Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
  • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
  • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom 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
  • C07D215/48—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
• • 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
  • A61P31/12—Antivirals
  • A61P31/14—Antivirals for RNA viruses
• • 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
  • A61P31/12—Antivirals
  • A61P31/14—Antivirals for RNA viruses
  • A61P31/18—Antivirals for RNA viruses for HIV
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• the subject of the invention is quinoline derivatives having in particular properties which inhibit retrovirus integrases. It also relates to a process for the synthesis of these derivatives and to the drugs containing them in their active principle.
• the object of the invention is therefore to provide new quinoline derivatives endowed in particular with retrovirus anti-integrase properties.
• the invention also relates to pharmaceutical compositions containing these derivatives.
• the quinoline derivatives according to the invention are characterized in that they correspond to formula I
• X represents an alkyl chain- (CH 2 ) n - in which n is equal to 0, 1 or 2, O, or N,
• Z represents an aromatic ring which may contain heteroatoms chosen from O, N or S, in substitution for the carbon atoms constituting said aromatic ring, this cycle may or may not be substituted by Rb,
• - Rb represents 1 to 3 identical or different substituents, chosen from the groups -OH, -OR, -COOH, -COOR, -COH, -COR, -NH 2 , -NH (R), -NH (R, R ' ), -SH and -SR and CN,
• - Ra represents a hydrogen atom or a group - (CH 2 ) n.
• -Y for which n 'is equal to 0, 1, 2 or 3 and Y represents -CH 3 , -COOH, -COOR, -CN , -OH, -OR, SR, or an aryl group optionally substituted by Rb, - R and R ', identical or different, represent a linear or branched alkyl chain of 1 to 4 carbon atoms, and
• Preferred derivatives are 7-carboxy-8-hydroxyquinolines, and their pharmaceutically acceptable salts, chosen from 2- (3, 4-dihydroxy-benzylcarbamoyl) - 8-hydroxy- quinoline-7-carboxylic, 2- (2,4-dihydroxy-benzylcarbamoyl) -8-hydroxy-quinoline-7-carboxylic acid and their sodium salts.
• the invention also relates to a process for the synthesis of the derivatives defined above.
• R-_ and R 2 identical or different, represent alkyl radicals from 1 to 4C, with an amine of formula III
• the condensation of the succinimidyl quinolinecarboxylate of formula II with the amine salt is carried out in pyridine at room temperature. By operating at approximately 20 ° C., the reaction time is of the order of 10 to 15 h.
• the amine salt is in particular a para-toluenesulfonate
• the amide IA is then obtained by saponification, followed by acidification.
• the saponification is carried out for example with a 3N sodium hydroxide solution. This reaction can be carried out in methanol at room temperature for approximately 2 to 4 hours, in particular 2 hours.
• For acidification use is made in particular of dilute sulfuric acid.
• succinimidyl quinolinecarboxylate derivative of formula II is reacted with an amine of the following formula V or its organic salt:
• succinimidyl quinolinecarboxylate of formula is reacted. II with the hydrazine of formula VI below:
• succinimidyl quinolinecarboxylate of formula II is obtained from 8-hydroxy-7-quinaldine carboxylic acid of formula VIII:
• the protection of the -COOH and -OH groups is carried out using derivatives capable of introducing specific protective groups with respect to the functions concerned and which can then be selectively cut without altering the rest of the molecule.
• esterification of the -COOH group will be carried out first, followed by that of the -OH group.
• the first esterification reaction is carried out at a temperature of the order of 100 ° C., for approximately 50 to 80 h, in particular for approximately 70 h.
• the treatment of the ester obtained with an acyl halide is carried out at ambient temperature in the presence of pyridine, for approximately 30 min to 2 h, and in particular for 1 h.
• the diester derivative obtained is oxidized in two stages.
• the -CH 3 group is oxidized to C-2 of the quinoline to the -COH group, by treatment with an oxidizing agent of -CH 3 group such as selenium oxide.
• an oxidizing agent of -CH 3 group such as selenium oxide.
• a solvent such as dioxane is refluxed for approximately 10 to 15 hours, in particular 12 hours, in order to obtain the corresponding aldehyde.
• this aldehyde is then oxidized to acid.
