ERGOLINE DERIVATIVES AS ANALGESICS

Technical Field

The present invention relates to an analgic agent which comprises a clavine alkaloid derivative that exhibits substance P receptor antagonistic activity and is useful as a non-narcotic analgic agent.

Background Art

Substance P, which belongs to tachykinin peptides, like substance K (neurokinin A) and neurokinin B- is known to exhibit various physiological activities [Physiological Previews, Vol. 1- p. 1 (1991)]. Since substance P plays an important role as a neurotransmitter involved in the pain sensation of the unmyelinated sensation nerve projected to the posterior spinal dorsal root and as an inflammation mediator, substance P antagonists are used as analgic anti- inflammatory agents.

Setoclavine and agroclavine, represented by the above formulas, respectively, are clavine alkaloids produced by ergot fungus. Their production, isolation and structures are already reported [A. Hofmann et al., Helvetica Chimica Acta, Vol. 40, pp. 1358-1373 (1957)]. Concerning this class of low-molecular clavine alkaloids, there have been reports regarding the tumor cell growth inhibitory activity

of agroclavine [E. Eich et al., Biochemical Pharmacology, Vol. 33, pp. 523-526 (1984)] and its hypotensive and sedative actions [Folia Pharmacologica Japonica, Vol. 58, pp. 386-393, pp. 417-429 (1962)]. However, there is no report of its substance P antagonistic or analgic activity.

Disclosure of Invention

Analgic agents are necessary to patients with rheumatoid arthritis, neuralgia, osteoporosis, terminal cancer etc. for pain killing; non-narcotic analgic agent with low addictive property is required.

In view of these circumstances, the present inventors investigated non-narcotic analgic agent, and found that clavine alkaloid derivatives exhibit substance P receptor antagonistic activity, and exhibit analgic activity in various animal models. The inventors made further investigations based on these findings, and developed the present invention.

Accordingly, the present invention relates to: 1) An analgic agent which comprises a compound of the general formula:

wherein each of Ri and R ₂ is a hydrogen atom or an optionally substituted hydrocarbon group; R ₃ is a lower alkyl group; rings A and B may optionally be substituted; ring D is further substituted with an optionally substituted hydroxyl group and may optionally be

substituted with an oxo group, wherein a double bond is formed between the positions 8 and 9 or between the positions 9 and 10, or a pharmaceutically acceptable salt thereof;

2) An analgic agent according to the above paragraph 1, wherein the hydrocarbon group is a Cι_ιo alkyl group;

3) An analgic agent according to the above paragraph 1, wherein the substituted hydroxyl group is a hydroxyl group which is substituted with a C ₁ - ₇ hydrocarbon group;

10 4) An analgic agent according to the above paragraph 1, wherein the substituted hydroxyl group is a hydroxyl group which is substituted with a lower alkyl group;

5) An analgic agent according to the above paragraph 1, wherein the ring D is further substituted with a hydroxyl

15 group at the position 10, wherein a double bond is formed between the positions 8 and 9;

6) An analgic agent according to the above paragraph 1, wherein the ring D is further substituted with an optionally substituted hydroxyl group at the position 8,

20 wherein a double bond is formed between the positions 9 and 10.

7) An analgic agent according to the above paragraph 1, wherein the ring D is further substituted with a hydroxyl group which may optionally be substituted with a C ₁ - ₇ ng hydrocarbon group at the position 8, wherein a double bond is formed between the positions 9 and 10;

8) An analgic agent according to the above paragraph 1, wherein the analgic agent is for oral administration;

9) A compound of the general formula:

30

wherein each of R and R ₅ is a hydrogen atom or an optionally substituted hydrocarbon group; Re is a lower alkyl group; rings A and B may optionally be substituted; ring D is further substituted with an optionally substituted hydroxyl group and may optionally be substituted with an oxo group, wherein a double bond is formed between the positions 8 and 9 or between the positions 9 and 10; ring D is further substituted with a hydrocarbonoxy group containing 2 or more carbon atoms at the position 8 when a double bond is formed between the positions 9 and 10; ring D is further substituted with an optionally substituted hydroxyl group at the position 10 when a double bond is formed between the positions 8 and 9, and the free hydroxyl group at the position 10 and the hydrogen atom at the position 5 having the same orientation in the case that R ₄ is a hydrogen atom, each of R ₅ and Re is methyl group, or salt thereof;

10) a compound according to the above paragraph 9, wherein the hydrocarbon group is a Cι_ιo alkyl group;

11) A compound according to the above paragraph 9, wherein the hydrocarbonoxy group containing 2 or more carbon atoms is a C ₂ - ₆ alkoxy group;

12) A compound according to the above paragraph 9, wherein the substituted hydroxyl group is a hydroxyl group which is substituted with a C ₁ - ₇ hydrocarbon group;

13) A compound according to the above paragraph 9, wherein the substituted hydroxyl group is a lower alkoxy group;

14) A compound according to the above paragraph 9, wherein the ring D is further substituted with a C ₂ _ ₆ alkoxy group at the position 8, wherein a double bond is formed between the positions 9 and 10;

15) A compound according to the above paragraph 9, wherein the ring D is further substituted with a C ₂ - ₆ alkenyloxy group at the position 8, wherein a double bond is formed between the positions 9 and 10;

16) A compound according to the above paragraph 9, wherein

10 the ring D is further substituted with a C ₇ - ₁₃ aralkyloxy group at the position 8, wherein a double bond is formed between the positions 9 and 10;

