Ia where Ri=R2=R3=Ri=H, R=Cl and Rs+R<;=N,N-diisopropyIamine.
Minimum Inhibitory Concentration (MIC) of Amikacin alone and in combination with the above mentioned potentiator was performed against Staphylococcus aureus species, using method described in the study design. Sixteen to four fold reductions in MIC of Amikacin was observed in combination with the potentiator (Table Ia). Example 69b
Decrease in the MICs of Amikacin against Staphylococcus aureus, MRSA and other Gram positive bacteria when used in combination with compound of formula Ia where
Figure imgf000060_0001
R=CI and R5+R<i=N,N-diethyIamine. Minimum Inhibitory Concentration (MIC) of Amikacin alone and in combination with the above mentioned potentiator was performed against Staphylococcus aureus species, using method described in the study design. Sixteen to four fold reductions in MIC of Amikacin was observed in combination with the potentiator (Table Ib).
Example 69c
Decrease in the MICs of Amikacin against Staphylococcus aureus, MRSA and other Gram positive bacteria when used in combination with compound of formula Ia where
Figure imgf000060_0002
4-methylenedioxy-aniline.
Minimum Inhibitory Concentration (MIC) of Amikacin alone and in combination with the above mentioned potentiator was performed against Staphylococcus aureus species, using method described in the study design. Four to two fold reductions in MIC of Amikacin was observed in combination with the potentiator (Table Ic).
Example 69d Decrease in the MICs of Amikacin against Staphylococcus aureus, MRSA and . other Gram positive bacteria when used in combination with compound of formula Ia where
Figure imgf000060_0003
Minimum Inhibitory Concentration (MIC) of Amikacin alone and in combination with the above mentioned potentiator was performed against Staphylococcus aureus species, using method described in the study design. Sixteen to four fold reductions in MIC of Amikacin was observed in combination with the potentiator (Table Id).
Example 69e
Decrease in the MICs of Amikacin against Staphylococcus aureus, MRSA and other Gram positive bacteria when used in combination with compound of formula Ia where
Figure imgf000060_0004
Minimum Inhibitory Concentration (MIC) of Amikacin alone and in combination with the above mentioned potentiator was performed against Staphylococcus aureus species, using method described in the study design. A two to four fold reductions in MIC of Amikacin was observed in combination with the potentiator (Table Ie).
Example 69f
Decrease in the MICs of Amikacin against Staphylococcus aureus, MRSA and other Gram positive bacteria when used in combination with compound of formula laa [3- (l-chloro-l^jS^-tetrahydronapth-ϊ-yO-propanoic acid NjN-diisopropylamide]. Minimum Inhibitory Concentration (MIC) of Amikacin alone and in combination with the above mentioned potentiator was performed against Staphylococcus aureus species, using method described in the study design. A two fold reductions in MIC of Amikacin was observed in combination with the potentiator (Table If).
Example 69g
Decrease in the MICs of Amikacin against Staphylococcus aureus, MRSA and other Gram positive bacteria when used in combination with compound of formula laa [5- (l-chloro~ϊ,2,3,4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid piperidide].
Minimum Inhibitory Concentration (MIC) of Amikacin alone and in combination with the above mentioned potentiator was performed against Staphylococcus aureus species, .using method described in the study design.' A two fold reductions in MIC of Amikacin was observed in combination with the potentiator (Table Ig).
Example 70a Decrease in the MICs of Ciprofloxacin against Staphylococcus aureus, MRSA and other Gram positive bacteria used in combination with compound of formula Ia where
Figure imgf000061_0001
and R5+R6=N,N-diisopropylamine.
Minimum Inhibitory Concentration (MIC) of ciprofloxacin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Eight to two fold reductions in MIC of ciprofloxacin was observed in combination with the potentiator (Table 2a). Example 70b
Decrease in the MICs of Ciprofloxacin against Staphylococcus aureus, MRSA and other Gram positive bacteria used in combination with compound of formula Ia where
Figure imgf000062_0001
R=CI and R5+R<;=N,N-diethylamine. Minimum Inhibitory Concentration (MIC) of ciprofloxacin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Four to two fold reductions in MIC of ciprofloxacin were observed in combination with the potentiator (Table 2b).
Example 70c
Decrease in the MICs of Ciprofloxacin against Staphylococcus aureus, MRSA and other Gram positive bacteria used in combination with compound of formula Ia where Ri=R2=R3=R4=H, R=Cl and R5+R6=3,4-methylenedioxyaniline>
Minimum Inhibitory Concentration (MIC) of ciprofloxacin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Two fold reductions in MIC of ciprofloxacin were observed in combination with the potentiator (Table 2c).
Example 7Od Decrease in the MICs of Ciprofloxacin against Staphylococcus aureus, MRSA and other Gram positive bacteria used in combination with compound of formula Ia where Ri=R2=R3=Ri=H, R=CI, R5+R6=piperidine and R7=methyl.
Minimum Inhibitory Concentration (MIC) of ciprofloxacin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Four to two fold reductions in MIC of ciprofloxacin were observed in combination with the potentiator (Table 2d).
Example 7Oe
Decrease in the MICs of Ciprofloxacin against Staphylococcus aureus, MRSA and other Gram positive bacteria used in combination with compound of formula Ia where Ri=R2 =R3=R4=H, R=Cl and R5+R<;=isobutylamine. Minimum Inhibitory Concentration (MIC) of ciprofloxacin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Eight to two fold reductions in MIC of ciprofloxacin was observed in combination with the potentiator (Table 2e).
5
Example 7Of
Decrease in the MICs of Ciprofloxacin against Staphylococcus aureus, MRSA and other Gram positive bacteria used in combination with compound of formula ϊa where
Figure imgf000063_0001
[0 Minimum Inhibitory Concentration (MIC) of ciprofloxacin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Eight to two fold reductions in MIC of ciprofloxacin was observed in combination with the potentiator (Table 2f).
