Classifications

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  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
  • C07F9/655—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms
  • C07F9/65502—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms the oxygen atom being part of a three-membered ring
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/66—Phosphorus compounds
• • 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
• • 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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/02—Immunomodulators
  • A61P37/04—Immunostimulants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/02—Immunomodulators
  • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/08—Antiallergic agents
• • 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
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/06—Phosphorus compounds without P—C bonds
  • C07F9/22—Amides of acids of phosphorus
  • C07F9/24—Esteramides
  • C07F9/2404—Esteramides the ester moiety containing a substituent or a structure which is considered as characteristic
  • C07F9/2425—Esteramides the ester moiety containing a substituent or a structure which is considered as characteristic containing the structure (RX)(RR'N)P(=Y)-Z-(C)n-Z'-P(=Y)(XR)2 (X = O, S, NR; Y = O, S, electron pair; Z = O, S; Z' = O, S)
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/06—Phosphorus compounds without P—C bonds
  • C07F9/22—Amides of acids of phosphorus
  • C07F9/24—Esteramides
  • C07F9/2404—Esteramides the ester moiety containing a substituent or a structure which is considered as characteristic
  • C07F9/2433—Compounds containing the structure N-P(=X)n-X-acyl, N-P(=X)n-X-heteroatom, N-P(=X)n-X-CN (X = O, S, Se; n = 0, 1)
  • C07F9/245—Compounds containing the structure N-P(=X)n-X-acyl, N-P(=X)n-X-heteroatom, N-P(=X)n-X-CN (X = O, S, Se; n = 0, 1) containing the structure N-P(=X)n-X-P (X = O, S, Se; n = 0, 1)
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/06—Phosphorus compounds without P—C bonds
  • C07F9/22—Amides of acids of phosphorus
  • C07F9/24—Esteramides
  • C07F9/2454—Esteramides the amide moiety containing a substituent or a structure which is considered as characteristic
  • C07F9/2458—Esteramides the amide moiety containing a substituent or a structure which is considered as characteristic of aliphatic amines
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/06—Phosphorus compounds without P—C bonds
  • C07F9/22—Amides of acids of phosphorus
  • C07F9/24—Esteramides
  • C07F9/2454—Esteramides the amide moiety containing a substituent or a structure which is considered as characteristic
  • C07F9/2462—Esteramides the amide moiety containing a substituent or a structure which is considered as characteristic of unsaturated acyclic amines
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
  • C07F9/655—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms
  • C07F9/65502—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms the oxygen atom being part of a three-membered ring
  • C07F9/65505—Phosphonic acids containing oxirane groups; esters thereof

Definitions

• V ⁇ 9/V ⁇ 2 T cells can also be activated through endogenous metabolites (acting in the micromolar range) such as isopentenyl pyrophosphate or IPP (Espinosa et al., 2001b; Tanaka et al., 1995), which is produced through the conventional mevalonate pathway shared by both microorganisms and mammalian cells. Production of IPP in the latter cells can be up-regulated in situations of cell stress and transformation. In particular a recent study has reported a correlation between the endogenous production levels of IPP in tumor cells and their susceptibility to V ⁇ 9/V ⁇ 2 T cell- mediated lysis (Gober et al., 2003).
• R 3 , R , and R 5 are a hydrogen or (C ⁇ -C 3 )alkyl group
• W is -CH- or -N-
• Re is an (C 2 -C 3 )acyl, an aldehyde, an (C ⁇ -C 3 )alcohol, or an (C 2 -C 3 )ester
• Cat+ represents one (or several, identical or different) organic or mineral cation(s) (including the proton)
• B is O or NH
• m is an integer from 1 to 3
• Y is O " Cat+, a nucleoside, or a radical -A-R, wherein A is O, NH, CHF, CF 2 or CH 2 , and R is selected from the group consisting of 1), 2) or 3).