17) A compound according to the above paragraph 9, wherein the compound is (5R,10R) and (5S,10S)-8,9-didehydro-6,8-

■ _j c dimethyl-10-ergolinol;

18) A compound according to the above paragraph 9, wherein the compound is (5R,8S)-9,10-didehydro-6,8-dimethyl-l- ethyl-8-ergolinol;

19) A compound according to the above paragraph 9, wherein 20 the compound is (5R,8S)-9,10-didehydro-6,8-dimethyl-l- propyl-8-ergolinol;

20) A compound according to the above paragraph 9, wherein the compound is (5R,8S)-9,10-didehydro-6,8-dimethyl-l- methoxymethyl-8-ergolinol;

25 21) A compound according to the above paragraph 9, wherein the compound is (5R,8S)-l-butyl-9,10-didehydro-6,8- dimethyl-8-ergolinol;

22) A compound according to the above paragraph 9, wherein the compound is (5R,8S)-9,10-didehydro-6,8-dimethyl-l-(2-

O hydroxypropyl)-8-ergolinol;

23) A compound according to the above paragraph 9, wherein the compound is (5R,8S)-9,10-didehydro-6-ethyl-8-methyl-8- ergolinol;

24) A compound according to the above paragraph 9, wherein 3 the compound is (5R,8S)-9,10-didehydro-8-methyl-6-propyl-8- ergolinol;

25) A compound according to the above paragraph 9, wherein the compound is (5R,8S)-6-cyclopropylmethyl-9,10-didehydro- 8-methyl-8-ergolinol;

26) An analgic agent as defined in the above paragraph 1, which is for pain killing;

27) Use of a compound or a salt thereof according to the above paragraph 1 to manufacture an analgic agent;

28) A process for producing a compound of the general formula (I) or a salt thereof, which comprises subjecting a compound of the general formula;

wherein each of R ₇ and Re is a hydrogen atom or an optionally substituted hydrocarbon group; Rg is a lower alkyl group; rings A and B may optionally be substituted, or a salt thereof to an intramolecular amidation and then reducing the resultant amido group, followed by subjecting a hydroxyl group to rearrangement and/or etherification, if necessary; 29) A process for producing a compound of the general formula (I) or a salt thereof, which comprises subjecting a compound of the general formula;

[IV]

wherein Rio is an optionally substituted hydrocarbon group; Rii is a lower alkyl group; X is a hydrogen atom or a hydrocarbon group; rings A and B may optionally be substituted, or a salt thereof to an N-alkylation; 30) A process for producing a compound of the general formula (I) or a salt thereof, which comprises subjecting a compound of the general formula;

wherein R ₁₂ is a hydrogen atom or an optionally substituted hydrocarbon group; R ₁₃ is a hydrogen atom or an optionally substituted hydrocarbon group; R ₁ is a lower alkyl group, R ₁₂ is an optionally substituted hydrocarbon group in the case that each of R ₁₃ and R ₁₄ is methyl group, or a salt thereof to an oxidation.

31) A method of alleviating pain in a patient, which comprises the step of administering to a patient in need of such treatment a pain alleviating effective amount of an analgic agent according to the above paragraph 1; and 32) A pharmaceutical composition for pain killing which comprises a compound according to the above paragraph 9 or a pharmaceutical acceptable salt thereof and a pharmaceutical carrier.

Best Mode for Carrying Out the Invention

With respect to the compound of general formula [I], ring D is further substituted with an optionally substituted hydroxyl group at the position 10 when a double bond is formed between the positions 8 and 9; ring D is further substituted with an optionally substituted hydroxyl

group at the position 8 when a double bond is formed between the positions 9 and 10. With respect to general formula [I], the hydrocarbon groups for Ri or R ₂ that may have a substituent include hydrocarbon groups having 1 to 20 carbon atoms. Such hydrocarbon groups include alkyl groups, alkenyl groups, alkynyl groups, aryl groups, aralkyl groups, cycloalkyl groups and cycloalkyl-alkyl groups.

Preferable alkyl groups include alkyl groups having 1

10 to 10 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, 1-methylbutyl, 2- methylbutyl, 1,2-dimethylpropyl, 1-ethylpropyl, hexyl, isohexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl,

1 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl,

3,3-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 1- ethylbutyl, 2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2- trimethylpropyl, 1-ethyl-l-methylpropyl, l-ethyl-2- methylpropyl, heptyl, isoheptyl, octyl, isooctyl, decyl,

2o isodecyl and so on. Of these alkyl groups, those having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl etc. are preferred, with greater preference given to those having 1 to 4 carbon atoms, such as methyl, ethyl,

25 propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl and so on.

Preferable alkenyl groups include alkenyl groups having 2 to 10 carbon atoms, such as vinyl, allyl, isopropenyl, 1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, gθ 1-methyl-l-propenyl, l-methyl-2-propenyl, 2-methyl-l- propenyl, 2-methyl-2-propenyl, 1-pentenyl, 2-pentenyl, 3- pentenyl, 4-pentenyl, 1-methyl-l-butenyl, 2-methyl-l- butenyl, 3-methyl-l-butenyl, l-methyl-2-butenyl, 2-methyl- 2-butenyl, 3-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-

35 hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-l-pentenyl, 2- methyl-1-pentenyl, 4-methyl-3-pentenyl, 2-ethyl-l-butenyl,

2-heptenyl, 2-octenyl, 2-decenyl and so on. Of these alkenyl groups, those having 2 to 5 carbon atoms, such as vinyl, allyl, 2-butenyl, 3-butenyl, isopropenyl, 2-methyl- 1-propenyl and 3-methyl-2-butenyl etc. are preferred, with greater preference given to those having 3 to 5 carbon atoms, such as allyl, 2-butenyl, 3-butenyl and 3-methyl-2- butenyl and so on.