• Suitable oxidizing agents include sodium chlorite.
• the group -COOH is activated. using N-hydroxy-succinimide.
• the condensation of the acid with N-hydroxy-succinimide is advantageously carried out in the presence of dicyclohexylcarbodiimide. By operating at room temperature, of the order of 20 ° C., the reaction lasts for approximately 10 to 15 hours, in particular 12 hours.
• the invention therefore relates to pharmaceutical compositions characterized in that they comprise an effective amount of at least one derivative as defined above, in association with pharmaceutically acceptable vehicles.
• compositions are advantageously used in combination with other anti-HIV drugs, in particular drugs with an inhibitory effect with respect to reverse transcriptase and / or protease.
• the dosages and modes of administration will be adapted according to the treatment with mono or polytherapy used.
• the invention also relates to the use of the derivatives defined above as biological reagents which can be used in particular for studies of mechanisms relating to viral infection.
• FIGS. 1 to 3 show, respectively:
• FIGS. 2A and 2B their absence of toxicity on adherent and suspended cells, and - Figures 3A and 3B, the inhibition of all of the integrated viral DNA (Figure 3A) and the detection of total viral DNA ( Figure 3B).
• the hydroxy acid 1 previously described in literature 1 is treated at 100 ° C. for 72 hours with n-butanol in the presence of polyphosphoric acid to provide the butyl ester 2.
• This ester treated with pivaloyl chloride in pyridine for 1 hour at 20 ° C. gives the diester 3.
• This is oxidized by means of selenium dioxide at the reflux of dioxane for 12 hours to give the aldehyde 4, which is oxidized to acid 5 by means of sodium chlorite in a water ⁇ er ⁇ -butanol mixture for 12 hours, in the presence of sodium dihydrogenophosphate and 2-methyl-2-butene.
• the succinimidyl quinoline carboxylate 6 is prepared by condensing the acid 5 on the N-hydroxysuccinimide in the presence of dicyclohexylcarbodiimide for 12 hours at 20 ° C.
• Step 3 preparation of 8- ( 2,2 -dimethyl-propionyloxy) -2-formyl-quinoline-7-n-butyl carboxylate (4)
• a mixture of the preceding diester 3 (step 2) (11.5 g, 34 mmol) and selenium dioxide (6.7 g, 60 mmol) in 50 ml of 1,4-dioxane is brought to reflux for 12 hours. After cooling to 20 ° C, the reaction mixture is filtered through Cêlite® and concentrated under reduced pressure.
• step 3 A mixture of the above aldehyde 4 (step 3) (3.5 g, 10 mmol) of sodium dihydrogenophosphate (1.7 g, 12 mmol), of 2-methyl-2-butene (6.1 ml, 57 mmol) and sodium chlorite (2.6 g, 29 mmol) is stirred at 20 ° C in 160 ml of ter-butanol / water mixture 5/1 and 30 ml of dichloromethane for 12 hours. The reaction medium is concentrated under reduced pressure.
• step 4 A mixture of the previous acid 5 (step 4) (1.2 g, 3.2 mmol), dicyclohexylcarbodiimide (0.7 g, 3.2 mmol) and N-hydroxysuccinimide (0.4 g, 3.2 mmol) in 12 ml of dichloromethane is stirred at 20 'C for 12 hours. The reaction medium is concentrated under reduced pressure.
• step 5 A mixture of the preceding product 6 (step 5) (1.4 g, 2.9 mmol) and the paratoluenesulfonate of 3,4-dihydroxybenzylamine (0.9 g, 2.9 mmol) in 10 ml of pyridine is stirred at 20 ° C for 12 hours. The mixture is concentrated under reduced pressure, and 10 ml of water are added. The mixture is acidified to pH 4 with a 2N hydrochloric acid solution, then 20 ml of dichloromethane are added. The phases are separated and the aqueous phase extracted with dichloromethane. The combined organic phases are dried over magnesium sulfate and concentrated under reduced pressure.
• step 6 A mixture of the preceding amide 7 (step 6) (0.5 g, 1.0 mmol) and 6.5 ml of a solution of 3 N sodium hydroxide in 10 ml of methanol is stirred at 20 ° C for 3 hours. The pH is adjusted to 4 with a 1N sulfuric acid solution. The reaction mixture is concentrated under reduced pressure, then dried under vacuum over phosphoric anhydride. The residue is taken up in 10 ml of hot anhydrous ethanol and the insoluble sodium sulfate is filtered. The filtrate is concentrated under reduced pressure, then taken up in 2 ml of anhydrous ethanol. The medium is cooled: a yellowish solid crystallizes. The latter is washed with dichloromethane, then dried under vacuum over phosphoric anhydride to give 250 mg of acid 8 in the form of a brown solid.
• step 7 A mixture of the preceding amide 8 (step 7) (20 mg, 0.06 mmol) and 0.66 ml of a 0.1 N sodium hydroxide solution are stirred at 20 ° C for 10 minutes. The sodium salt obtained is lyophilized to quantitatively give 22 mg of dark brown solid.
