IMPROVED METHOD FOR THE PRODUCTION OF LYSERGIC ACID DIETHYLAMIDE (LSD) AND NOVEL DERIVATIVES THEREOF

Abstract

The present invention provides an improved method for the production of lysergic acid diethylamide (LSD) for GMP purposes. Furthermore, the present invention provides novel LSD derivatives of formula I as well as their synthesis and purification. Due to the affinity of the presented substances for the 5-HT.sub.2A receptor, the invention may find application in numerous forms of therapy, such as against depression or drug addiction.

Claims

1. A method for the production of lysergic acid diethylamide (LSD), comprising the steps of: a. preparing a suspension of lysergic acid hydrate in ethyl acetate; b. addition of diethylamine under protective gas atmosphere; c. addition of propane-phosphonic acid anhydride solution (T3P) in ethyl acetate; d. stirring of the mixture under protective gas atmosphere for at least 4 hours; e. stopping the reaction by dilution with ethyl acetate; f. extraction with water; g. drying of the organic phase over a desiccant at 20-60° C. and under vacuum; h. obtaining a crude product containing lysergic acid diethylamide (LSD).

2. A method for the production of a lysergic acid diethylamide (LSD) derivative of the following formula II ##STR00039## wherein: R.sub.N is selected from —NH—(C.sub.1-5 alkyl), —N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), —NH—(C.sub.1-5 haloalkyl), —N(C.sub.1-5 alkyl)(C.sub.1-5 haloalkyl), —N(C.sub.1-5 haloalkyl)(C.sub.1-5 haloalkyl), —NH—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl), —N(C.sub.1-5 alkyl)[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)], —N[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)]-(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl), —N(C.sub.1-5 haloalkyl)[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)], —N(C.sub.3-7 cycloalkyl)(C.sub.3-7 cycloalkyl), an N-containing polycyclic heterocyclyl, 1,3-oxazolidin-3-yl, 3-methylpyrrolidin-1-yl, and an N-containing monocyclic heterocyclyl which is substituted with one or more halogens, wherein any alkyl groups and/or any alkylene groups comprised in said —NH—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl), in said —N(C.sub.1-5 alkyl)[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)] or in said —N[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)]-(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl) are each optionally substituted with one or more halogens, wherein said N-containing polycyclic heterocyclyl or said N-containing monocyclic heterocyclyl comprises at least one nitrogen ring atom and is attached to the remainder of the compound via said nitrogen ring atom, wherein said N-containing polycyclic heterocyclyl is not indolin-1-yl, 1,2,3,4-tetrahydroquinolin-1-yl or 3-azabicyclo[3.2.2]nonan-3-yl, wherein said N-containing polycyclic heterocyclyl, said 1,3-oxazolidin-3-yl, said N-containing monocyclic heterocyclyl, and the cycloalkyl groups comprised in said —N(C.sub.3-7 cycloalkyl)(C.sub.3-7 cycloalkyl) are each optionally substituted with one or more groups R.sub.4, and further wherein R.sub.N is not —N(CH.sub.2CH.sub.3)—CH.sub.2CH.sub.3; and each R.sub.4 is independently selected from C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, —(C.sub.0-3 alkylene)-OH, —(C.sub.0-3 alkylene)-O(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-O(C.sub.1-5 alkylene)-OH, —(C.sub.0-3 alkylene)-O(C.sub.1-5 alkylene)-O(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-SH, —(C.sub.0-3 alkylene)-S(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH.sub.2, —(C.sub.0-3 alkylene)-NH(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH—OH, —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-OH, —(C.sub.0-3 alkylene)-NH—O(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-O(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-halogen, —(C.sub.0-3 alkylene)-(C.sub.1-5 haloalkyl), —(C.sub.0-3 alkylene)-O—(C.sub.1-5 haloalkyl), —(C.sub.0-3 alkylene)-CN, —(C.sub.0-3 alkylene)-NO.sub.2, —(C.sub.0-3 alkylene)-CHO, —(C.sub.0-3 alkylene)-CO—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-COOH, —(C.sub.0-3 alkylene)-CO—O—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-O—CO—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-CO—NH.sub.2, —(C.sub.0-3 alkylene)-CO—NH(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-CO—N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH—CO—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-CO—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH—CO—O—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-CO—O—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-O—CO—NH—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-O—CO—N(C.sub.1-5 alkyl)-(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-SO.sub.2—NH.sub.2, —(C.sub.0-3 alkylene)-SO.sub.2—NH(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-SO.sub.2—N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH—SO.sub.2—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-SO.sub.2—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-SO.sub.2—(C.sub.1-5 alkyl), and —(C.sub.0-3 alkylene)-SO—(C.sub.1-5 alkyl); wherein the method comprises the steps of:  a. preparing a suspension of lysergic acid hydrate in ethyl acetate;  b. addition of an amine compound of the formula R.sub.N—H, wherein R.sub.N has the same meaning as in formula II, under protective gas atmosphere;  c. addition of propane-phosphonic acid anhydride solution (T3P) in ethyl acetate;  d. stirring of the mixture under protective gas atmosphere for at least 4 hours;  e. stopping the reaction by dilution with ethyl acetate;  f. extraction with water;  g. drying of the organic phase over a desiccant at 20-60° C. and under vacuum;  h. obtaining a crude product containing the LSD derivative of formula II.