5 Example 7Og
Decrease in the MICs of Ciprofloxacin against Staphylococcus aureus, MRSA arid other Gram positive bacteria used in combination with compound of formula Ia where
Figure imgf000063_0002
Minimum Inhibitory Concentration (MIC) of ciprofloxacin alone and in combination with !0 the above mentioned potentiator was performed against bacterial species, using method described in the study design. Eight to two fold reductions in MIC of ciprofloxacin was observed in combination with the potentiator (Table 2g).
Example 7Oh
5 Decrease in the MICs of Ciprofloxacin against Staphylococcus aureus, MRSA and other Gram positive bacteria used in combination with compound of formula Ia where
Figure imgf000063_0003
R=H and R5+R6=N,N-diisoproρylamine.
Minimum Inhibitory Concentration (MIC) of ciprofloxacin alone and in combination with ■ the above mentioned potentiator was performed against bacterial species, using method 0. described in the study design. Two fold reductions in MIC of ciprofloxacin were observed in combination with the potentiator (Table 2h). Example 71a
Decrease in the MICs of rifampicin against M.tiiberculosis, M. avium and M. intracellure when used in combination with compound of formula Ia where
Figure imgf000064_0001
Minimum Inhibitory Concentration (MIC) of rifampicin alone and in combination with the above mentioned potentiator was performed against Mycobacterial species, using method described in the study design. Four to two fold reductions in MIC of rifampicin was observed in combination with the potentiator (Table 3a).
Example 71b
Decrease in the MICs of rifampicin against M.tuberculosis, M. avium and M. intracellure when used in combination with compound of formula Ia where
Figure imgf000064_0002
and R5+R6=N,N-diethyIamine.
Minimum Inhibitory Concentration (MIC) of rifampicin alone and in combination with the above mentioned potentiator was performed against Mycobacterial species, using method described in the study design. Four to two fold reductions in MIC of rifampicin was observed in combination with the potentiator (Table 3 b).
Example 71c Decrease in the MICs of rifampicin against M.tuberculosis, M. avium and M. intracellure when used in combination with compound of formula Ia where
Figure imgf000064_0003
R=Br and R5+R<;=isobutylamine.
Minimum Inhibitory Concentration (MIC) of rifampicin alone and in combination with the above mentioned potentiator was performed against Mycobacterial species> using method described in the study design. Eight to four fold reductions in MIC of rifampicin was observed in combination with the potentiator (Table 3c).
Example 71 d
Decrease in the MICs of rifampicin against M.tuberculosis, M. avium and M. intracellure when used in combination with compound of formula Ia where R1=R2=R3=Ri=H, R=H and R5+R6=N,N-diisopropylamine. Minimum Inhibitory Concentration (MIC) of rifampicin alone and in combination with the above mentioned potentiator was performed against Mycobacterial species, using method described in the study design. Four to two fold reductions in MIC of rifampicin was observed in combination with the potentiator (Table 3d). • ■ ' ' '
Example 72a
Decrease in the MICs of mupirocin against Staphylococcus aureus, and MRSA used in combination with compound of formula Ia where Rx=R2=Rs=Ri=H, R=Cl and R5+R6=N,N-diisopropylamine. Minimum Inhibitory Concentration (MIC) of mupirocin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Eight to two fold reductions in MIC of mupirocin was observed in combination with the potentiator (Table 4a).
Example 72b
Decrease in the MICs of mupirocin against Staphylococcus aureus, and MRSA isolates used in combination with compound of formula Ia where R1=R2=Rs-Ri -H, R=Cl and Rs+R6=N,N-diethyIamine.
Minimum Inhibitory Concentration (MIC) of mupirocin. alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Eight to foμr fold reductions in MIC of mupirocin was observed in combination with the potentiator (Table 4b).
Example 72c Decrease in the MICs of mupirocin against Staphylococcus aureus, and MRSA used in combination with compound of formula Ia where
Figure imgf000065_0001
and Rs+R6=3,4-methylenedioxy-aniline.
Minimum Inhibitory Concentration (MIC) of mupirocin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. A two fold reduction in MIC of mupirocin was observed in combination with the potentiator (Table 4c). Example 72d
Decrease in the MICs of mupirocin against Staphylococcus aureus, and MRSA isolates used in combination with compound of formula Ib where
Figure imgf000066_0001
R=CI,
Figure imgf000066_0002
5 Minimum Inhibitory Concentration (MIC) of mupirocin alone and in combination with the above mentioned potentiator was performed against bacterial species, using method described in the study design. Four to two fold reductions in MIC of mupirocin was observed in combination with the potentiator (Table 4d).
10 Example 72e
Decrease in the MICs of mupirocin against Staphylococcus aureus, and MRSA isolates used in combination with compound of formula Ia where Ri=R2=Rs=Ri=H, R=Br and
Minimum Inhibitory Concentration (MIC) of mupirocin alone and in combination with the 15 above mentioned potentiator was performed against bacterial species, using method described in the study design. Eight to two fold reductions in MIC of mupirocin was observed in combination with, the potentiator (Table 4e).
Example 72f
'.0 Decrease in the MICs of mupirocin against Staphylococcus aureus, and MRSA used in combination with compound of formula Ia where
Figure imgf000066_0004
and Rs+R<;=N,N-diisopropyϊamine.
Minimum Inhibitory Concentration (MIC) of mupirocin alone and in combination with the above mentioned potentiator was performed against bacterial species, Using method ,5 described in the study design. A two fold reduction in MIC of mupirocin was observed in combination with the potentiator (Table 4f).