• HIV-1 integrase activity is measured by the following tests:
• the endonucledlytic activity of the recombinant protein is tested on a double stranded oligonucleotide of 21 base pairs; radiolabelled at the 5 'end. Integrase activity results in the elimination of the dinucleotide at the 3 'end.
• the strand transfer test is carried out with a double-stranded oligonucleotide of 21 base pairs mimicking the end of the viral DNA from which the 3 ′ dinucleotide has been deleted.
• the activity of the protein results in the covalent insertion of this oligonucleotide into a homologous oligonucleotide.
• the disintegration test is carried out with a substrate mimicking the structure of the integrated viral DNA.
• the amount of DNA excised by the integrase is measured. This last test only measures the catalytic activity of the protein, excluding its DNA binding activity.
• the enzymatic activities are tested in the presence of manganese and / or magnesium.
• the compounds according to the present invention inhibit the enzymatic activities, both in the presence of manganese and of magnesium.
• the first test consists in bringing cells of an established lymphocyte line, CEM cells, into contact with a supernatant of infected cells containing the infectious virions.
• the test is carried out as follows: CEM cells, cultured in suspension in RPMI medium supplemented with 10% fetal calf serum, are infected with a viral supernatant with a multiplicity of infection of 0.5. After two hours of infection, the cells are washed twice with RPMI so as to remove the residual viral particles. The cells are then replaced in RPMI medium containing the compound according to the invention.
• the viral load is evaluated after 72 hours of culture. This is quantified by the following two methods:
• the amount of p24 viral protein is determined by an ELISA test.
• the load of infectious virus is estimated by infection of the HeLa ⁇ -gal CD4 + cells.
• the efficiency of retroviral integration is estimated by a colorimetric test (CPRG).
• the second test consists in bringing HeLa ⁇ -gal CD4 + cells (human fibroblastic cells containing the ⁇ -galactosidase gene under transcriptional control of the viral transactivator Tar and expressing the CD4 membrane receptor) into contact with the supernatant of cells transfected by the plasmid.
• HeLa ⁇ -gal CD4 + cells human fibroblastic cells containing the ⁇ -galactosidase gene under transcriptional control of the viral transactivator Tar and expressing the CD4 membrane receptor
• HeLa ⁇ -gal CD4 + are cultured in 96-well plates in DMEM medium supplemented with 10% fetal calf serum, brought into contact with 5 ng of p24 (derived from a transfection of 293T cells with the plasmid pNL43) and dilutions of the compound according to the invention. After 48 hours, the cells are lysed and the ⁇ -galactosidase is assayed by staining with CPRG.
• the compounds according to the invention show a protective effect against infection by the HIV-1 virus. Indeed, this protective effect results in an inhibition of the production of viral particles in the first test and an inhibition of CPRG staining in the second test, at micromolar or even submicromolar concentrations.
• the IC50s vary from 0.3 ⁇ M to 4 ⁇ M.
• the compound tested is moreover devoid of cytotoxicity at 500 ⁇ M both on adherent cells (HeLa) and in suspension (CEM) as illustrated by the results reported in FIGS. 2A (- * - CEM; - U— HeLa ) and 2B (- * - BioA53; - M— BioA162 - - BioA163).
• This test consists in bringing CEM cells into contact with the supernatant of infected cells containing the infectious virions. After 2 hours, the cells are washed and replaced in RPMI medium containing the compound according to the invention for 24 hours (BioA53).
• the DNA of the cells is then extracted using a genomic DNA preparation kit (QiaBlood, Qiagen ' ).
• This DNA is the target of PCR amplifications aimed at evaluating, firstly, the quantity of total viral DNA present in the solution, and secondly, the quantity of viral DNA integrated into the cellular genome.
• This last amplification takes place in two stages: the first step uses a PCR primer homologous to a retroviral DNA sequence; the second primer is homologous to the Alu repeat genomic sequences.
• the second step is a nested PCR to reveal the retroviral DNA.
• the compound according to the present invention shows a decrease of at least 80% in the amount of integrated DNA for a concentration of 50 ⁇ M and an inhibition of all of the integrated DNA at 100 ⁇ M (FIG. 3A) while the amount of Total viral DNA is not affected by treatment of cells with the compound (Figure 3B).