3. The method according to claim 2, wherein the steps a. to f. are carried out at 25° C.

4. The method according to claim 3, wherein 10 equivalents of diethylamine are used.

5. The method according to claim 2, wherein the crude product is subsequently subjected to a method for isomer optimization, comprising the steps of: a. dissolving the crude product in ethanol, b. addition of sodium methoxide, c. stirring for at least 2 hours, d. dilution with water, e. distillation of the solvent, f. redilution of the residue with water, g. extraction with ethyl acetate, h. drying of the organic phase over a desiccant at 40-60° C. and under vacuum, i. obtaining the isomer-optimized intermediate product.

6. The method according to claim 5, wherein the isomer-optimized intermediate product is subsequently subjected to a column purification process using a toluene/ethanol mixture.

7. (canceled)

8. A lysergic acid diethylamide (LSD) derivative produced by the method according to claim 2.

9. A compound which is a lysergic acid diethylamide (LSD) derivative according to the general formula I: ##STR00040## or a pharmaceutically acceptable salt thereof, wherein: R.sub.1 is selected from —NH—(C.sub.1-5 haloalkyl), —N(C.sub.1-5 alkyl)(C.sub.1-5 haloalkyl), —N(C.sub.1-5 haloalkyl)(C.sub.1-5 haloalkyl), —NH—CH.sub.2—O—(C.sub.1-5 alkyl), —NH—(CH.sub.2).sub.3-5—O—(C.sub.1-5 alkyl), —N(C.sub.1-5 alkyl)[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)], —N[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)]-(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl), —N(C.sub.1-5 haloalkyl)[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)], —N(C.sub.3-7 cycloalkyl)(C.sub.3-7 cycloalkyl), an N-containing polycyclic heterocyclyl, 1,3-oxazolidin-3-yl, 3-methylpyrrolidin-1-yl, and an N-containing monocyclic heterocyclyl which is substituted with one or more halogens, wherein said —NH—(C.sub.1-5 haloalkyl) is not —NH—CH.sub.2CH.sub.2—Cl or —NH—CH(—CH.sub.2CH.sub.3)—CH.sub.2—Cl, wherein any alkyl groups and/or any alkylene groups comprised in said —NH—CH.sub.2—O—(C.sub.1-5 alkyl), in said —NH—(CH.sub.2).sub.3-5—O—(C.sub.1-5 alkyl), in said —N(C.sub.1-5 alkyl)[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)] or in said —N[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)]-(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl) are each optionally substituted with one or more halogens, wherein said N-containing polycyclic heterocyclyl or said N-containing monocyclic heterocyclyl comprises at least one nitrogen ring atom and is attached to the remainder of the compound of formula I via said nitrogen ring atom, wherein said N-containing polycyclic heterocyclyl is not indolin-1-yl, 1,2,3,4-tetrahydroquinolin-1-yl or 3-azabicyclo[3.2.2]nonan-3-yl, and further wherein said N-containing polycyclic heterocyclyl, said 1,3-oxazolidin-3-yl, said N-containing monocyclic heterocyclyl, and the cycloalkyl groups comprised in said —N(C.sub.3-7 cycloalkyl)(C.sub.3-7 cycloalkyl) are each optionally substituted with one or more groups R.sub.4, and R.sub.2 is selected from C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, and C.sub.1-5 haloalkyl; or alternatively R.sub.1 is —NH—(C.sub.1-5 alkyl) or —N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), and R.sub.2 is C.sub.1-5 haloalkyl; R.sub.3 is selected from hydrogen, C.sub.1-5 alkyl, —CO—(C.sub.1-5 alkyl), —CO—(C.sub.3-6 cycloalkyl), and an amino acid, wherein said amino acid is attached via a —CO— group formed from a carboxylic acid group of the amino acid, and further wherein said C.sub.1-5 alkyl, the alkyl group comprised in said —CO—(C.sub.1-5 alkyl), the cycloalkyl group comprised in said —CO—(C.sub.3-6 cycloalkyl) and any alkyl group comprised in said amino acid are each optionally substituted with one or more halogens; and each R.sub.4 is independently selected from C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, —(C.sub.0-3 alkylene)-OH, —(C.sub.0-3 alkylene)-O(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-O(C.sub.1-5 alkylene)-OH, —(C.sub.0-3 alkylene)-O(C.sub.1-5 alkylene)-O(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-SH, —(C.sub.0-3 alkylene)-S(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH.sub.2, —(C.sub.0-3 alkylene)-NH(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH—OH, —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-OH, —(C.sub.0-3 alkylene)-NH—O(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-O(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-halogen, —(C.sub.0-3 alkylene)-(C.sub.1-5 haloalkyl), —(C.sub.0-3 alkylene)-O—(C.sub.1-5 haloalkyl), —(C.sub.0-3 alkylene)-CN, —(C.sub.0-3 alkylene)-NO.sub.2, —(C.sub.0-3 alkylene)-CHO, —(C.sub.0-3 alkylene)-CO—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-COOH, —(C.sub.0-3 alkylene)-CO—O—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-O—CO—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-CO—NH.sub.2, —(C.sub.0-3 alkylene)-CO—NH(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-CO—N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH—CO—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-CO—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH—CO—O—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-CO—O—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-O—CO—NH—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-O—CO—N(C.sub.1-5 alkyl)-(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-SO.sub.2—NH.sub.2, —(C.sub.0-3 alkylene)-SO.sub.2—NH(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-SO.sub.2—N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-NH—SO.sub.2—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-N(C.sub.1-5 alkyl)-SO.sub.2—(C.sub.1-5 alkyl), —(C.sub.0-3 alkylene)-SO.sub.2—(C.sub.1-5 alkyl), and —(C.sub.0-3 alkylene)-SO—(C.sub.1-5 alkyl).