Preferable alkynyl groups include alkynyl groups having 2 to 10 carbon atoms, such as ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, l-methyl-2- propynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, l-methyl-3-butynyl, 2-methyl-3-butynyl, 1-hexynyl, 2- hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, 2-heptynyl, 2- octynyl and 2-decynyl and so on. Of these alkynyl groups, those having 2 to 4 carbon atoms, such as ethynyl, 1- propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl and l-methyl-2-propynyl etc. are preferred, with greater preference given to those having 2 to 3 carbon atoms, such as ethynyl, 1-propynyl and 2-propynyl and so on. Preferable aryl groups include aryl groups having 6 to 14 carbon atoms, such as phenyl, tolyl, xylyl, biphenyl, anthracenyl, 1- or 2-naphthyl and 1-, 2-, 4-, 5- or 6- azurenyl and so on. Of these aryl groups, those having 6 to 8 carbon atoms, such as phenyl, tolyl and xylyl, etc. are preferred.

Preferable aralkyl groups include aralkyl groups having 7 to 20 carbon atoms, such as benzyl, phenethyl, 3- phenylpropyl, benzhydryl, trityl, triphenylethyl, (1- naphthyl)methyl and (2-naphthyl)methyl and so on. Of these aralkyl groups, benzyl, phenethyl and benzhydryl, etc. are preferred.

Preferable cycloalkyl groups include cycloalkyl groups having 3 to 7 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. Preferable cycloalkyl-alkyl groups include alkyl groups having 1 to 4 carbon atoms and substituted by the

above-mentioned cycloalkyl groups, such as cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl, cyclohexylbutyl and so on. Of these cycloalkyl-alkyl groups, cyclopropylmethyl is preferred.

Such hydrocarbon group residues may have 1 to 5 substituents at any possible positions thereon, which substituents are selected from:

(1) amino groups that may be substituted by mono- or di-Ci- ₄ alkyl groups (e.g., amino, methylamino, ethylamino, propylamino, isopropylamino, butylamino, dimethylamino, diethylamino, etc.),

(2) alkanoylamino groups having 1 to 6 carbon atoms (e.g., formylamino, acetylamino, propionylamino, butyrylamino, isobutyrylamino, valerylamino, isovalerylamino, pivaloylamino, hexanoylamino, etc.),

(3) aroylamino groups having 7 to 11 carbon atoms (e.g., benzoylamino, p-toloylamino, 1-naphthoylamino, 2- naphthoylamino, etc.),

(4) alkoxycarbonylamino groups having 2 to 7 carbon atoms (e.g., methoxycarbonylamino, ethoxycarbonylamino, propoxycarbonylamino, isopropoxycarbonylamino, tert- butoxycarbonylamino, etc.),

(5) aralkyloxycarbonylamino groups having 8 to 12 carbon atoms (e.g., benzyloxycarbonylamino, phenethyloxycarbonylamino, etc.),

(6) alkylsulfonylamino groups having 1 to 6 carbon atoms (e.g., methylsulfonylamino, ethylsulfonylamino, propylsulfonylamino, etc.),

(7) arylsulfonylamino groups having 6 to 12 carbon atoms (e.g., phenylsulfonylamino, tosylamino, etc.),

(8) hydroxyl groups,

(9) alkoxy groups having 1 to 4 carbon atoms (e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, etc.), (10) aryloxy groups having 6 to 10 carbon atoms (e.g., phenoxy, etc.).

(11) aralkyloxy groups having 7 to 12 carbon atoms (e.g., benzyloxy, etc.),

(12) alkanoyloxy groups having 1 to 6 carbon atoms (e.g., formyloxy, acetyloxy, propionyloxy, butyryloxy, isobutyryloxy, valeryloxy, isovaleryloxy, pivaloyloxy, hexanoyloxy, etc.),

(13) aroyloxy groups having 7 to 11 carbon atoms (e.g., benzoyloxy, p-toloyloxy, 1-naphthoyloxy, 2-naphthoyloxy, etc.) ,

(14) mercapto groups,

(15) alkylthio groups having 1 to 4 carbon atoms (e.g., methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, sec-butylthio, tert-butylthio, etc. ) ,

(16) arylthio groups having 6 to 10 carbon atoms (e.g., phenylthio, naphthylthio, etc.),

(17) alkylsulfinyl groups having 1 to 4 carbon atoms (e.g., methylsulfinyl, ethylsulfinyl, propylsulfinyl, etc.),

(18) arylsulfinyl groups having 6 to 10 carbon atoms (e.g., phenylsulfinyl, etc.),

(19) alkylsulfonyl groups having 1 to 4 carbon atoms (e.g., methylsulfonyl, ethylsulfonyl, propylsulfonyl, etc.),

(20) arylsulfonyl groups having 6 to 10 carbon atoms (e.g., phenylsulfonyl, tosyl, etc.),

(21) carboxyl groups,

(22) alkoxycarbonyl groups having 2 to 5 carbon atoms (e.g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl, etc.),

(23) aralkyloxycarbonyl groups having 8 to 13 carbon atoms (e.g., benzyloxycarbonyl, phenethyloxycarbonyl, etc.),

(24) aryloxycarbonyl groups having 7 to 11 carbon atoms (e.g., phenoxycarbonyl, etc. ) ,

(25) carbamoyl groups,

(26) mono- or di-Cι- ₄ alkylcarbamoyl groups (e.g., methylcarbamoyl, ethylcarbamoyl, propylcarbamoyl, butylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl, etc.).