Landscapes

• Health & Medical Sciences (AREA)
• Organic Chemistry (AREA)
• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Virology (AREA)
• Veterinary Medicine (AREA)
• Pharmacology & Pharmacy (AREA)
• Public Health (AREA)
• General Health & Medical Sciences (AREA)
• Animal Behavior & Ethology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Medicinal Chemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Molecular Biology (AREA)
• Oncology (AREA)
• Communicable Diseases (AREA)
• Tropical Medicine & Parasitology (AREA)
• AIDS & HIV (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Quinoline Compounds (AREA)
• Plural Heterocyclic Compounds (AREA)
• Nitrogen Condensed Heterocyclic Rings (AREA)

Priority Applications (9)

Applications Claiming Priority (2)

Related Child Applications (1)

Publications (1)

Family

ID=8868255

Family Applications (1)

Country Status (10)

Cited By (4)

Families Citing this family (2)

Citations (1)

Family Cites Families (1)

• 2001
  • 2001-10-12 FR FR0113209A patent/FR2830863B1/fr not_active Expired - Fee Related
• 2002
  • 2002-10-14 IL IL16132002A patent/IL161320A0/xx unknown
  • 2002-10-14 WO PCT/FR2002/003512 patent/WO2003031413A1/fr not_active Ceased
  • 2002-10-14 DK DK02795314T patent/DK1461319T3/da active
  • 2002-10-14 ES ES02795314T patent/ES2326115T3/es not_active Expired - Lifetime
  • 2002-10-14 AT AT02795314T patent/ATE430134T1/de not_active IP Right Cessation
  • 2002-10-14 JP JP2003534397A patent/JP2005508350A/ja active Pending
  • 2002-10-14 EP EP02795314A patent/EP1461319B1/fr not_active Expired - Lifetime
  • 2002-10-14 US US10/492,088 patent/US7064133B2/en not_active Expired - Fee Related
  • 2002-10-14 DE DE60232186T patent/DE60232186D1/de not_active Expired - Lifetime
• 2004
  • 2004-04-07 IL IL161320A patent/IL161320A/en not_active IP Right Cessation
• 2006
  • 2006-03-01 US US11/364,329 patent/US20060148849A1/en not_active Abandoned

Patent Citations (2)

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events