Example 73a
Reduction in the dose requirement of Ciprofloxacin when used in combination with 0 compound of formula Ia where
Figure imgf000066_0005
diisopropylamine in systemic infection model of mice. The study was conducted to see the in vivo response of ciprofloxacin in combination with the above mentioned potentiator. The Swiss albino mice were infected intravenously with Staphylococcus aureus ATCC 29213 (107 CFU/mouse). The infected mice were divided in groups and each group consisted of 6 mice. The treatment consisted of one dose immediately after the infection followed by the next dose after a gap of 6 hrs. The result was recorded as number of survivals each day. The mice were observed for seven days and ED50 was determined after seven days of observation. The ED50 for ciprofloxacin was 9.2 mg/kg and in combination with the potentiator the ED50 for ciprofloxacin was reduced to 5.86 mg/kg.
Example 73b
Reduction in the dose requirement of Ciprofloxacin when used in combination with compound of formula Ia where
Figure imgf000067_0001
and R5+R<;=isobutylamine in systemic infection model of mice. The study was conducted to see the in vivo response of ciprofloxacin in combination with the above mentioned potentiator. The Swiss albino mice were infected intravenously with Staphylococcus aureus ATCC 29213 (107 CFU/mouse). The infected mice were divided in groups and each group consisted of 6 mice. The treatment consisted of one dose immediately after the infection followed by the next dose after a gap of 6 hrs. The result was recorded as number of survivals each day. The mice were observed for seven days and ED50 was determined after seven days of observation. The ED50 for ciprofloxacin was 9.2 mg/kg and in combination with the potentiator the ED50 for ciprofloxacin was reduced to 4.96 mg/kg.
Example 73c
Reduction in the dose requirement of Ciprofloxacin when used in combination with compound of formula Ia where
Figure imgf000067_0002
diisopropylamine in systemic infection model of mice.
The study was conducted to see the in vivo response of ciprofloxacin in combination with the above mentioned potentiator. The Swiss albino mice were infected intravenously with
Staphylococcus aureus ATCC 29213 (107 CFU/mouse). The infected mice were divided in groups and each group consisted of 6 mice. The treatment consisted of one dose immediately after the infection followed by the next dose after a gap of 6 hrs. The result was recorded as number of survivals each day. The mice were observed for seven days and ED50 was determined after seven days of observation. The ED50 for ciprofloxacin was 9.2
Y
5 mg/kg and in combination with the potentiator the ED50 for ciprofloxacin was reduced to 7.86 mg/kg.
Example 74
Reduction in the dose requirement of Mupirocin when used in combination with 10 compound of formula Ia where
Figure imgf000068_0001
diisopropylamine.
The study was conducted to see the in vivo response of Mupirocin in combination with the above mentioned potentiator. Wound area of 1.5 cm X 1.5 cm was created on the skin of mice by abrasion with sand paper. An inoculum of 108 CFU/ml of MRSA was applied on to 15 this wound area. The wound was treated for five days (two applications a day) with mupirocin cream and the other group with a formulation of mupirocin at l/4n concentration and compound of formula Ia. The combination of mupirocin with compound of formula Ia showed better wound healing (Table 5).
>0 Example 75
Increased accumulation and decreased efflux of ethidium bromide by compound of formula Ia where Ri=R2=R3=Rt=H, R=Cl and R5+R6=N,N-diisopropylamine in Staphylococcus aureus (wild type) and Ciprofloxacin mutant (CipR)
Measurement of the level of ethidium bromide accumulation and efflux in S. aureus ATCC
'.5 29213 (wild strain) and strain CIPr-l (ciprofloxacin-selected mutant) was based on a previously described method (Brenwald et.al, Antimicrobial Agents and Chemotherapy
1998; 42(8): 2032-2035). Briefly, for measurement of the level of accumulation, both the bacterial strains were grown overnight on Trypticase Soya Agar. Bacterial suspensions were prepared at an optical density at 550 nm of 0.2 in uptake buffer (NaCl, 110 mM; KCl, 7
0 mM; NH4C1, 50 mM; Na2HPO4, 0.4 mM; Tris base, 52 mM; glucose, 0.2% adjusted to pH
7.5 with HCl) and were then exposed to ethidium bromide at a concentration of 2μg/ml. The increase in fluorescence as ethidium bromide entered the cells was recorded fluorometrically with a Perkin-Elmer model LS50 spectrofluorimeter (excitation λ, 530 nm; emission, 600 nm) at 300C. The effect of compound of formula Ia where 3-(l-chloro-3,4-dihydronapth-2- yl)-acrylic acid N,N-diisopropylamide on the level of accumulation was determined in a similar way, except that compound of formula Ia was added to the uptake buffer at a concentration of 25 μg/ml (Figure IB).
For determining ethidium bromide loss, bacterial suspensions were prepared as described above and were exposed to ethidium bromide (2 μg/ml) in the presence of compound of formula 1 (25μg/ml) for 30 min at 37 0C. The cells were then pelleted by centrifugation and were re-suspended in fresh uptake buffer. The loss of ethidium bromide from the cells was measured as a decrease in fluorescence (Figure IB).
Table Ia. MICs of Amikacin alone and in combination with compound of formula Ia where
Figure imgf000069_0001
and R5+R6=N,N-diisopropylamine. (Figures in bold face show reduction in MIC).