10. The compound according to claim 9, wherein: R.sub.1 is selected from —NH—(C.sub.1-5 haloalkyl), —N(C.sub.1-5 alkyl)(C.sub.1-5 haloalkyl), —N(C.sub.1-5 haloalkyl)(C.sub.1-5 haloalkyl), —N(C.sub.1-5 haloalkyl)[—(C.sub.1-5 alkylene)-O—(C.sub.1-5 alkyl)], —N(—CH.sub.3)—CH.sub.2CH.sub.2—O—CH.sub.3, —N(cyclopropyl)(cyclopropyl), an N-containing polycyclic heterocycloalkyl, 1,3-oxazolidin-3-yl, 3-methylpyrrolidin-1-yl, and an N-containing monocyclic heterocycloalkyl which is substituted with one or more halogens, wherein said —NH—(C.sub.1-5 haloalkyl) is not —NH—CH.sub.2CH.sub.2—Cl or —NH—CH(—CH.sub.2CH.sub.3)—CH.sub.2—Cl, wherein said N-containing polycyclic heterocycloalkyl or said N-containing monocyclic heterocycloalkyl comprises at least one nitrogen ring atom and is attached to the remainder of the compound of formula I via said nitrogen ring atom, wherein said N-containing polycyclic heterocycloalkyl is not 3-azabicyclo[3.2.2]nonan-3-yl, and further wherein said N-containing polycyclic heterocycloalkyl, said 1,3-oxazolidin-3-yl, and said N-containing monocyclic heterocycloalkyl are each optionally substituted with one or more groups R.sub.4; and R.sub.2 is selected from C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, and C.sub.1-5 haloalkyl; preferably wherein R.sub.2 is methyl or —CH.sub.2CH.sub.2F.

11. The compound according to claim 9, wherein R.sub.1 is —NH—(C.sub.1-5 alkyl) or —N(C.sub.1-5 alkyl)(C.sub.1-5 alkyl), and wherein R.sub.2 is C.sub.1-5 haloalkyl; preferably wherein R.sub.2 is —CH.sub.2CH.sub.2F.

12. The compound according to claim 9, wherein R.sub.3 is hydrogen, —CO—(C.sub.1-5 alkyl), or —CO—(C.sub.3-6 cycloalkyl).