• X is an halogen (preferably selected from I, Br and Cl)
• B is O or NH
• m is an integer from 1 to 3
• RI is a methyl or ethyl group
• Cat+ represents one (or several, identical or different) organic or mineral cation(s) (including the proton)
• n is an integer from 2 to 20
• Y is O " Cat+, a nucleoside, or a radical -A-R, wherein A is O, NH, CHF, CF 2 or CH 2 andR is selected from the group consisting of 1), 2) or 3).
• the ⁇ T cell activator is a compound of formula (fll)
• the ⁇ T cell activator is a compound of formula (V)
• RI is a methyl or ethyl group
• Cat+ represents one (or several, identical or different) organic or mineral cation(s) (including the proton)
• B is O or NH
• m is an integer from 1 to 3
• n is an integer from 2 to 20
• Y is O " Cat+, a nucleoside, or a radical -A-R, wherein A is O, NH, CHF, CF 2 or CH 2 , and R is selected from the group consisting of 1), 2) or 3).
• the present invention also provides pharmaceutical composition comprising a ⁇ T cell activator according to any one of the embodiments described herein. Also provided are methods of modulating, preferably activating, a ⁇ T cell, the method comprising bringing a ⁇ T cell into contact with a ⁇ T cell activating compound described herein.
• compounds of the invention may be used to activate ⁇ T cell in vitro or in vivo.
• Activated ⁇ T cell in vitro may be used in any suitable method following activation, including in therapy or prevention of disease.
• activated ⁇ T cells are administered to a mammal, preferably a human.
• the invention encompasses a method of treatment comprising (a) bringing a ⁇ T cell into contact with a ⁇ T cell activating compound described herein and (b) administering ⁇ T cells of step (a) to a subject.
• Methods for preparing ⁇ T cells for such applications are known in the art, for example can be carried out as described US 10/505,252, filed August 19 th and 2004 PCT/FR 03/00585 filed February 21 st 2003, both by by Romagne and Laplace, the disclosures of which are incorporated herein by reference.
• the inventions provides a method for treating or preventing a disease comprising administering to a subject a ⁇ T cell activator described herein in an amount sufficient to ameliorate or prevent said disease.
• a ⁇ T cell activator of the invention for the manufacture of a pharmaceutical composition for regulating ⁇ T cells in a human subject.
• said disease is a tumor or proliferative disorder, an infectious disease, an autoimmune disease or an allergic disease.
• the invention further provides methods for the synthesis of phosphoroamidate compounds.
• the invention provides a method for preparing a diphosphoramidate monoester compound comprising: (a) reacting an alkylhalide R-X in a coupling step with a diethylphosphoramidate or diethylchlorophosphate reagent; (b) reacting the compound prepared in step (a) in a saponification step thereby removing O-ethyl groups; and (c) reacting the compound prepared in step (b) in a phosphorylation step thereby preparing a diphosphoramidate monoester, wherein R is a linear, branched, or cyclic, aromatic or not, saturated or unsaturated,
• C1-C50 hydrocarbon group optionally interrupted by at least one heteroatom
• said hydrocarbon group comprises an alkyl, an alkylenyl, or an alkynyl, preferably an alkyl or an alkylene, which can be substituted by one or several substituents selected from the group consisting of : an alkyl, an alkylenyl, an alkynyl, an epoxyalkyl, an aryl, an heterocycle, an alkoxy, an acyl, an alcohol, a carboxylic group (-COOH), an ester, an amine, an amino group (-NH2), an amide (- CONH2), an imine, a nitrile, an hydroxyl (-OH), a aldehyde group (-CHO), an halogen, an halogenoalkyl, a thiol (-SH), a thioalkyl, a sulfone, a sulfoxide, and a combination thereof, and wherein X
• X is an NH2 group and said R-X compound is reacted in a coupling step with a diethylchlorophosphate compound.
• X is selected from the group consisting of I, Br and Cl.
