New amphetamine derivatives, antibodies against them and pharmaceutical compositions containing them

• US 20030171435A1
• Filed: 01/23/2002
• Published: 09/11/2003
• Est. Priority Date: 01/23/2002
• Status: Abandoned Application

• Current Assignee: DrugAbuse Sciences SAS (elbion NV)
• Original Assignee: DrugAbuse Sciences SAS (elbion NV)
• Inventors: Blanchard, Dominique, Gadjou, Caroline, Koralewski, Frederic, Pouletty, Philippe, Kusmierek, Jacques, Galons, Herve
• Application Number:
  US10/057,791
• Publication Number:
  US 20030171435A1
• Time in Patent Office: Days
• US Class Current: 514/567
• CPC Class Codes: A61P 39/02 Antidotes
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids

Abstract

Hapten-carrier conjugates capable of eliciting anti-hapten antibodies in vivo to amphetamines are disclosed. Methods of preparing the hapten-carrier conjugates and therapeutic compositions are also disclosed. A therapeutic composition containing the hapten-carrier conjugate is useful in the treatment of addiction to amphetamines. Passive immunization using antibodies raised against conjugates of the instant invention also is disclosed. The therapeutic composition is suitable for co-therapy with other conventional drugs for treatment of amphetamine abuse.

Claims

1. An (S)-enantiomer compound having the formula (I):

2. A compound having formula (II)

3. The compound according to claim 2, having formula (VI):

4. The compound according to claim 2, having formula (VII):

9. A method for obtaining a compound according to claim 4, said method comprising the steps of:

(a) alkylating a methamphetamine derivative having formula (IX) as shown in FIG. 1 by addition of Br(CH₂)_nCOOBn, wherein n is an integer between 1 and 6, whereby an addition product of formula (X) as shown in FIG. 1 is obtained ; and

(b) hydrogenating said addition product of formula (X), whereby a compound according to claim 4 is obtained.

42. A compound according to any one of claims 2 to 8, wherein said compound is the S-enantiomer.

5. A compound having formula (IIIa):

6. The compound according to claim 5 having formula (IV):

10. A method for obtaining a compound according to claim 5, said method comprising the steps of:

(a) condensing a a compound of formula (XI) as shown in FIG. 2, with CH₃CH₂—

NO₂, whereby a nitrostyrene derivative of formula (XII) as shown in FIG. 2 is obtained;

(b) reducing said nitrostyrene derivative (XII), whereby an amphetamine derivative of formula (XIII) as shown in FIG. 2 is obtained;

(c) alkylating the primary amine of said amphetamine derivative whereby a compound of formula (XIV) as shown in FIG. 2 is obtained;

(d) acylating the amine group on said amphetamine derivative (XIII) or said compound of formula (XIV), whereby a protected derivative of a compound having formula (XV) as shown in FIG. 2 is obtained;

(e) hydrogenating said protected derivative (XV), whereby a compound having a formula (XVI) as shown in FIG. 2 is obtained;

(f) alkylating the phenol groups of said compound of formula (XVI) with benzylbromoalkylcarboxylate, whereby a benzylester compound of formula (XVII) as shown in FIG. 2 is obtained;

(g) hydrogenating said benzylester compound of formula (XVII) whereby a carboxylic derivative of formula (XVIII) as shown in FIG. 2 is obtained;

(h) deprotecting the amino group of said carboxylic acid derviative (XVIII); and

(i) separating the enantiomers so obtained, whereby compounds according to claim 5 are obtained.

11. The method according to claim 10 steps (a) through (g) further comprising the steps of:

(h) acylating (R)H—

cys(Otrt)—

NH2 with said carboxylic derivative (XVIII);

(i) separating the resulting diastereoisomer mixture whereby an S-isomer having formula (XIX) as shown in FIG. 2 is obtained; and

j) removing acid-sensitive protecting groups from said S-isomer having formula (XIX), whereby a compound according to claim 7 is obtained.

12. The method according to claim 10 or claim 11, wherein said alkylating step (c) comprises:

contacting said compound of formula (XIII) with trifluoroacetic anhydride;

alkylating the resulting trifluoroacetaraide with an alkyl halide; and

removing the trifluoroacetyl group in basic medium.