(27) guanidyl groups that may be substituted for by a nitro group,

(28) nitro groups,

(29) nitrile groups (cyano groups),

(30) halogen atoms (e.g., iodine, bromine, chlorine, fluorine, etc.),

(31) methylene groups, and

(32) 3- to 6-membered heterocyclic groups or condensed heterocyclic groups thereof that may have 1 to 4 substituents selected from (a) halogen atoms (e.g., bromine, chlorine, fluorine, etc.), (b) alkyl groups having 1 to 4 carbon atoms (e.g., methyl, ethyl, propyl, isopropyl etc.) and (c) halogenophenoxy groups (e.g., o-, m- or p- chlorophenoxy, o-, m- or p-bromophenoxy, etc.) and that contain 1 to 4 heteroatoms selected from atoms of oxygen, sulfur, nitrogen etc., in addition to carbon atoms, (e.g., oxiranyl, 2- or 3-oxetanyl, 2- or 3-thienyl, 2- or 3-furyl,

2- or 3-tetrahydrofuryl, 2-, 3- or 4-tetrahydropyranyl, 3-, 4- or 5-pyrazolyl, 2-, 4- or 5-thiazolyl, 3-, 4- or 5- isothiazolyl, 2-, 4- or 5-oxazolyl, 3-, 4- or 5-isoxazolyl, 2-, 4- or 5-imidazolyl, 1,2,3- or 1,2,4-triazolyl, 1H- or 2H-tetrazolyl, 2-, 3- or 4-pyridyl, 2-, 4- or 5-pyrimidyl,

3- or 4-pyridazinyl, quinolyl, isoquinolyl and indolyl etc. ) . Preferable lower alkyl groups for R ₃ include C ₁ - ₆ alkyl groups, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, 1-methylbutyl, 2-methylbutyl, 1,2- dimethylpropyl, 1-ethylpropyl, hexyl, isohexyl, 1- methylpentyl, 2-methylpentyl, 3-methylpentyl, 1,1- dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 3,3- dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 1- ethylbutyl, 2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2- trimethylpropyl, 1-ethyl-l-methylpropyl, l-ethyl-2- methylpropyl and so on. Of these alkyl groups, those having 1 to 6 carbon atoms, such as methyl, ethyl, propyl,

isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl etc. are preferred, with greater preference given to alkyl groups having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert- butyl and so on.

Substituents that may be present on ring A include halogen atoms, thiol groups that may be substituted (e.g., alkylthio groups, arylthio groups, aralkylthio groups, etc.), hydroxyl groups that may be substituted (e.g., alkoxy groups, etc.), amino groups that may be substituted (e.g., alkylamino, arylamino, aralkylamino, etc.), carboxyl groups that may be esterified, carbamoyl groups that may be substituted (e.g., mono- or di-Cι- ₄ alkylcarbamoyl groups, etc.), nitro groups, nitrile groups and heterocyclic groups, as well as hydrocarbon groups defined for R_ and R ₂ above.

Substitutents that may be present at the position 2 on ring B include hydroxyl groups that may be substituted (e.g., alkoxy groups, etc.), amino groups that may be substituted (e.g., alkylamino, arylamino, aralkylamino, etc.), carboxyl groups that may be esterified, carbamoyl groups that may be substituted (e.g., mono- or di-Cι- ₄ alkylcarbamoyl groups, etc.), nitro groups, nitrile groups and heterocylclic groups, as well as hydrocarbon groups defined for Ri and R ₂ above.

Example halogen atoms include atoms of fluorine, chlorine, bromine and iodine, with preference given to atoms of chlorine, bromine etc.

Alkylthio groups include those having 1 to 6 carbon atoms, such as methylthio, ethylthio, propylthio, butylthio, tert-butylthio, hexylthio and so on.

Arylthio groups include those having 6 to 12 carbon atoms, such as phenylthio, tolylthio, naphthylthio, biphenylthio and so on.

Aralkylthio groups include those having 7 to 13 carbon atoms, such as benzylthio, phenethylthio, benzhydrylthio and so on.

Alkoxy groups include those having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, sec- butoxy, tert-butoxy, hexyloxy and so on.

Alkylamino groups include amino groups substituted by 1 or 2 substituents selected from alkyl groups having 1 to 6 carbon atoms, such as methyl groups, ethyl groups, propyl groups, butyl groups, hexyl groups and so on.

Arylamino groups include amino groups substituted by 1 or 2 substituents selected from aryl groups having 6 to 12 carbon atoms, such as phenyl groups, tolyl groups, naphthyl groups, biphenyl groups and so on. Aralkylamino groups include amino groups substituted for by 1 or 2 substituents selected from aralkyl groups having 7 to 13 carbon atoms, such as benzyl groups, phenethyl groups, benzhydryl groups and so on.

Example carboxyl groups that may be esterified include those that may be esterified by, for example, (1) alkyl groups in the hydrocarbon groups defined for Ri and R ₂ above, and (2) benzyl groups that may have 1 to 3 substituents selected from nitro groups, halogen atoms (e.g., fluorine, chlorine, bromine, iodine) and alkoxy groups having 1 to 4 carbon atoms (e.g., methoxy, ethoxy, propoxy, isopropoxy, sec-butoxy, tert-butoxy, etc.), with preference given to those that may be esterified by (1) alkyl groups having 1 to 4 carbon atoms, and (2) benzyl groups that may be substituted for by nitro or halogens (e.g., p-nitrobenzyl, p-bromobenzyl etc.).