Figure imgf000069_0002
Table Ib. MICs of Amikacin alone and in combination with compound of formula Ia where Ri=R2=R3=Ri=H, R=Cl and R5+R<;=N,N-diethylamine. (Figures in bold face show reduction in MIC)
Figure imgf000069_0003
Table Ic. MICs of Amikacin alone and in combination with compound of formula Ia where
Figure imgf000070_0001
(Figures in bold face show reduction in MIC)
Figure imgf000070_0003
Table Id. MICs of Amikacin alone and in combination with compound of formula Ia where Ri=R2=Rs=Ri=H, R=Br and Rs+R^isobutylamine. (Figures in bold face show reduction in MIC)
Figure imgf000070_0004
Table Ie. MICs of Amikacin alone and in combination with compound of formula Ia where
Figure imgf000070_0002
(Figures in bold face show reduction in MIC)
Figure imgf000070_0005
Table If. MICs of Amikacin alone and in combination with compound of 3-(l-chloro- l,2,3,4-tetrahydronapth-2-yl)-propanoic acid N,N-diisopropylamide. (Figures in bold face show reduction in MIC
Figure imgf000071_0002
Table Ig. MICs of Amikacin alone and in combination with compound of 5-(l-chloro- l,2,3,4-tetrohydronaphth-2-yl)-4-methyl-pentanoic acid piperidide. (Figures in bold face show reduction in MIC)
Figure imgf000071_0003
Table 2a. MICs of Ciprofloxacin alone and in combination with compound of formula [0 Ia where
Figure imgf000071_0001
(Figures in bold face show reduction in MIC)
Figure imgf000071_0004
Table 2b. MICs of Ciprofloxacin alone and in combination with compound of formula Ia where
Figure imgf000072_0001
=H, R=Cl and R5+R6=N,N-diethylamine. (Figures in bold face show reduction in MIC)
Figure imgf000072_0002
Table 2c. MICs of Ciprofloxacin alone and in combination with compound of formula Ia where Ri=R2=R3=R4=H, R=Cl and R5+R6=3,4-methylenedioxy-aniline. (Figures in bold face show reduction in MIC)
Figure imgf000072_0003
Table 2d. MICs of Ciprofloxacin alone and in combination with compound of formula Ib where Ri=R2=R3=R4=H, R=Cl, R5-HR<;=piperidine and R7=methyl. (Figures in bold face show reduction in MIC)
Figure imgf000072_0004
Table 2e. MICs of Ciprofloxacin alone and in combination with compound of formula Ia where R1=R2 =R3=R4 =H, R=Cl and R5+R6=isobutylamine. (Figures in bold face show reduction in MIC)
Figure imgf000073_0004
Table 2f. MICs of Ciprofloxacin alone and in combination with compound of formula Ia where
Figure imgf000073_0001
and Rs+R6=isobutylamine. (Figures in bold face show reduction in MIC)
Figure imgf000073_0005
Table 2g. MICs of Ciprofloxacin alone and in combination with compound of formula Ia where
Figure imgf000073_0002
R=Br and
Figure imgf000073_0003
(Figures in bold face show reduction in MIC)
Figure imgf000073_0006
Table 2h. MICs of Ciprofloxacin alone and in combination with compound of formula Ia Λvhere
Figure imgf000074_0001
R=H and R5+R6=N,N-diisopropylamine. (Figures in bold face show reduction in MIC)
Figure imgf000074_0003
Table 3a MIC of Rifampicin alone and in combination with compound of formula Ia where
Figure imgf000074_0002
R=CI and R5+R<;=N,N-diisopropylamine. (Figures in bold face show reduction in MIC)
[0
Figure imgf000074_0004
Table 3b MIC of Rifampicin alone and in combination with compound of formula Ia where Ri=R2=R3=R4 =H, R=Cl and R5+R6=N,N-diethyIamine. (Figures in bold face show reduction in MIC)
Figure imgf000074_0005
Table 3c MIC of Rifampicin alone and in combination with compound of formula Ia where
Figure imgf000075_0001
R=Br and R3+R6=isobutylamine. (Figures in bold face show reduction in MIC)
Figure imgf000075_0004
Table 3d MIC of Rifampicin alone and in combination with compound of formula Ia where
Figure imgf000075_0002
R=H and R5+R6=N,N-diisopropylamine. (Figures in bold face show reduction in MIC)
Figure imgf000075_0005
Table 4a. MICs of Mupirocin alone and in combination With compound of formula Ia where
Figure imgf000075_0003
(Figures in bold face show reduction in MIC)
Figure imgf000075_0006
Table 4b. MICs of Mupirocin alone and in combination with compound of formula Ia where Ri=R2=R3=RpH, R=CI and R5+R6=N,N-diethyIamine. (Figures in bold face show reduction in MIC)
Figure imgf000076_0002
Table 4c. MICs of Mupirocin alone and in combination with compound of formula Ia where Rt=R2=Ra=^=H, R=Cl and Rs+R6=3,4-methyIenedioxyanilinei (Figures in bold face show reduction in MIC)
10
Figure imgf000076_0003
Table 4d. MICs of Mupirocin alone and in combination with compound of formula Ib where
Figure imgf000076_0001
and R7=methyl. (Figures in bold face show reduction in MIC)
15
Figure imgf000076_0004
>0 Table 4e. MICs of Mupirocin alone and in combination with compound of formula Ia where Ri=R2=Ra=Ri=H, R=Br and R5+R6=isobutylamine. (Figures in bold face show reduction in MIC)
Figure imgf000077_0002
Table- 4f. JMICs of Mupirocin alone and in combination with compound of formula Ia where
Figure imgf000077_0001
(Figures in bold face show reduction in MIC)
10
Figure imgf000077_0003
[5
Advantages of the invention
The present invention discloses the preparation of a novel class of compounds Le 3-( unsubstitύted-or-substituted-3,4-dihydronaph-2yl-)-acrylic acid amides,5— (unsubstituted-or substituted-3,4-dihydronaph-2yl-)4-substitued-2E,4E-pentadionic acid amides and the saturated derivatives of these compounds ,the whole class of compounds not reported hitherto in the literature and their use as potentiators of antibiotics (i.e as efflux pump inhibitors of bacteria) when used in combination with antibiotic and tested against bacteria. The synthesised molecules have shown high potency in terms of lowering of MIC of the drug ciprofloxacin (thirtytwo fold) compared to the MIC levels of drug when used alone. These classes of compounds have shown potentiation of the in-vivo efficacy of the drugs when tested in in-vivo animal experimental models.