• the invention provides a method of preparing a (E)-2-(4-azido-2-methylbut-2-en yloxy)tetrahydro-2H-pyran compound, comprising providing a (E)-2-(4-Chloro-2-methylbut-2-en yloxy)tetrahydro-2H-pyran compound and reacting said compound with a sodium azide in a water-pentane biphasic mixture in the presence of phase transfer catalyst. Additional embodiments and details are futher provided herein.
• Figure 1 shows an in vitro dose reponse curve and EC50 values for compound of the invention N- HDMAPP and reference compounds BrHPP and HDMAPP.
• the compound of the invention demonstrates not only 3-4 fold increased potency over the next most potent compounds, but also increased absolute activation of ⁇ T cells as observed by TNF ⁇ release.
• the expression "regulating the activity of ⁇ T cells” designates causing or favoring an increase in the number and/or biological activity of such cells in a subject. Regulating thus includes without limitation modulating (e.g., stimulating) expansion of such cells in a subject and/or, for instance, triggering of cytokine secretion (e.g., TNF ⁇ or IFN ⁇ ).
• ⁇ T cells normally represent between about 1-10% of total circulating lymphocytes in a healthy adult human subject.
• the present invention can be used to significantly increase the ⁇ T cells population in a subject, particularly to reach at least 10%, 12%, 15%, 20%, or 30-90% of total circulating lymphocytes, typically 40-90%, more preferably from 50-90%.
• R is a linear, branched, or cyclic, aromatic or not, saturated or unsaturated, C ⁇ -C 50 hydrocarbon group, optionally interrupted by at least one heteroatom, wherein said hydrocarbon group comprises an alkyl, an alkylenyl, or an alkynyl, preferably an alkyl or an alkylene, which can be substituted by one or several substituents selected from the group consisting of : an alkyl, an alkylenyl, an alkynyl, an epoxyalkyl, an aryl, an heterocycle, an alkoxy, an acyl, an alcohol, a carboxylic group (-COOH), an ester, an amine, an amino group (-NH 2 ), an amide (-CONH 2 ), an imine, a nitrile, an hydroxyl (-OH), a aldehyde group (-CHO), an halogen, an halogenoalkyl, a thiol (-SH), a thioalkyl
• the substituents as defined above are substituted by at least one of the substituents as specified above.
• the substituents are selected from the group consisting of : an (C ⁇ -C 6 )alkyl, an (C 2 - C 6 )alkylenyl, an (C 2 -C 6 )alkynyl, an (C 2 -C 6 )epoxyalkyl, an aryl, an heterocycle, an (C ⁇ -C 6 )alkoxy, an (C 2 -C 6 )acyl, an (C ⁇ -Ce)alcohol, a carboxylic group (-COOH), an (C 2 -Ce)ester, an (C ⁇ -C 6 )amine, an amino group (-NH 2 ), an amide (-CONH 2 ), an (C ⁇ -C 6 )imine, a nitrile, an hydroxyl (-OH), a aldehyde group (-CHO), an halogen, an (C ⁇ -C 6 )halogenoalkyl, a thiol (-SH), a (C
• the substituents are selected from the group consisting of : an (C 3 - C 6 )epoxyalkyl, an (Ci-C 3 )alkoxy, an (C 2 -C 3 )acyl, an (C ⁇ -C 3 )alcohol, an (C 2 -C 3 )ester, an (Q- C 3 )amine, an (C ⁇ -C 3 )imine, an hydroxyl, an halogen, an (C ⁇ -C 3 )halogenoalkyl, and a combination thereof, and a combination thereof.
• R is a (C 3 -C 25 )hydrocarbon group, more preferably a (C 5 -C ⁇ o)hydrocarbon group.
• (C 2 -C 6 )alkynyl includes ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, or 5- hexynyl and the other isomeric forms thereof.
• “Amine” groups correspond to the alkyl groups defined hereinabove bonded to the molecule by an -N- (amine) bond.
• (C ⁇ -C 6 )amine includes methylamine, ethylamine, propylamine, butylamine, pentylamine, hexylamine and the other isomeric forms thereof.
• (C ⁇ -C 3 )amine includes methylamine, ethylamine, and propylamine.