Mono- or di-Cι- ₄ alkylcarbamoyl groups include carbamoyl groups substituted for by 1 or 2 substituents selected from the above-mentioned alkyl groups having 1 to 4 carbon atoms. Example heterocyclic groups include aromatic heterocyclic groups having 1 to 3 atoms of oxygen, sulfur

or nitrogen, such as 2- or 3-thienyl, 2- or 3-furyl, 1-, 2- or 3-pyrrolyl, 2- , 3- or 4-pyridyl, 2-, 4- or 5- pyrimidinyl, 2-, 4- or 5-oxazolyl, 2-, 4- or 5-thiazolyl, 3-, 4- or 5-pyrazolyl, 2-, 4- or 5-imidazolyl, 3- or 5- (1,2,4-oxadiazolyl) , 3- or 5-(l,2,4-thiadiazolyl) , 1,3,4- thiadiazolyl, 4- or 5-(l,2,3-thiadiazolyl) , 1,2,5- thiadiazolyl, 1,2,3-triazolyl and 1,2,4-triazolyl, etc.; and non-aromatic heterocyclic groups having 1 to 3 atoms of oxygen, sulfur or nitrogen, such as morpholinyl,

JO thiomorpholinyl, oxoimidazinyl, dioxotriazinyl, pyrrolidinyl, piperidyl, pyranyl, thiopyranyl, 1,4- oxazinyl, 1,4-thiazinyl, 1,3-thiazinyl, piperazinyl, pyrazinyl and so on.

Hydroxyl groups that may be substituted for and that _j are present as substituents on ring D are preferably hydroxyl groups that may be substituted by hydrocarbon groups. Hydrocarbon groups that may substitute on hydroxyl groups include the same hydrocarbon groups as those defined for Ri and R ₂ above. Preferably such hydrocarbon groups

2o include the above-mentioned alkyl groups, alkenyl groups, alkynyl groups, aryl groups, aralkyl groups, cycloalkyl groups and cycloalkyl-alkyl groups, etc., with greater preference given to hydrocarbon groups having 1 to 7 carbon atoms, such as alkyl groups having 1 to 6 carbon atoms,

25 alkenyl groups having 2 to 6 carbon atoms, alkynyl groups having 2 to 6 carbon atoms, phenyl, benzyl and so on.

Example hydroxyl groups substituted by such hydrocarbon groups include alkoxy groups having 1 to 10 carbon atoms, alkenyloxy groups having 2 to 10 carbon

30 atoms, alkynyloxy groups having 2 to 10 carbon atoms, aryloxy groups having 6 to 14 carbon atoms, aralkyloxy groups having 7 to 20 carbon atoms, cycloalkyloxy groups having 3 to 7 carbon atoms and C ₃ _ cycloalkyl-Cι- ₄ alkyloxy groups and so on.

35 Example preferable alkoxy groups having 1 to 10 carbon atoms include methoxy, ethoxy, propoxy, isopropoxy, butoxy,

isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, isopentyloxy, neopentyloxy, tert-pentyloxy, 1-methylbutoxy, 2-methylbutoxy, 1,2-dimethylpropoxy, 1-ethylpropoxy, hexyloxy, isohexyloxy, 1,1-dimethylbutoxy, 2,2- dimethylbutoxy, 3,3-dimethylbutoxy, 2-ethylbutoxy, heptyloxy, octyloxy, decyloxy and so on. Of these alkoxy groups, those having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, isopentyloxy, neopentyloxy, tert- pentyloxy, 1-methylbutoxy, 2-methylbutoxy, 1,2- dimethylpropoxy, 1-ethylpropoxy, hexyloxy, isohexyloxy, 1,1-dimethylbutoxy, 2,2-dimethylbutoxy, 3,3-dimethylbutoxy, 2-ethylbutoxy etc. are particularly preferable.

Example preferable alkenyloxy groups having 2 to 10 carbon atoms include vinyloxy, allyloxy, isopropenyloxy, 1- propenyloxy, 1-butenyloxy, 2-butenyloxy, 3-butenyloxy, 1- methyl-1-propenyloxy, l-methyl-2-propenyloxy, 2-methyl-l- propenyloxy, 2-methyl-2-propenyloxy, 1-pentenyloxy, 2- pentenyloxy, 3-pentenyloxy, 4-pentenyloxy, 1-methyl-l- butenyloxy, 2-methyl-l-butenyloxy, 3-methyl-l-butenyloxy, l-methyl-2-butenyloxy, 2-methyl-2-butenyloxy, 3-methyl-2- butenyloxy, 1-hexenyloxy, 2-hexenyloxy, 3-hexenyloxy, 4- hexenyloxy, 5-hexenyloxy, 1-methyl-l-pentenyloxy, 2-methyl- 1-pentenyloxy, 4-methyl-3-pentenyloxy, 2-ethyl-l- butenyloxy, 2-heptenyloxy, 2-octenyloxy, 2-decenyloxy and so on. Of these alkenyloxy groups, those having 3 to 6 carbon atoms, such as allyloxy, 2-butenyloxy, 3-butenyloxy, l-methyl-2-propenyloxy, 2-methyl-2-propenyloxy, 2- pentenyloxy, 3-pentenyloxy, 4-pentenyloxy, l-methyl-2- butenyloxy, 2-methyl-2-butenyloxy, 3-methyl-2-butenyloxy, 2-hexenyloxy, 3-hexenyloxy, 4-hexenyloxy, 5-hexenyloxy, 4- methyl-3-pentenyloxy, etc. are particularly preferable.