This is exemplified by a typical topical formulation of mupirocin in combination with the compound of formula 1 (where R1=R2=R3=R4=H, R=Cl and R5+R6=N,N-diisopropylamine) when used in defined ratio showed better efficacy than the original mupirocin in the mice dermal infection model (Table 5)

Claims

We Claim
1. A Compound of general formula I, derivatives, analogues, isomers and salts thereof
Figure imgf000079_0001
Formula 1
Wherein n= 1 or 2, dotted lines indicates the single or double bond, wherein ri=l the substituent R7 at C-2 position is H radical, whereas when n=2, the substituent R7 is at C-4 position and R7 is a normal or branched chain C1 to Ci0 alkyl group, phenyl, benzyl radical and the like, wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Ci to Cio) or aryl or benzyl (unsubstituted and substituted);
Figure imgf000079_0002
is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, substituted pyranyl radical, unsubstituted pyranyl radical or Ri1 R4 represents hydrogen group and R2 ,R3 represent hydrogen , alkoxyl (Cl- ClO), alkenoxyl groups (Ci-C10), aryloxyl, substituted aryloxyl groups or Ri1 R4 represents hydrogen group and R2 + R3 together represent 1,3-dioxol , 1,4-dioxol groups ; R5 represents hydrogen atom or normal or branched chain Ci to Ci0 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain Ci to Ci0 alkyl group or phenyl or benzyl radical; where NR5R6 together (R5+Re) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N- methylpiperazinyl, oxazolyl, N-methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl
2. A compound as claimed in claim 1 wherein the compound of the general formula 1 is substituted 3 -(unsubstituted or substituted 3,4-dihydronaphth-2-yl)- acrylic acid amide of formula Ia
Figure imgf000080_0001
formula-la
Wherein R represents halogen or hydrogen, alkyl (C1 to C1O ) , aryl, or benzyl group (substituted or un-substituted), R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, substituted pyranyl radical, unsubstituted pyranyl radical, or R1, R4 represents hydrogen group and R2 , R3 represent hydrogen ,alkoxyl (Cl-ClO) , alkenoxyl groups (C1-C1O), aryloxyl substituted aryloxyl groups or , R1, R4 represents hydrogen group and R2 + R3 together represent
1,3-dioxol ,1,4-dioxol groups ; R5 represents hydrogen atom or normal or branched chain C1 to Ci o alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain Ci to Ci0 alkyl group or phenyl or benzyl radical; wherein NR5R6 together (R5+R6) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl, N-methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl.
A compound as claimed in claim 1 wherein the derivative of compound of general formula 1 is substituted dihydronaphthalene pentadienoic acid amide of formula Ib;
Figure imgf000080_0002
Formula Ib
Wherein R represents halogen or hydrogen, alkyl (Ci to Cio), aryl, or benzyl group (substituted or un-substituted), R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, substituted pyranyl radical , unsubstituted pyranyl radical or Ri, R4 represents hydrogen group and R2, R3 represent hydrogen, alkoxyl(Cl-ClO) , alkenoxyl groups (Ci -Ci0), aryloxyl,/substituted aryloxyl groups or Ri1 R4 represents hydrogen group and R2 + R3 together represent 1,3-dioxol, 1,4- dioxol groups ; R5 represents hydrogen atom or normal or branched chain Ci to C10 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain C1 to C10 alkyl group or phenyl or benzyl radical; where NR5R6 together (R5+R6) represents heterocyclic amine radical such as piperidinyl, pyrrαlidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl N-
10 methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like and optionally converting them to their salts by method known in the art of synthesis; where R7 represents normal or branched chain Cj to C10 alkyl group or phenyl or " benzyl radical.
L5
A compound as claimed in claim 1 wherein tetrahydroderivative of compound of general formula 1 is having structural formula Iaa
Figure imgf000081_0001
Formula Iaa
»0
Wherein R represents halogen or hydrogen, alkyl (Ci to Cio ) > aryl, or benzyl group (substituted or un-substituted), R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, substituted pyranyl radical , unsubstituted pyranyl radical or Ri, R4 represents hydrogen group and R2 , R3 represent hydrogen ,
!5 alkoxyl(Cl-ClO) , alkenoxyl groups (CpCio) , aryloxyl, substituted aryloxyl groups or Ri. R4 represents hydrogen group and R2 + R3 together represent 1,3-dioxol ,1,4- dioxol ; R5 represents hydrogen atom or normal or branched chain C1 to C)0 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain C1 to Ci0 alkyl group or phenyl or benzyl radical; wherein NRjR6 together (R5+R6) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl., morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl, N- methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyL •
compound as claimed in claim 1 wherein the hexahydroderivative of general formula 1 is having structural formula lba
Figure imgf000082_0001
Formula lba
Wherein R represents halogen, or hydrogen, alkyl (C1 to Cio ) , aryl, or benzyl group (substituted or un-substituted), R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, substituted pyranyl radical , unsubstituted pyranyl radical or R1, R4 represents hydrogen group and R2 , R3 represent hydrogen, alkoxyl(Cl to ClO) , alkenoxyl groups (Ci-Ci0), aryloxyl, substituted aryloxyl groups or R^R4 represents hydrogen group and R2 + R3 together represent 1,3- dioxol, 1,4-dioxol ; R5 represents hydrogen atom or normal or branched chain Ci to Ci0 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain Ci to Cio alkyl group or phenyl or benzyl radical; where NR5R6 together (R5+R6) represents heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl N- methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl ■ wherein R7 represents normal or branched chain Ci to Cio alkyl group or phenyl or benzyl radical.