• Thioalkyl groups correspond to the alkyl groups defined hereinabove bonded to the molecule by an -S- (thioether) bond.
• (Ci-C 6 )thioalkyl includes thiomethyl, thioethyl, thiopropyl, thiobutyl, thiopentyl, thiohexyl and the other isomeric forms thereof. includes thiomethyl, thioethyl, and thiopropyl.
• Heteroatom denotes N, S, or O.
• the hydrocarbon group is a cycloalkylenyl such as a cyclopentadiene or a phenyl, or an heterocycle such as a furan, a pyrrole, a thiophene, a thiazole, an imidazole, a triazole, a pyridine, a pyrimidine, a pyrane, or a pyrazine.
• the cycloalkylenyl or the heterocycle is selected from the group consisting of a cyclopentadiene, a pyrrole or an imidazole.
• the cycloalkylenyl or the heterocycle is sustituted by an alcohol.
• said alcohol is a (C ⁇ -C 3 )alcohol.
• n is an integer from 2 to 20, and Ri is a methyl or ethyl group.
• Ri is a methyl or ethyl group.
• n is an integer from 2 to 10, or from 2 to 5.
• RI is a methyl.
• B is O or NH. More preferably, B is O.
• phosphoramidate esters according to the present invention comprise the compounds of formula (IV):
• phosphoramidate esters according to the present invention comprise the compounds offormula (VlT) :
• phosphoramidate esters according to the present invention comprise the compounds of formula (VIH):
• RI is a methyl or ethyl group
• Cat+ represents one (or several, identical or different) organic or mineral cation(s) (including the proton), and n is an integer from 2 to 20.
• RI is a methyl.
• n is 2.
• phosphoramidate esters according to the present invention comprise the compound of formula (LX):
• phosphoramidate esters according to the present invention comprise the compounds of formula (X):
• R 3 , R 4 , and R 5 are a hydrogen or (C ⁇ -C 3 )alkyl group
• W is -CH- or -N-
• Re is an (C 2 -C 3 )acyl, an aldehyde, an (C ⁇ -C 3 )alcohol, or an (C 2 -C 3 )ester
• Cat+ represents one (or several, identical or different) organic or mineral cation(s) (including the proton)
• B is O or NH
• m is an integer from 1 to 3
• Y is O " Cat+, a nucleoside, or a radical -A-R, wherein A is O, NH, CHF, CF 2 or CH 2 , and R is selected from the group of 1), 2) or 3).
• Y is O " Cat+, or a nucleoside. More preferably, Y is O " Cat+. More preferably, R 3 and R 4 are a hydrogen and R 5 is a methyl. More preferably, Re is -CH 2 -OH, -CHO, CO-NH 2 , -NH 2 , or -CO-OCH 3 . Still more preferably, Rg is -CH 2 -OH. More preferably, W is -CH-.
• B is O.
• m is 1 or 2. More preferably, m is 1.
• the double-bond between W and C is in conformation trans (E) or cis (Z). More preferably, the double-bond between W and C is in conformation trans (E).
• phosphoramidate esters according to the present invention comprise the compounds of formula (XI) :
• phosphoramidate esters according to the present invention comprise the compound of formula (XD):
• the synthetic ⁇ T lymphocyte activating compound is selected from the group consisting of N-HDMAPP, N-Epox-PP, and N-BrHPP, more preferably N- HDMAPP and N-BrHPP, still more preferably N-HDMAPP.
• Phosphoramidate esters according to the present invention can be for example prepared by the following reactions (Reactions A, A(l), A(2), A(3), B, C or C(l).
• Diphosphoramidate monoesters according to the present invention can also be prepared using the following two reactions (Reactions A(2), A(3)). These synthetic schemes are preferred for larger scale preparations of diphosphoramidate monoesters and involve the formation of a monophosphoramidate monoester intermediate which is of interest for pharmaceutical development since it is considered as a potential metabolite or degradation product.
• This coupling step can be conducted following the procedure reported by Cox et al (2002), the disclosure of which is incorporated herein by reference.