13. The compound according to claim 9, wherein: R.sub.1 is selected from ##STR00041## R.sub.2 is selected from —CH.sub.3 and —CH.sub.2CH.sub.2F; and R.sub.3 is selected from —H, —COCH.sub.3 and —COCH.sub.2CH.sub.3; or alternatively R.sub.1 is ##STR00042## R.sub.2 is —CH.sub.2CH.sub.2F, and R.sub.3 is selected from —H, —COCH.sub.3 and —COCH.sub.2CH.sub.3.

14. The compound according to claim 9, wherein said compound has the following absolute configuration: ##STR00043##

15. The compound according to claim 9, wherein said compound is selected from any one of the following compounds: ##STR00044## ##STR00045## ##STR00046## or a pharmaceutically acceptable salt thereof.

16. A pharmaceutical composition comprising the lysergic acid diethylamide (LSD) derivative according to claim 8, and optionally one or more pharmaceutically acceptable excipients.

17. (canceled)

18. (canceled)

19. (canceled)

20. (canceled)

21. (canceled)

22. (canceled)

23. (canceled)

24. (canceled)

25. (canceled)

26. A method of treating a serotonin 5-HT.sub.2A receptor associated disease/disorder in a subject in need thereof, the method comprising administering a therapeutically effective amount of the lysergic acid diethylamide (LSD) derivative according to claim 8 to said subject.

27. The method according to claim 26, wherein said disease/disorder is an anxiety disorder, attention deficit hyperactivity disorder (ADHD), depression, cluster headache, a condition associated with cancer, diminished drive, burn-out, bore-out, migraine, Parkinson's disease, dementia, pulmonary hypertension, schizophrenia, an eating disorder, nausea, or vomiting.

28. (canceled)

29. The method according to claim 27, wherein said disease/disorder is an anxiety disorder, attention deficit hyperactivity disorder (ADHD), depression, cluster headache, a condition associated with cancer, diminished drive, burn-out, bore-out, migraine, Parkinson's disease, dementia, pulmonary hypertension, schizophrenia, an eating disorder, nausea, or vomiting.

30. (canceled)

Description

BRIEF DESCRIPTION OF THE FIGURES

[0195] FIG. 1: A sample of the reaction mixture of Example 1 shows the following components in LC/MS (liquid chromatography-mass spectrometry) at a wavelength of 312 nm: 0.85% lysergic acid; 69.4% LSD; 29.7% iso-LSD.

[0196] FIG. 2: TLC (thin layer chromatography) evaluation of fractions 1-10 of the reaction product following column chromatographic purification (see Example 3).

[0197] FIG. 3: TLC evaluation of fractions 10-13 of the reaction product following column chromatographic purification (see Example 3).

[0198] FIG. 4: Production of lysergic acid-mono-(2,2,2-trifluoroethyl)amide (see Example 5), TLC shows conversion after 24 h.

[0199] FIG. 5: TLC evaluation of the production of lysergic acid-mono-(2,2,2-trifluoroethyl)amide following purification. According to LC-MS, fractions 1-7 contain 99% pure product.

[0200] The invention will now be described by reference to the following examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention.

EXAMPLES

Example 1: Method of Production of Lysergic Acid Diethylamide (LSD)

[0201] Lysergic acid hydrate (10 mmol/2.86 g) is suspended in ethyl acetate (200 ml) at 25° C.

[0202] Diethylamine (100 mmol/10.4 ml) is added and aerated with argon. This results in a whitish-gray suspension.

[0203] A 50% (wt %) solution of propane-phosphonic acid anhydride (T3P) in ethyl acetate (30 mmol/19.08 g) is added dropwise through the septum during 70 min.

[0204] During the addition, the initial suspension already starts to turn clear at less than 2 ml propane-phosphonic acid anhydride solution, and a few minutes after the addition, a clear solution is obtained. Thereby, a slightly warm shading can be observed.

[0205] The immediate dissolution of the otherwise not easily dissoluble lysergic acid may be explained by the formation of intermediary lysergic acid ethyl ester, which is subsequently detectable in the crude product as traces. Stirring is continued for further 2.5 h under argon at 26° C.

[0206] A sample of the reaction mixture shows the following components in LC/MS at a wavelength of 312 nm: 0.85% lysergic acid; 69.4% LSD; 29.7% iso-LSD (see also FIG. 1).

[0207] The reaction is stirred for further 3 h at 26° C. under argon, followed by dilution with ethyl acetate (200 ml) to stop the reaction. The dark yellow slightly colloidal reaction solution is extracted with 150 ml water. The aqueous phase has a pH of 9.

[0208] The organic phase is dried over little MgSO.sub.4 and concentrated at 45° C. at up to 40 mbar.