A compound as claimed in claims 1-5 wherein the representative compounds of general formula 1 comprising the compounds; 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid piperidide, 3 -(I -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid pyrrolidide, 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid morpholide, 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid n-octylamide, 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide, 3-(l-chloro-3,4-dihydiOnapth-2-yl)-acrylic acid isopropylamide, 3-(l-chloro-3,4-dihydronapth-2-yl)-acrylic acid N,N-diethylamide, 3 -(I -chloro-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisopropylamide, 3 -( 1 -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid p-methoxyanilide, -(I -chloro-3,4-dihydronapth-2-yl)-acrylic acid 3,4-methylenedioxyanilide, 3-(l -chloro-3 ,4-dihydro-napth-2-yl)-acrylic acid o-methylanilide, -(l -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid benzylamide, -( 1 -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid ^-nitroanilide, -(l-chloro-3,4-dihydro-napth-2-yl)-acrylic acid />-fiuoroanilide, -(I -chloro-3 ,4-dihydronaρth~2-y.i)-acrylic acidj^-carboxyanilide, -(l -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid N,N-diphenylamide, -(I -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid N-methylpiperazide, -( 1 -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid ;?-methylanilide, -(I -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid «-butylamide, -(I -chloro-3, 4-dihydronapth-2-yl)-acrylic acid anilide, -(I -chloro-3, 4-dihydronapth-2-yl)-acrylic acid imidazolide, -(I -chloro-3, 4-dihydronapth-2-yl)-acrylic acid ø-methoxyanilide, -(l -chloro-3 ,4-dihydronapth-2-yl)-acrylic acid N,N-diisobutylamide, -(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid piperidide, • -(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid pyrrolidide, -(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid morpholide, -(l-bromo-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide, 3 -(I -bromo-3,4-dihydronapth-2-yl)-acrylic acid N,N-diisopropylamide, 3-(l-bromo-3,4-dihydro-napth-2-yl)-acrylic acid fl-butylamide, 3-(l-bromo-3,4- dihydronaρth-2-yl)-acrylic acid N-methylpiperazide, 3 -(I -bromo-3,4-dihydronapth-2-yl)-acrylic acid N,N-diethylamide, 3 -( 1 -bromo-3 ,4-dihydro-napth-2-yl)-acrylic acid imidazolide,
3 -( 1 -bromo-3 ,4-dihydronapth-2-yl)-acrylic acid />-hydroxypiperidide, 3-(3,4-dihydronaphth-2-yl)-acrylic acid isobutylamide, 3-(3,4-dihydronaphtli-2-yl)-acrylic acid diisopropylamide, 3-(3,4-dihydronaphth-2-yl)-acrylic acid piperidide, 3 -(3 ,4-dihydronaphth-2-yl)-acrylic acid pyrrolidide,
3-(3,4-dihydronaρhth-2-yl)-acrylic acid morpholidide, 3-(3,4-dihydronaphth-2-yl)-acrylic acid/>-methoxy anilide,
3-(3,4-dihydronaphth-2-yl)-acrylic acid octylamide, 3-(3,4-dihydronaphth-2-yl)- acrylic acid diethylamide, 3-(3,4-dihydronaphth-2-yl)-acrylic acid anilide,
3-(3}4-dihydronaphth-2-yl)-acrylic acid 3,4-methylenedioxy anilide, 3-(l-benzyl-3,4-dihydro-napth-2-yl)-acrylic acid «-octylamide, 3-(l-benzyl-3,4-dihydro-napth-2-yl)-acrylic acid piperidide, 3-(l-benzyl-3,4-dihydro-napth-2-yl)-acrylic acid pyrrolidide, 3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid morpholide,
3-(l-benzyl-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide,
3 -( 1 -benzyl-3 ,4-dihydronapth-2-yl)-acrylic acid N,N-diisoprόpylamide,
3 -( 1 -benzyl-3 ,4-dihydronapth-2-yl)-acrylic acid jo-hydroxy piperidine,
3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid piperidide, 3-(6,7- dimethoxy-3,4-dihydronaρhth-2-yl)-propenoic acid morpholide,
3 -(6,7-dimethoxy-3 ,4-dihydronaphth-2-yl)-propenoic acid pyrrolidide, 3 -(6,7-dimethoxy-3 ,4-dihydronaphth-2-yl)-propenoic acid w-octylamide, 3-(657-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid isobutylamide, 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid «-butylamide, 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid π-propylamide,
3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid isopiOpylamide, 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid N,N-diisopropylamide, 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid anilide, 3-'(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic-acid-o-methylanilide, 3 -(6,7-dimethoxy-3 ,4-dihydronaphth-2-yl)-propenoic acid /?-methylanilide, 3-(6,7-dimethoxy-3,4-dihydronaphth-2-yl)-propenoic acid£>-methoxyanilide,
3-(l-methyl-3,4-dihydronapth-2-yl)-acrylic acid piperidide, 3-(l-methyl-3,4-dihydronapth-2-yl)-acrylic acid pyrrolidide, 3-(l-methyl-3,4-dihydronapth-2-yl)-acrylic acid morpholide, 3-(l-methyl-3,4-dihydronapth-2-yl)-acrylic acid isobutylamide, 3 -( 1 -methyl-3 ,4-dihydronapth-2-y l)-acrylic acid N,N-diisopropylamide.
7. A compound of the formula 1 as claimed in claims 1 wherein the analogs of compound of general formula 1 are la,laa,lb,lba.
8. A compound of the formula 1 as claimed in 1 wherein the salt is pharmaceutically acceptable selected from a group consisting of hydrochloride, acetate, succinate, maleate.
9. A compound of the formula 1 as claimed in claims 1-8 are useful as potentiators of antiinfective drugs.
10. A compound of the formula 1 as claimed in claims 1 wherein the ED50 of antiinfective drug is reduced to about 1/4 when used in combination with the compound of formula Ia, laa, Ib and lba.
11. A compound of the formula 1 as claimed in claims 1 wherein the dose of antiinfective drugs is reduced about 1A when used in combination with compounds of formula la,laa,lb and lba.