Example preferable alkynyloxy groups having 2 to 10 carbon atoms include ethynyloxy, 1-propynyloxy, 2- propynyloxy, 1-butynyloxy, 2-butynyloxy, 3-butynyloxy, 1- methyl-2-propynyloxy, 1-pentynyloxy, 2-pentynyloxy, 3-

pentynyloxy, 4-pentynyloxy, l-methyl-3-butynyloxy, 2- methyl-3-butynyloxy, 1-hexynyloxy, 2-hexynyloxy, 3- hexynyloxy, 4-hexynyloxy, 5-hexynyloxy, 2-heptynyloxy, 2- octynyloxy, 2-decynyloxy and so on. Of these alkynyloxy groups, those having 2 to 6 carbon atoms, such as ethynyloxy, 1-propynyloxy, 2-propynyloxy, 1-butynyloxy, 2- butynyloxy, 3-butynyloxy, l-methyl-2-propynyloxy, 1- pentynyloxy, 2-pentynyloxy, 3-pentynyloxy, 4-pentynyloxy, l-methyl-3-butynyloxy, 2-methyl-3-butynyloxy, 1-hexynyloxy, 2-hexynyloxy, 3-hexynyloxy, 4-hexynyloxy, 5-hexynyloxy, etc. are particularly preferable.

Example preferable aryloxy groups having 6 to 14 carbon atoms include phenyloxy, tolyloxy, xylyloxy, biphenyloxy, anthracenyloxy, 1- or 2-naphthyloxy, 1-, 2-, 4-, 5- or 6-azurenyloxy, etc., with preference given to those having 6 to 8 carbon atoms, such as phenyloxy, tolyloxy, xylyloxy and so on.

Example preferable aralkyloxy groups having 7 to 20 carbon atoms include benzyloxy, phenethyloxy, 3- phenylpropyloxy, diphenylmethyloxy, triphenylethyloxy, (1- naphthyl)methyloxy, (2-naphthyl)methyloxy, etc., with greater preference given to those having 7 to 13 carbon atoms, such as benzyloxy, phenethyloxy, diphenylmethyloxy, etc. are particularly preferable. Example preferable cycloalkyloxy groups having 3 to 7 carbon atoms include cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy and so on.

Example preferable C ₃ .- ₇ cycloalkyl-Cι- ₄ alkyloxy groups include cyclopropylmethoxy, cyclopentylmethoxy, cyclohexylmethoxy, cyclohexylbutoxy and so on.

With respect to general formula [I] above, Ri is preferably a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, more preferably a hydrogen atom; R ₂ is preferably an alkyl group having 1 to 10 carbon atoms or a C ₃ - ₇ cycloalkyl-Cι- ₄ alkyl group; R ₃ is preferably an alkyl group having 1 to 6 carbon atoms, more preferably an alkyl

group having 1 to 4 carbon atoms; preferably ring D is substituted with a hydroxyl group at the position 10 when a double bond is formed between the positions 8 and 9; or ring D is substituted with a hydroxyl group that may be substituted with a hydrocarbon group having 1 to 7 carbon atoms at the position 8 when a double bond is formed between the positions 9 and 10.

Salts of compounds represented by general formula [I] include physiologically acceptable salts. Such salts include acid adduct salts, salts with inorganic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid etc., and salts with organic acids, such as acetic acid, oxalic acid, succinic acid, trifluoroacetic acid and so on. Compound [I] (or salt thereof) is used as a non- narcotic analgic agent, in mixture with a pharmacologically acceptable carrier, by a method in common use in the relevant field. The analgic agent of the present invention is provided as a parenteral or oral preparation. Example parenteral preparations include injections, drip infusions, solutions, suspensions, suppositories and so on. Example oral preparations include capsules, tablets, syrups, powders, granules and so on.

In producing parenteral preparations, e.g., injections, compound [I] (or salt thereof) may be mixed with isotonizing agents (e.g., glucose, sorbitol, mannitol, sodium chloride etc.), preservatives (e.g., benzyl alcohol, chlorobutanol, methyl p-hydroxybenzoate, etc.), anticoagulants (e.g., dextran sulfate, heparin, etc.), dissolving aids (e.g., cyclodextrins, Tween, etc.), stabilizers (e.g., polyethylene glycol, polylactic acid, etc.) and other additives. These additives are dissolved in commonly used aqueous diluents, such as aqueous solutions of glucose, physiological saline, Ringer's solutions, nutrient supplements and so on. Oral preparations may be supplemented with additives, such as

excipients, binders, disintegrating agents, lubricants, colorants, correctives, stabilizers and so on.

With low toxicity, these preparations can be safely used in mammals, such as human, bovine, pig and so on. These preparations are orally or parenterally administered. Although doses for human use varies with diseases, ages of individual patients and states of disease, it is preferable that the preparation be used to treat disease at doses of about 0.5 to 500 mg, more preferably about 10 to 200 mg, in 1 to 3 portions, daily, for an adult weighing 50 kg, based on the content of compound [I] (or salt thereof).

The compounds represented by general formula [II], falling within the scope of general formula [I] above, are new compounds not described in a literature.

With respect to general formula [II], the hydrocarbon group for R or R ₅ and the lower alkyl group for Re are exemplified by the same such groups as those defined for general formula [I] above.