[0209] This yields 4.1 g of a yellow-brown oil (crude product).

[0210] HPLC at 312 nm shows 99.9% of product (both isomers) and 0.1% of ethyl ester.

Example 2: Isomer Optimization

[0211] The complete amount is dissolved in absolute ethanol (60 ml). Sodium methoxide (520 mg) (0.95 eq) is added and stirred for 3 h at 50° C.

[0212] After two hours, sampling for HPLC yielded 82% of LSD.

[0213] After 3.5 hours, sampling for HPLC yielded 84%.

[0214] After 4 hours, the reaction was stopped. At this time, the isomer ratio according to HPLC at 312 nm has changed to 84%:16% of LSD to iso-LSD.

[0215] The product is diluted with water (100 ml) and rotated in (i.e. the water is distilled off on a rotary evaporator) at pH 11-11.5. The aqueous residue is again diluted with water (100 ml) and extracted four times with 150 ml ethyl acetate.

[0216] The product is dried over magnesium sulfate and then concentrated.

[0217] This yields 3.8 g of a brown oil.

[0218] Other concentrations of NaOMe and different times were tried as well.

[0219] The equilibrium of >80% LSD to iso-LSD had adjusted after 2-3 h. Even after 12 h, this ratio had not significantly improved. However, signs of decomposition could already be seen after 12-24 hours.

Example 3: Column Chromatographic Purification

[0220] Isomer-optimized crude material is column treated over 300 g silica using the eluent mixture toluene/ethanol at 95:5.

[0221] Crude product is dissolved in toluene/ethanol at 70:30 (15 ml) and applied.

[0222] The column is started with toluene/ethanol at 95:5. Following one liter of this eluent it is changed to toluene/ethanol at 90:10. One of the impurities runs first as a green band (365 nm). The product runs directly behind it as a bright violet region (365 nm) (see FIGS. 2 and 3).

[0223] The product fractions are concentrated, on a rotary evaporator. Thereby, 2.9 g of a bright foam is obtained. This corresponds to 89% of the theoretical yield.

[0224] HPLC at 312 nm confirms a purity of more than 99%. No iso-LSD at all is present anymore.

Example 4: Production of LSD Derivatives

[0225] The novel LSD derivatives according to the present invention can be produced using the method of production of the invention in high purity and yield.

[0226] For this purpose, a person skilled in the art merely has to select the respective amine compound (in part as a hydrochloride) accordingly. Instead of diethylamine the person skilled in the art has to use 10 eq amine HCl+10 eq DiPEA. The amine to be reacted is produced in situ in this way.

[0227] The purity to be expected is more than 95% and the yield is at least 50%.

Example 5: Alternative Production of Lysergic Acid-Mono-(2,2,2-Trifluoroethyl)Amide

[0228] In order to demonstrate the producibility of the LSD derivatives with conventional methods as well, a few derivatives were synthesized by using conventional methods of production. The compound is producible, but the yields are significantly lower than by using the method of the invention.

[0229] Lysergic acid hydrate (11 mmol; 2.94 g) is suspended in dichloromethane (200 ml) at 26° C. and is aerated with argon. Diisopropylethylamine (11 mmol; 1.9 ml) is added dropwise through the septum.

[0230] Then, carbodiimidazole (17 mmol; 2.75 mg) is colloidally dissolved in dichloromethane (60 ml) and added dropwise through the septum.

[0231] This is stirred for 20 min at 26° C. Then, 2,2,2-trifluoroethylamine base (22 mmol; 1.73 ml) is added dropwise through the septum and stirred for 24 h at 26° C. TLC shows a conversion after 24 h (see FIG. 4).

[0232] On the next day, the reaction is stopped with 2% ammonia solution (30 ml) and the reaction mixture is diluted with dichloromethane (100 ml). Then, the organic phase is separated and washed with water (100 ml) and saturated saline (100 ml). Thereafter, it is dried over magnesium sulfate and concentrated on the rotary evaporator.

[0233] Thereby, 2.5 g of a brown oil is obtained.

[0234] The purification is done by means of a 150 g silica column using toluene/ethanol at 95:5 as an eluent.

[0235] The crude product is dissolved in toluene/dichloromethane at 1:1 (10 ml) and applied. The column is started with pure toluene (300 ml). Thereafter, the eluent is changed to toluene/ethanol at 95:5. According to LC-MS, fractions 1-7 contain 99% pure product (see FIG. 5).

[0236] Without previous isomer optimization, about 0.7 g of a brown oil is obtained. This corresponds to a yield of 18%.