12. A process for preparation of compound of general formula 1 as claimed in claim 1 the process comprising the steps; (i) subjecting aldehyde compound of formula 2,
Figure imgf000086_0001
wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Ci to Cio) or aryl or benzyl (unsubstituted and substituted); R1, R2; R3, R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, substituted pyranyl radical, unsubstituted pyranyl radical or R1, R4 represents hydrogen group and R2, R3 represent hydrogen, alkoxyl (Cl -ClO), alkenoxyl groups (C1-C1O) , aryloxyl , substituted aryloxyl groups or Ri1 R4 represents hydrogen group and R2 + R3 together represent 1,3- dioxol,l,4-dioxol, to Wittig reaction (Wittig reagent prepared from triphenylphosphine and ethyl bromoacetate/ ethylchloroacetate in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at temperature 5-80 0C for 1 to 24 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like benzene/toluene or dimethylformamide) and subsequent saponification of the Wittig reaction product to get compound of the formula 3,
(ii) alternately allowing aldehyde of the formula 2 to react with malonic acid in presence of a base like piperdine and pyridine, contents stirred at temperature 25-40 0C for 24-30 hours followed by subsequent decarboxylation carried out by heating at 80-90 0C, acidification of the reaction mixture by dilute HCl, or H2SO4 or the like, extraction of the contents, concentration of the organic layer, to get the compound of formula 3,
(iii) treating the compound of formula 3 with thionylchloride and subsequent reaction of the acid chloride thus prepared with an appropriate amine to get the compound of formula Ia by conventional method wherein RiR2R3R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, substituted and unsubstituted pyranyl radcial or Ri, R4 represents hydrogen group and R2, R3 represent hydrogen , alkoxyl (Cl to ClO) , alkenoxyl groups (Ci-Ci0) , aryloxyl , substituted aryloxyl groups or R1, R4 represents hydrogen group and R2 + R3 together represent 1,3-dioxol or 1,4- dioxol; R, represents hydrogen atom or normal or branched chain C1 to Ci0 alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain Ci to Cj0 alkyl group or phenyl or benzyl radical; where NR5R6 together (R5+R6) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl, N- methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and optionally converting them to their salts by known method by reacting with an' acidic compounds such as hydrochloric acid, acetic acid, tartaric acid,
Figure imgf000087_0001
Formula 1 a
(iv) converting the compound of formula Ia to its tetrahydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol at ambient temperature and at 1-3 atmospheric pressure to obtain corresponding tetrahydro derivative of general formula laa,
Figure imgf000087_0002
Formula laa
(v) alternately, hydrogenation of compounds of the formula 3 with Pd/charcoal/ solvents such as methanol, ethanol, ethylacetate and at 1-3 atmospheric pressure and treatment of the tetrahydro derivative with thionyl chloride and followed by treatment of the acid chloride with appropriate amine afforded compound of the formula laa wherein R,R1,R2,R3,R4,R5,R6 are radicals/groups as described above for the compound of the formula Ia,
(vi) For the preparation of compound of the formula Ib the method of preparation comprises of reacting compound of the formula 2 with Grignard reagent prepared from appropriate alkyl halide (represented by R7X, chosen from Cl-ClO normal of branched alkylhalide, substituted or unsubstituted benzyl or aryl halides) and Mg metal with constant stirring at ambient temperature in an anhydrous ether/THF solvent to produce compound of the formula 4 wherein R1JRa1R31R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, substituted pyranyl radical , unsubstituted pyranyl radical or R1, R4 represents hydrogen group and R2,R3 represent hydrogen, alkoxyl(Cl-ClO), alkenoxyl groups (Ci-C10) ,aryloxyl , substituted aryloxyl 'groups or R1, R4 represents hydrogen group and R2+R3 together represent 1,3-dioxol , 1,4- dioxol ;R7 is Cl-ClO normal or branched allcylhalide, substituted or unsubstituted benzyl or aryl halides,
Figure imgf000088_0001
Formula Ib
(vii) treating the compounds of formula 4 with Vilsmeier reagent comprising of dimethylformamide and phosphorus oxychloride/phosphorus oxybromide or phosphorus tribromide mixture at 0-50 0C preferably at 0-10 0C for 5-50 hrs. Preferably for 5-12 hrs, followed by dilution of contents with ice cold water, bringing the pH to 7 by neutralisation with dilute alkali solution to produce compound of formula 5 wherein R1R2R3R4 and R7 are radicals or substituents as described in for compounds of the formula 4,
Figure imgf000088_0002
Figure imgf000088_0003
Formula 5
(viϋ) condensation of the compounds of the formula 5 with an ylide (Wittig reagent, prepared from triphenylphosphine and ethyl, bromoacetate/ ethylchloroacetate) in equimolar mixture in presence of a strong base such as sodium hydride, sodium methoxide and the like at a temperature ranging between 5-80 0C for a period ranging between 1 to 24 hrs in an ethereal medium such as diethyl ether, dimethoxyethane and the like," or benzene/toluene or dimethylformamide followed by direct saponification of Wittig reaction product furnished compounds of the formula 6 wherein R1R2R3R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, substituted and unsubstituted pyranyl radcial or R1, R4 represents hydrogen group and R25R3 represent hydrogen, alkoxyl (C1-CiO) » alkenoxyl groups (C1-C10) , aroyloxyl , substituted aroyloxyl groups or Rj s ' R4 represents hydrogen group and R2+R3 together represent 1,3-dioxol or 1,4-dioxol ;R7 is Cl-ClO normal or branched alkylhalide, substituted or unsubstituted benzyl or aryl radical,
Figure imgf000089_0001
Formula θ
(ix) treating the compound of the general formula 6 with thionyl chloride in presence of a base such as pyridine followed by treatment of the acid chloride with an appropriate base afforded compounds of the formula Ib wherein R is selected from a group consisting of halogen, hydrogen, alkyl (Cl to ClO) or aryl or benzyl (unsubstituted and substituted); R1,R2,R3,R4 is hydrogen, nitro, halogen, hydroxy, alkyloxy, alkenyloxy, aryloxy, substituted pyranyl radical,unsubstituted pyranyl radical or Rl, R4 represents hydrogen group and R2, R3 represent hydrogen, alkoxyl(Ci to ClO) , alkenoxyl groups (Cl-ClO), aryloxyl, substituted aryloxyl groups or Rl, R4 represents hydrogen group and R2 + R3 together represents 1,3- dioxol or 1,4-dioxol,; R5 represents hydrogen atom or normal or branched chain Ci to Cio alkyl group or phenyl or benzyl radical and R6 represents hydrogen atom or normal or branched chain Ci to Ci0 alkyl group or phenyl or benzyl radical; where NR5R6 together (Rj+Re) represent heterocyclic amine radical such as piperidinyl, pyrrolidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, oxazolyl, N- methylpiperazinyl, pyrrolyl, imidazolyl, oxazolyl or an amino acid such as alaninyl, leucinyl, phenylalaninyl, tyrosinyl, glycylglycinyl, alanylalaninyl and the like, wherein R7 represents normal or branched chain Cl to ClO alkyl group,
Figure imgf000090_0001
Formula Ib
(x) converting the compound of formula Ib to its tetra hydro derivative by hydrogenation in presence of Pd/charcoal in polar solvent such as methanol or ethanol or ethyl acetate at ambient temperature and at a pressure ranging between 1- 3 atmosphere to obtain compound of formula lba.