13. The method according to claim 10 or claim 11, wherein said alkylating step (c) comprises:

contacting said compound of formula (XIII) with an acid anhydride to obtain an amide; and

reducing said amide with lithium aluminium hydride.

14. A method for obtaining a compound according to claim 5, said method comprising the steps of:

(a) reducing a tyrosine ester derivative of a compound of formula (XXI) as shown in FIG. 3, whereby an alcohol derivative of formula (XXII) as shown in FIG. 3 is obtained;

(b) alkylating said alcohol derivative (XXIII) whereby a compound of formula (XXIV) as shown in FIG. 3 is obtained;

(c) hydrogenating said compound of formula (XXIV) whereby a phenol derivative of formula (XXV) as shown in FIG. 3 is obtained;

(d) contacting said phenyl derivative (XXV) with benzylbromoalkylcarboxylate whereby an ester derivative of formula (XXVI) as shown in FIG. 3 is obtained; and

(e) hydrogenating said ester derivative (XXVI) whereby a compound according to claim 5 is obtained.

7. A compound having formula (IIIb)

8. The compound according to claim 7 having formula (V):

15. An amphetamine-related immunogen comprising:

a compound having formula (I) covalently conjugated to a carrier.

19. An amphetamine-related immunogen according to any one of claims 15 to 18, wherein said carrier is a protein selected from the group consisting of tetanus toxin, bovine serum albumin, ovalbumin, KLH, or a peptidic fragment of any of the proteins in said group.

21. A pharmaceutical composition comprising:

at least one amphetamine-related immunogen according to claim 15.

16. An amphetamine-related immunogen I1 comprising:

a compound having formula (II) covalently conjugated to a carrier,

 

wherein p is an integer between 1 and 6;

wherein R₁ and R₃ are selected from the group consisting of hydrogen, and C₁-C₃ alkyl;

R₆ and R₇ are the same or different and are selected from the group consisting of hydrogen, a halogen, —

OR₉ and —

SR₉, wherein R₉ is hydrogen or a C1-C₃ alkyl; and

R₅ is selected from the group consisting of hydrogen, a polymethylene chain;

—

(CH)_m—

R₁₀ and an oxy-polymethylene chain;

—

O—

(CH₂)_m—

R₁₀, wherein —

R₁₀ is selected from the group consisting of carboxyl, thiol, —

CONHR₁₃SH, and —

CONHCHR₁₁SH, wherein R₁₃ is selected from the group consisting of —

CH(COOH)CH₂—

and —

(CH₂)_m— and

wherein m is an integer between 1 and 4, with the proviso that when R₁ is hydrogen and R₂ is a methyl or when R₁ is a methyl and R₂ is hydrogen, then R₅ is not a polymethylene chain;

—

(CH₂)_m—

COOH.

17. An amphetamine-related immunogen 12 comprising:

a compound having formula (IIIa) covalently conjugated to a carrier,

 

wherein R₁ and R₃ are selected from the group consisting of hydrogen, and C₁-C₃ alkyl;

R₂ is selected from the group consisting of hydrogen, a C₁-C₃ alkyl and a polymethylene chain;

(CH₂)_n—

COOH, wherein n is an integer between 1 and 6;

R₄, R₆ and R₇ are the same or different and are selected from the group consisting of hydrogen, a halogen, —

OR₉ and —

SR₉, wherein R₉ is hydrogen or a C₁-C₃ alkyl; and

,

R₁₁ is selected from the group consisting of a polymethylene chain;

—

(CH₂)_m— and

an oxy-polymethylene chain;

—

O(CH₂)_m—

, wherein m is an integer between 1 and 4, with the proviso that when R₁ is a methyl, then R ₁ is not a polymethylene chain;

—

(CH₂)_m—

.

18. An amphetamine-related immunogen I3 comprising:

a compound having formula (IIIb) covalently conjugated to a carrier,

 

wherein R₁ and R₃ are selected from the group consisting of hydrogen, and a C₁-C₃ alkyl;

R₂ is selected from the group consisting of hydrogen, a C₁-C₃ alkyl and a polymethylene chain;

(CH₂)_n—

COOH, wherein n is an integer between 1 and 6;

R₄, R₆ and R₇ are the same or different and are selected from the group consisting of hydrogen, a halogen, —

OR₉ and —

SR₉, wherein R₉ is hydrogen or a C₁-C₃ alkyl; and

R₁₂ is selected from the group consisting of a polymethylene chain;

(CH₂)_mCONHR₁₃ and an oxy-polymethylene chain;

—

O(CH₂)_mCONHCH—

R₁₃;

wherein R₁₃ is selected from the group consisting of —

CH(COOH)CH₂— and

—

(CH₂)_m—

, wherein m is an integer between 1 and 4, with the proviso that when R₁ is hydrogen and R₂ is a methyl or when R₁ is a methyl and R₂ is hydrogen, then R₅ is not a polymethylene chain;

—

CH₂)_m.