• the saponification step of reaction A(2) involves a two-step procedure leading to a complete removal of O-Ethyl esters functions. This reaction has to be performed preferably under neutral or basic conditions in order to prevent the hydrolysis of the phosphoramidate linkage (P-NH linkage).
• TMSBr trimethylsilyl bromide
• Bu NF tetrabutylammonium fluoride
• Sym-collidine acid scavenger
• reaction A(3) involves the reaction of an alkylamine precursor with commercially available diethylchlorophosphate in the presence of triethylamine (TEA). This reaction can be conducted following the procedure described in Nikolaides et al, (Conversion of Amines to Phosphoesters: decyl diethyl phosphate, Organic Syntheses, CV 9, 194).
• A, B and C reactions may also be performed in non-aqueous solvents with carbodiimide reagents like DCC ( ⁇ , ⁇ '-dicyclohexylcarbodiimide) using organic salts of di- or triphosphate.
• Carbodiimides have been widely employed in the synthesis of ortho- and pyrophosphate esters, nucleotides, cyclic phosphates, oligoribonucleotides, polynucleotides, nucleoside-5'- phosphoroamidates, and mixed anhydrides (Azzi et ah, 1984), the disclosure of which is incorporated herein by reference.
• reaction procedure can be the following: CH 2 CH
• This reaction can also be used for preparing compound comprising an aldehyde derivative as group Y by choosing, instead of Nucl, a derivative including a protected aldehyde function in the form of an acetal or any other group protecting this function.
• PPP represents the triphosphate group
• R- has the above mentioned meaning
• DMF is dimethylformamide
• Nucl is a nucleoside.
• This reaction can be carried out in conditions similar to those described by Knorre et al.(1976), or by Bloom et al., United States Patent No. 5,639,653 (1997), the disclosures of which are incorporated herein by reference, from alcohol and a nucleotide with formula Nucl-O-PPP.
• a cytokine is administered daily for up to about 10 days, preferably for a period of between about 3 and 10 days, or most preferably for about 7 days.
• the administration of the cytokine begins on the same day (e.g. within 24 hours of) as administration of the ⁇ T cell activator.
• the cytokine is administered each day, while in other aspects the cytokine need not be administered on each day.
• a 4-weekly treatment cycle is preferred.
• the first component is administered for about 4 days, a 3-weekly day treatment cycle is preferred.
• the compounds can be used according to any of the methods described in International Patent Application number PCT/TB2003/006375, filed December 2 nd , 2003, the disclosure of which is incorporated herein by reference.
• the invention also relates to a product comprising a ⁇ T cell activator according to the present invention and an interleukin-2 polypeptide, for separate use, for regulating the activity of ⁇ T cells in a mammalian subject.
• the invention concerns a vaccinal composition comprising a ⁇ T cell activator according to the present invention.
• the invention also concerns the use of a ⁇ T cell activator according to the present invention as a vaccine adjuvant.
• Step 4 Coperet et al, Tetrahedron 1996, 52 (35) 11520-11544; Step 5: Sato et al, Chem. Pharm. Bull, 38(8), 2287-2289 (1990); and Step 6: Miyashita et al, J. Org. Chem., 42 (1977) 3772-3774 (deprotection reaction only).
• Tosyl chloride (4.8 g, 25 mmol) and 4-(NN-dimethylamino-) pyridine (3.4 g, 27.5 mmol) were mixed under magnetic stirring with 90 ml of anhydrous dichloromethane in a 250-ml three-necked flask cooled in an ice bath.
• a solution of 3-methyl-3-buten-l-ol (2.2 g, 25 mmol) in about 10 ml of anhydrous dichloromethane was then slowly introduced with a syringe through a septum in the flask, and the ice bath was then removed.
• the reaction was monitored by silica gel TLC (pentane/ethyl acetate, 85:15 (v/v)).