Example 6: Alternative Production of 2-oxa-6-azaspiro[3.3]heptyl-lysergic acid amide

[0237] In order to demonstrate the producibility of the LSD derivatives with conventional methods as well, a few derivatives were synthesized by using conventional methods of production. The compound is producible, but the yields are significantly lower than by using the method of the invention.

[0238] Lysergic acid hydrate (3 mmol; 858 mg) is suspended in chloroform (40 ml) at 26° C. and is aerated with argon.

[0239] Diisopropylethylamine (1.5 mmol; 0.26 ml) is added dropwise through the septum.

[0240] Then, carbodiimidazole (6 mmol; 972 mg) is colloidally dissolved in chloroform (11 ml) and added dropwise through the septum.

[0241] This is stirred for 35 min at 26° C. Then, 2-oxa-6-azaspiro[3.3]heptane base (6 mmol; 594 mg) is added dropwise through the septum and stirred at 26° C.

[0242] After stirring for 2.5 h at 26° C., the formation of a product can be observed by TLC/LC-MS (see FIG. 5).

Example 7: Conversion into 2-oxa-6-azaspiro[3.3]heptyl-lysergic acid amide and iso-2-oxa-6-azaspiro[3.3]heptyl-lysergic acid amide

[0243] Conversion into 58% 2-oxa-6-azaspiro[3.3]heptyl-lysergic acid amide and iso-2-oxa-6-azaspiro[3.3]heptyl-lysergic acid amide.

[0244] At 312 nm, 31% of lysergic acid are still measurable. Therefore, stirring at 26° C. is continued for further 18 h.

[0245] After 2.5 h, the reaction mixture contains 58% isomer mixture and 31% starting material.

[0246] Then, the reaction mixture is diluted with chloroform (100 ml) and extracted with water (40 ml).

[0247] The organic phase is dried over magnesium sulfate and concentrated on the rotary evaporator.

[0248] 1.10 g of a yellow-brown oil is obtained, which still contains amounts of diisopropylethylamine.

[0249] The purification is done by means of a 100 g silica column using dichloromethane/methanol at 80:20 as an eluent. The crude product is dissolved in pure dichloromethane (8 ml) and applied. The column is run isocratically with dichloromethane/methanol at 80:20 as an eluent.

[0250] According to HPLC at 312 nm, fractions 3 and 4 contain 2-oxa-6-azaspiro[3.3]heptyl-lysergic acid amide at a purity of 97%. Fractions 5-9 contain both isomers at a ratio of 1:1.

[0251] Without previous isomer optimization, 0.32 g of a yellow oil is thus obtained.

[0252] This corresponds to a yield of 31%.

Example 8: Binding Inhibition by the LSD Derivatives of the Invention at Multiple Receptor Targets

Introduction

[0253] Four novel LSD derivatives according to the present invention (referred to as “Compound 1”, “Compound 2”, “Compound 3”, and “Compound 4”; structures depicted below) were screened at 10 μM at key receptors of interest in order to determine % binding inhibition. Targets were selected due to prior evidence that LSD binds these targets with moderate or high affinities in vitro, as well as their functional significance in the context of human health.

[0254] Generally, % inhibition results greater than 50% are interpreted as displaying a potentially significant interaction.

Compound 1 (2-oxa-6-azaspiro[3.3]heptyl-lysergic acid amide)

[0255] ##STR00030##

Compound 2 (6-(2-mono-fluoro-ethyl)-6-nor-lysergic acid diethylamide)

[0256] ##STR00031##

Compound 3 (lysergic acid mono-(trifluoroethyl)amide)

[0257] ##STR00032##

Compound 4 (lysergic acid monofluoro diethylamide)

[0258] ##STR00033##

Methods

5-HT.SUB.2A .Receptor

[0259] Human recombinant serotonin 5-HT.sub.2A receptors expressed in CHO-K1 cells were used in modified Tris-HCl buffer pH 7.4. A 30 μg aliquot of membrane protein was incubated with 0.5 nM [.sup.3H]Ketanserin for 60 minutes at 25° C. Non-specific binding was estimated in the presence of 1 μM Mianserin. Receptors were filtered and washed, the filters were then counted to determine [.sup.3H]Ketanserin specifically bound. Test compounds were screened at 10 μM.