22. A vaccine comprising at least one amphetamine-related immunogens according to claim 18 or claim 20.

41. A method of prophylactic treatment of abuse of at least one amphetamine in a patient in need thereof, said method comprising:

immunizing said patient with a vaccine according to claim 22.

20. The amphetamine-related immunogen TT-Met1 having the formula

23. Antibodies or fragments or chains of immunoglobulins having affinity for one or more amphetamine or derivative of amphetamine.

24. The antibodies according to claim 23, wherein said antibodies are monoclonal antibodies.

25. The antibodies according to claim 23 or claim 24, wherein said antibodies are neutralizing antibodies.

26. The antibodies according to claim 24, wherein said antibodies are capable of binding to at least two different amphetamines or amphetamines derivatives.

27. The antibodies according to claim 26, wherein said at least two different amphetamines or amphetmine derivatives comprise methamphetamine and ecstasy.

30. A composition comprising:

a cell line producing a monoclonal antibody according to claim 24.

31. The composition according to claim 30, wherein said antibodies are murine antibodies.

36. Monoclonal antibodies according to claim 24, labelled with an agent capable of providing a detectable signal.

38. A pharmaceutical composition comprising:

at least one antibody according to claim 23, capable of binding to one or more amphetamine derivatives.

40. The method according to claim 38, wherein said symptoms are a result of ingesting two or more amphetamine derivatives and said at least one antibody is capable of binding to at least two of said two or more amphetamine derivatives.

28. A monoclonal antibody produced by hybridoma DASM243-6H5D1C4.

29. A monoclonal antibody produced by hybridoma DASM243-3A10 A6A2.

32. The murine hybridoma DASM243-645D1 C4 having accession number CNCM I-2750.

33. A cell line which produces monoclonal antibodies having the same antigenic specificity as that of monoclonal antibodies expressed by cell line DASM243-645D1C4.

34. Monoclonal antibodies derived from a hybridoma according to claim 33.

35. An imnmunoglobulin fragment or chain derived from a monoclonal antibody having the binding specificity of a monoclonal antibody according to claim 34.

37. An imnmunoglobulin fragment or chain according to claim 35, labelled with an agent capable of providing a detectable signal.

39. A method for ameliorating symptoms of abuse of at least one amphetamine in a patient in need thereof said method comprising:

providing said patient with a pharmaceutical composition according to claim 37, in an amount sufficient to ameliorate said symptoms.

43. Isolated nucleic acid encoding the heavy and light chains of monoclonal antibody DASM243-645D1C4 or DASM243-3A10A6A2.

46. The isolated nucleic acid according to claim 43, wherein said nucleic acid is DNA.

47. The isolated nucleic acid according to claim 43, wherein said nucleic acid molecule is cDNA.

48. An expression vector comprising:

nucleic acid according to claim 43 operably linked to transcriptional and translational regulatory regions.

49. A host cell comprising:

an expression vector according to claim 48.

50. The host cell according to claim 49, wherein said cell is a eukaryotic cell.

44. Isolated nucleic acid encoding a polypeptide derived from monoclonal antibody DASM243-645D1C4 or DASM243-3A10A6A2.

45. The isolated nucleic acid according to claim 44, wherein said polypeptide is a light chain protein.

51. A process for producing an immunoglobulin specific for at least two amphetamines or amphetamine derivatives or an immunologically functional immunoglobulin fragment thereof comprising at least the variable domains of the heavy and light chains of said immunoglobulin, said method comprising:

growing a host cell comprising expression vectors comprising nucleic acid encoding respectively the heavy and light chains of monoclonal antibody DASM243-645D1C4 or DASM243-3A1 or functional fragments thereof, so that said nucleic acid is expressed, whereby said immunoglobulin is obtained.