The hydrocarbonoxy group having 2 or more carbon atoms that is present as a substituent at the position 8 on ring D is preferably a hydrocarbonoxy group having 2 to 20 carbon atoms. Such hydrocarbonoxy groups include alkoxy groups having 2 to 10 carbon atoms, alkenyloxy groups having 2 to 10 carbon atoms, alkynyloxy groups having 2 to 10 carbon atoms, aryloxy groups having 6 to 14 carbon atoms, aralkyloxy groups having 7 to 20 carbon atoms, cycloalkyloxy groups having 3 to 7 carbon atoms and C ₃ - ₇ cycloalkyl-Cι- ₄ alkyloxy groups.

Preferable alkoxy groups having 2 to 10 carbon atoms include ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, isopentyloxy, neopentyloxy, tert-pentyloxy, 1-methylbutoxy, 2- methylbutoxy, 1,2-dimethylpropoxy, 1-ethylpropoxy, hexyloxy, isohexyloxy, 1,1-dimethylbutoxy, 2,2- dimethylbutoxy, 3,3-dimethylbutoxy, 2-ethylbutoxy, heptyloxy, octyloxy, decyloxy and so on. Of these alkoxy

groups, those having 2 to 7 carbon atoms, such as ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert- butoxy, pentyloxy, isopentyloxy, neopentyloxy, tert- pentyloxy, 1-methylbutoxy, 2-methylbutoxy, 1,2- dimethylpropoxy, 1-ethylpropoxy, hexyloxy, isohexyloxy, 1,1-dimethylbutoxy, 2,2-dimethylbutoxy, 3,3-dimethylbutoxy, 2-ethylbutoxy, heptyloxy, etc. are particularly preferable, with greater preference given to those having 2 to 4 carbon atoms, such as ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy and so on.

Preferable alkenyloxy groups having 2 to 10 carbon atoms include vinyloxy, allyloxy, isopropenyloxy, 1- propenyloxy, 1-butenyloxy, 2-butenyloxy, 3-butenyloxy, 1- methyl-1-propenyloxy, l-methyl-2-propenyloxy, 2-methyl-l- propenyloxy, 2-methyl-2-propenyloxy, 1-pentenyloxy, 2- pentenyloxy, 3-pentenyloxy, 4-pentenyloxy, 1-methyl-l- butenyloxy, 2-methyl-l-butenyloxy, 3-methyl-l-butenyloxy, l-methyl-2-butenyloxy, 2-methyl-2-butenyloxy, 3-methyl-2- butenyloxy, 1-hexenyloxy, 2-hexenyloxy, 3-hexenyloxy, 4- hexenyloxy, 5-hexenyloxy, 1-methyl-l-pentenyloxy, 2-methyl- 1-pentenyloxy, 4-methyl-3-pentenyloxy, 2-ethyl-l- butenyloxy, 2-heptenyloxy, 2-octenyloxy, 2-decenyloxy and so on. Of these alkenyloxy groups, those having 3 to 7 carbon atoms, such as allyloxy, 2-butenyloxy, 3-butenyloxy, l-methyl-2-propenyloxy, 2-methyl-2-propenyloxy, 2- pentenyloxy, 3-pentenyloxy, 4-pentenyloxy, l-methyl-2- butenyloxy, 2-methyl-2-butenyloxy, 3-methyl-2-butenyloxy, 2-hexenyloxy, 3-hexenyloxy, 4-hexenyloxy, 5-hexenyloxy, 4- methyl-3-pentenyloxy, 2-heptenyloxy, etc. are particularly preferable, with greater preference given to those having 3 to 5 carbon atoms, such as allyloxy, 2-butenyloxy, 3- butenyloxy, 3-methyl-2-butenyloxy and so on.

Preferable alkynyloxy groups having 2 to 10 carbon atoms include ethynyloxy, 1-propynyloxy, 2-propynyloxy, 1- butynyloxy, 2-butynyloxy, 3-butynyloxy, l-methyl-2- propynyloxy, 1-pentynyloxy, 2-pentynyloxy, 3-pentynyloxy,

4-pentynyloxy, l-methyl-3-butynyloxy, 2-methyl-3- butynyloxy, 1-hexynyloxy, 2-hexynyloxy, 3-hexynyloxy, 4- hexynyloxy, 5-hexynyloxy, 2-heptynyloxy, 2-octynyloxy, 2- decynyloxy and so on. Of these alkynyloxy groups, those having 2 to 7 carbon atoms, such as ethynyloxy, 1- propynyloxy, 2-propynyloxy, 1-butynyloxy, 2-butynyloxy, 3- butynyloxy, l-methyl-2-propynyloxy, 1-pentynyloxy, 2- pentynyloxy, 3-pentynyloxy, 4-pentynyloxy, l-methyl-3- butynyloxy, 2-methyl-3-butynyloxy, 1-hexynyloxy, 2- hexynyloxy, 3-hexynyloxy, 4-hexynyloxy, 5-hexynyloxy, 2- heptynyloxy, etc. are particularly preferable, with greater preference given to those having 3 to 4 carbon atoms, such as 2-propynyloxy, 2-butynyloxy and so on.

Preferable aryloxy groups having 6 to 14 carbon atoms include phenyloxy, tolyloxy, xylyloxy, biphenyloxy, anthracenyloxy, 1- or 2-naphthyloxy, 1-, 2-, 4-, 5- or 6- azurenyloxy and so on. Of these aryloxy groups, those having 6 to 8 carbon atoms, such as phenyloxy, tolyloxy, xylyloxy, etc., are particularly preferable. Preferable aralkyloxy groups having 7 to 20 carbon atoms include benzyloxy, phenethyloxy, 3-phenylpropyloxy, diphenylmethyloxy, triphenylethyloxy, (1- naphthyl)methyloxy, (2-naphthyl)methyloxy and so on. Of these aralkyloxy groups, those having 7 to 13 carbon atoms, such as benzyloxy, phenethyloxy, diphenylmethyloxy, etc. are particularly preferable.