5-HT.SUB.2B .Receptor

[0260] Human recombinant serotonin 5-HT.sub.2B receptor expressed in CHO-K1 cells were used to prepare membranes in modified Tris-HCl buffer pH 7.4. A 30 μg aliquot of membrane protein was incubated with 1.2 nM [.sup.3H]LSD for 60 minutes at 37° C. Non-specific binding was estimated in the presence of 10 μM serotonin. Membranes were filtered and washed, the filters were then counted to determine [.sup.3H]LSD specifically bound. Test compounds were screened at 10 μM.

5-HT.SUB.2C .Receptor

[0261] CHO-K1 cells stably transfected with a plasmid encoding the human recombinant serotonin 5-HT.sub.2c receptors were used to prepare membranes in modified Tris-HCl buffer pH 7.4. A 3.2 μg aliquot of membrane protein was incubated with 1.0 nM [.sup.3H]Mesulergine for 60 minutes at 25° C. Non-specific binding was estimated in the presence of 1 μM Mianserin. Membranes were filtered and washed, the filters were then counted to determine [.sup.3H]Mesulergine specifically bound. Test compounds were screened at 10 μM.

5-HT.SUB.1A .Receptor

[0262] Human recombinant serotonin 5-HT.sub.1A receptors expressed in CHO-K1 cells were used in modified Tris-HCl buffer pH 7.4. An 8 μg aliquot of membrane protein was incubated with 1.5 nM [.sup.3H]8-OH-DPAT for 60 minutes at 25° C. Non-specific binding was estimated in the presence of 10 μM metergoline. Receptors were filtered and washed, the filters were then counted to determine [.sup.3H]8-OH-DPAT specifically bound. Test compounds were screened at 10 μM.

D.SUB.1 .Receptor

[0263] Human recombinant dopamine D.sub.1 receptors expressed in CHO cells were used in modified Tris-HCl buffer pH 7.4. A 20 μg aliquot was incubated with 1.4 nM [.sup.3H]SCH-23390 for 120 minutes at 37° C. Non-specific binding was estimated in the presence of 10 μM (+)-butaclamol. Receptors were filtered and washed, the filters were then counted to determine [.sup.3H]SCH-23390 specifically bound. Test compounds were screened at 10 μM.

D.SUB.2S .Receptor

[0264] Human recombinant dopamine D.sub.2S receptors expressed in CHO cells were used in modified Tris-HCl buffer pH 7.4. A 15 μg aliquot was incubated with 0.16 nM [.sup.3H]Spiperone for 120 minutes at 25° C. Non-specific binding was estimated in the presence of 10 μM haloperidol. Receptors were filtered and washed, the filters were then counted to determine [.sup.3H]Spiperone specifically bound. Test compounds were screened at 10 μM.

D.SUB.2L .Receptor

[0265] Human recombinant dopamine D.sub.2L receptor expressed in CHO cells were used in modified Tris-HCl buffer pH 7.4. A 20 μg aliquot was incubated with 0.16 nM [.sup.3H]Spiperone for 120 minutes at 25° C. Non-specific binding was estimated in the presence of 10 μM haloperidol. Receptor proteins were filtered and washed, the filters were then counted to determine [.sup.3H]Spiperone specifically bound. Test compounds were screened at 10 μM.

D.SUB.3 .Receptor

[0266] Human recombinant dopamine D.sub.3 receptors expressed in CHO cells were used in modified Tris-HCl buffer pH 7.4. A 10 μg aliquot was incubated with 0.7 nM [.sup.3H]Spiperone for 120 minutes at 37° C. Non-specific binding was estimated in the presence of 25 μM S(−)-sulpiride. Receptors were filtered and washed, the filters were then counted to determine [.sup.3H]Spiperone specifically bound. Test compounds were screened at 10 μM.

D.SUB.47 .Receptor

[0267] CHO-K1 cells stably transfected with a plasmid encoding the human dopamine D.sub.4.7 receptor were used to prepare membranes in modified Tris-HCl buffer pH 7.4. A 60 μg aliquot of membrane was incubated with 1.5 nM [.sup.3H]Spiperone for 120 minutes at 25° C. Non-specific binding was estimated in the presence of 10 μM haloperidol. Membranes were filtered and washed, the filters were then counted to determine [.sup.3H]Spiperone specifically bound. Test compounds were screened at 10 μM.

A.SUB.2A .Receptor

[0268] Human recombinant adenosine A.sub.2A receptors expressed in human HEK-293 cells were used in modified Tris-HCl buffer pH 7.4. A 15 μg aliquot was incubated with 50 nM [.sup.3H]CGS-21680 for 90 minutes at 25° C. Non-specific binding was estimated in the presence of 50 μM NECA. Receptor were filtered and washed, the filters were then counted to determine [.sup.3H]CGS-21680 specifically bound. Test compounds were screened at 10 μM.