Figure imgf000090_0002
Formula lba
(xi) alternately, hydrogenation of compounds of the formula 6 with Pd/charcoal/ solvents such as methanol, ethanol, ethyl acetate and at a pressure ranging between 1-3 atmosphere and subsequent treatment of the tetrahydro derivative with thionyl chloride followed by treatment of the acid chloride thus obtained with appropriate amine afforded compound of the formula lba wherein RR1R2R3R4R5R6R7 are radicals/groups as described above for the compound of the formula Ib.
13. A pharmaceutical composition comprising one or more of compound of the formula 1 as claimed in claim 1, an anti-infective compound, optionally along with an excipient or pharmaceutical vehicle.
14. A pharmaceutical composition as claimed in claim 13 wherein the amount of compound of formula Ia, laa,lb, lba as claimed in claims 1-9, in the composition may vary from 0.1 to 50% by weight of the drug and the amount of the drug may vary from 50 to 99.9% by weight of the composition.
15. A composition as claimed in claim 13 the composition is useful as anti-microbial agent.
5 16. A composition as claimed in claim 13 wherein the minimum inhibitory concentration (MIC) value of anti-infective drugs in the composition is reduced by more than 32 times of the concentration when used alone.
17. A pharmaceutical composition as claimed in claim 13 wherein the anti-infectives are 10 selected from a group (but not limited to) comprising of penicillins (including semisynthetic), cephalosporins, aminoglycosides, glycopeptides, fluroquinolones, macrolides, tetracyclines other antibiotic groups such as mupirocin and framycetin, first and second line anti-TB drugs, anti-leprosy drugs, oxazolidinones.
15 18. A pharmaceutical composition as claimed in claim 13 , wherein the said composition along with excipient or pharmaceutical vehicle selected from a group consisting of lactose, corn starch, polyethylene glycols may be administered through oral route.
19. A pharmaceutical composition as claimed in claim 13 , wherein the said composition 20 along with excipient or pharmaceutical vehicle selected from a group consisting of castor oil, olive oil may be administered through systemic route.
20. A pharmaceutical composition as claimed in claim 13 , wherein the said composition along with excipient or pharmaceutical vehicle selected from a group consisting of
15 polyethylene glycols, bees wax, paraffin wax, emulsifying agents may be applied topically.
21. A pharmaceutical composition as claimed in claim 13 wherein composition is effective against microorganisms selected from Mycobacterium species, Gram-
)0 positive bacteria such as Staphylococcus species and Bacillus species, Gram-negative bacteria such as Pseudomonas species, E. coli and Salmonella species.
22. A pharmaceutical composition as claimed in claim 13 wherein composition is effective against growth of microorganisms including resistant strains such as methillin resistant Staphylococcus aureus (MRSA).
23. A pharmaceutical composition as claimed in claim 13 , wherein composition is effective when tested for curing of mice, guinea pig or rabbit models infected with microorganisms from the group of bacteria such as Staphylococcus species and Bacillus species, Pseudomonas species, E coli and Salmonella species, Mycobacterium species.
24. A method for inhibiting a bacterial cell that employs an efflux pump resistance mechanism, comprising contacting the cell with a pharmaceutical composition comprising of Iba, laa, Ib, lba as claimed in claim 1-9 and optionally along with excipient or pharmaceutical vehicle.
25. A method of claim 22 wherein bacterial cell is selected from Gram-positive bacteria, Gram- negative bacteria or mycobacterial species such as Staphylococcus species, Bacillus species, Pseudomonas species, E coli and Salmonella specie. '
26. A method of treating infections comprising administrating to a subject in need of such treatment a therapeutically effective amount of the composition comprising one or more of compound of the formula 1 as claimed in claim 1, an anti-infective compound, optionally along with an excipient or pharmaceutical vehicle.
27. A method for treating an infection caused by a bacteria that employs an efflux pump resistance as one of the ways of resistance, comprising administering to a patient in need thereof a therapeutically effective amount of an antibacterial agent and a compound of formula Ia, laa, Ib, lba as claimed in claims 1-9.
8. Novel efflux pump inhibitors, process of preparation, a pharmaceutical composition thereof substantially as herein described with reference to the examples and drawings accompanying the specification.
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