• Disodium pyrophosphate (0.25 mmol - 1 eq.) and 3-methyl-3-en-l -amine (0.25 mmol - 1 eq.) are dissolved in 3 ml of a 1/1 (v/v) deionized water/acetonitrile mixture and introduced into in a glass reaction vessel.
• 12.5 ml of a 0.2 M deionized water/acetonitrile solution of l-ethyl-3-(3 dimethylaminopropyl) carbodiimide hydrochlori.de (2.5 mmol - 10 eq) are added dropwise using a syringe, while stirring at room temperature.
• the compound N-EpoxPP can be prepared starting from the compound N-BrHPP described in Example 3 by treatment with IM ammonium hydroxide solution (epoxidation reaction) followed by a cationic exchange step on DOWEX 50WX8-200 (Na + form) resin.
• the epoxidation reaction with subsequent purification of the crude product can be conducted according to the experimental protocol provided in WO 00/012519 for the preparation of 3, 4-epoxy-3 -methyl- 1-butyl-diphosphate (EpoxPP).
• EpoxPP 4-epoxy-3 -methyl- 1-butyl-diphosphate
• test compound and reference compounds in this case N-HDMAPP, BrHPP and HDMAPP, as further described below.
• 100 ⁇ l of culture supernatant is taken for TNF ⁇ dosage.
• Measurement of the released TNF ⁇ dosage is performed as described by the manufacturer's instruction in the TNF ⁇ enzyme immunoassay kit (ref. 11121, Immunotech - Beckman Coulter).
• OD at 405nm is read, the OD being proportional to the concentration of released TNF ⁇ in the culture supernatant.
• the data are processed with the Excel software to compare concentration of test compound versus concentration of TNF ⁇ and for the calculation of the EC50 for each test compound.
• the bioactivity of the compound N-HDMAPP was assessed using a TNF ⁇ release assay as described above. In vitro activity is shown in Figure 1. Compounds BrHPP and HDMAPP were included for purpose of comparison. The in vitro EC50 was then assessed in this in vitro relative screening test, where prior assays with calibrated cells using a BrHPP-standard composition presented an EC50 of about 15 nM for BrHPP. As will be appreciated, any other suitable assays such as cell amplification may be used in assessing compounds.
• the EC50 for N-HDMAPP was determined to be 0.63 nM while the in vitro EC50 for HDMAPP was 2.1 nM and the in vitro EC50 for BrHPP was 37.7 nM. Since the assay provides a relative result rather than absolute EC50 value, the results indicate that the N-HDMAPP compound has 3-4 times greater potency that the most potent compounds tested so far.
• the antituberculous Mycobacteriumbovis BCG vaccine is an attenuated mycobacterial producer of phosphorylated nonpeptidic antigens for human gamma delta T cells. Infect Immun 63, 4628-4633.
• Escherichia coli produces phosphoantigens activating human gamma delta T cells. J Biol Chem 277, 148-154.
• Vgamma9/Vdelta2 T cell activation induced by bacterial low molecular mass compounds depends on the 1-deoxy-D-xylulose 5-phosphate pathway of isoprenoid biosynthesis.
• CD94/NKG2 inhibitory receptor complex modulates both anti-viral and anti-tumoral responses of polyclonal phosphoantigen-reactive V gamma 9V delta 2 T lymphocytes. J Immunol 159, 6009-6017.
• Valentijn G. A. van der Marel; L. H. Cohen; J. H. van Boom, Synlett 1991, 663-664.

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• 2004
  • 2004-12-02 CA CA002547008A patent/CA2547008A1/en not_active Abandoned
  • 2004-12-02 EP EP04806475A patent/EP1689758A2/en not_active Withdrawn
  • 2004-12-02 AU AU2004295194A patent/AU2004295194A1/en not_active Abandoned
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  • 2004-12-02 WO PCT/IB2004/004311 patent/WO2005054258A2/en not_active Ceased
  • 2004-12-02 CN CNA2004800356576A patent/CN1890252A/zh active Pending
  • 2004-12-02 US US10/581,144 patent/US7767842B2/en not_active Expired - Fee Related
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