Analysis

[0269] Experimental results were expressed as a percent of control specific binding, via the equation:

(100−(measured specific binding×100)

[00001]$\left(100-(\frac{\mathrm{measured}\mathrm{specific}\mathrm{binding}}{\mathrm{control}\mathrm{specific}\mathrm{binding}}\times 100\right)$

[0270] For each target, data from two experimental replicates were averaged to generate a mean % inhibition.

Results

[0271] The experimental data obtained are summarized in the following Table 1.

TABLE-US-00002 TABLE 1 Compound 1 Compound 2 Compound 3 Compound 4 Target % inhibition % inhibition % inhibition % inhibition 5-HT.sub.2A 95.3 95.7 93.1 97.1 5-HT.sub.2B 96.8 90.5 94.9 97.9 5-HT.sub.2C 85.8 4.8 89.4 100.4 5-HT.sub.1A 96.3 100.2 100.6 100.5 D.sub.2S 83.1 0.6 64.2 92.1 D.sub.2L 78.8 7.9 63.1 92.7 D.sub.1 85.6 4.3 53.8 97.1 D.sub.3 94.5 14.2 94.7 97.7 D.sub.4.7 80.9 2.8 20.1 71.6 A.sub.2A −12.3 −2.5 −2.0 12.3 % inhibition of novel compounds at target receptors of interest. Higher % inhibition indicates a likely stronger ligand-receptor interaction.

[0272] These results demonstrate that the LSD derivatives according to the present invention, including Compound 1, Compound 2, Compound 3 and Compound 4, are advantageously selective ligands of the 5-HT.sub.2A receptor.

[0273] Moreover, compounds such as Compound 2 have been found to exhibit an advantageous receptor subtype selectivity. Thus, a very high % inhibition was observed for Compound 2 at the 5-HT.sub.2A and 5-HT.sub.1A receptors while no significant interaction was determined at the 5-HT.sub.2C, A.sub.2A or any dopamine receptors assayed. For Compound 3, significant interaction was observed at all 5-HT targets while no significant interaction was observed at the A.sub.2A or D.sub.47 receptors; the interaction with D.sub.3 showed a higher % inhibition than at other dopamine receptors, which may indicate a greater selectivity for D.sub.3 over other dopamine receptors. For all tested compounds, including Compound 1 and Compound 4, no significant interaction was determined at the A.sub.2A receptor.

[0274] These findings confirm that the LSD derivatives provided herein are particularly well suited for use in therapy, including for the treatment of serotonin 5-HT.sub.2A receptor associated diseases/disorders.

Example 9: Metabolization Study Using a Pooled Human Liver Microsome Assay

[0275] Pooled human liver microsome (pHLM) samples were prepared by adding novel LSD derivatives (“L1”, “L2”, “L3” or “L4”; structures shown in the schemes below) (approx. 1 mg/mL in ACN) solution to a reaction mixture (approx. 10 μg/mL final substrate concentration) consisting of pHLM, phosphate buffer, and deionized water. Incubation was performed for 30 minutes at 37° C. and stopped by the addition of ice-cold ACN. After centrifugation, the supernatant was diluted (1:10 for liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) and 1:2 for liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QToF-MS) analysis with mobile phase A/B (50/50, v/v). Two negative control samples were processed the same way, one with 2.5 μL phosphate buffer instead of pHLM and the second with 0.5 μL ACN instead of the substrate.

[0276] For the identification of tentative main metabolites, a Nexera X2 UHPLC (Shimadzu, Duisburg, Germany) coupled to a QTRAP® 5500 triple quadrupole linear ion trap mass spectrometer (SCIEX, Darmstadt, Germany) was utilized for analysis of pHLM. Phase I metabolism of the four novel LSD derivatives as well as a corresponding measurement of LSD as a reference were conducted.

[0277] As a result, for all four LSD derivatives similar metabolism derivatization reactions like hydroxylation, dihydroxylation, amine-demethylation and dealkylation of the amide moieties in analogy to the metabolism of LSD have been observed, as illustrated in the following schemes:

##STR00034##

##STR00035##

##STR00036##

##STR00037##

##STR00038##

[0278] It has thus been shown that the novel LSD derivatives according to the invention, including the compounds L1, L2, L3 and L4, undergo a similar metabolization in human liver microsomes as LSD, and do not give rise to toxic metabolites. These findings further confirm the suitability of the LSD derivatives provided herein to be used in therapy.