Preferable cycloalkyloxy groups having 3 to 7 carbon atoms include cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy and so on. Preferable C ₃ - ₇ cycloalkyl-Cι_ ₄ alkyloxy groups include cyclopropylmethoxy, cyclopentylmethoxy, cyclohexylmethoxy, cyclohexylbutoxy and so on.

With respect to general formula [II] above, R ₄ is preferably a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, more preferably a hydrogen atom; R ₅ is preferably an alkyl group having 1 to 10 carbon atoms or a

C ₃ - ₇ cycloalkyl-Cι_ ₄ alkyl group; Re is preferably an alkyl group having 1 to 6 carbon atoms, more preferably an alkyl group having 1 to 4 carbon atoms; preferably ring D is substituted with a hydrocarbonoxy group having 2 to 10 carbon atoms at the position 8, wherein a double bond is formed between the positions 9 and 10.

With respect to general formula [II], the substituent that may be present on rings A and B is exemplified by the same such substituent as defined for the general formula [I] above.

Salts of compounds represented by general formula [II] include the same salts as those of compounds represented by general formula [I].

With respect to general formula [III] above, R ₇ , Re and Rg respectively have the same definitions as those of Ri, R ₂ and R ₃ for general formula [I] above.

With respect to general formula [III], the substituent that may be present on rings A and B is exemplified by the same such substituent as defined for the general formula [I] above.

Compounds of general formula [III] other than those having a hydrogen atom for R ₇ and a methyl group for Re and R ₉ , (hereinafter referred to as compound [Ilia]), (or salt thereof) are produced from rugulovasine by known methods. Rugulovasine has been reported as produced by Penicillium concavoruqulosum by Abe et al (Japanese published examined patent application 19588/1971). For example, compound [Ilia] (or salt thereof) is produced by exposing rugulovasine (or salt thereof) to microbial oxidase, and subjecting the resulting rugulovamine to oxidation and substitution.

Any microorganism can be used to produce rugulovamine, as long as it is capable of oxidizing rugulovasine, or salt thereof. Preferable microorganisms include actinomycetes belonging to the genus Saccharothrix, Streptomyces,

Actinomyces or Actinoplanes. Preferable strains include

Microbial oxidation

Oxidation

_, . . Substitution _Λ , _____ ,

Rugulovamine ► Compound [IHa]

Saccharothrix mutabilis subsp. capreolus IFO 12847, Streptomyces varsoviensis IFO 13093, Streptomyces viridifaciens IFO 13352, Actinomyces streptomycini IFO 12918, Actinoplanes brasiliensis IFO 13938 and so on.

Saccharothrix mutabilis subsp. capreolus IFO 12847, Streptomyces varsoviensis IFO 13093, Streptomyces viridifaciens IFO 13352, Actinomyces streptomycini IFO

12918 and Actinoplanes brasiliensis IFO 13938 are listed in the List of Cultures, 9th edition, 1992, issued by the Institute for Fermentation, Osaka (IFO), and are available from IFO. Although the medium used to culture these microorganisms may be liquid or solid, as long as it contains available nutrients, it is preferable to use a liquid medium in the case of large-scale culture. The medium is supplemented with carbon sources, nitrogen sources, minerals and trace nutrient sources that are assimilatable by the microorganisms. Example carbon sources include glucose, lactose, maltose, dextrin, starch, glycerol, sorbitol, oils and fats (e.g., soybean oil, lard, chicken oil, etc.), n-paraffin and so on. Example nitrogen sources include meat extract, yeast extract, soybean flour, corn steep liquor, peptone, cottonseed oil, blackstrap molasses, urea and ammonium salts (e.g., ammonium sulfate, ammonium chloride, etc.) and so on.

In addition, salts including sodium, potassium, calcium, magnesium, etc., salts of metals such as iron,

manganese, zinc, cobalt, nickel, etc., salts such as phosphates, borates, etc., and salts of organic acids such as acetic acid, propionic acid, oxalic acid, etc., are used as appropriate. Amino acids (e.g., glutamic acid, aspartic acid, alanine, lysine, methionine, proline, etc.), peptides (e.g., dipeptides, tripeptides, etc.), vitamins (e.g., vitamin Bi, vitamin B ₂ , vitamin Bβ, nicotinic acid, vitamin B ₁₂ , vitamin C, etc.), nucleic acids (e.g., purine, pyrimidine, derivatives thereof and so on) etc. may also be contained.

Inorganic acids (e.g., hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, boric acid, etc.), organic acids (e.g., acetic acid, oxalic acid, citric acid, tartaric acid, etc.), alkalis (e.g., sodium hydroxide, potassium hydroxide, sodium carbonate, etc.), buffers (e.g., sodium dihydrogen phosphate, disodium hydrogen phosphate and so on), etc. may be added to regulate medium pH. Appropriate amounts of oils and fats (e.g., soybean oil, lard, chicken oil, etc.), surfactants, etc. may be added for the purpose of defoaming.

In liquid culturing, medium pH is preferably nearly neutral, with preference given to pH about 5 to 8. Culturing temperature is preferable about 20 to 37 ^β C. Culturing time is preferably about 6 to 96 hours, more preferably about 12 to 72 hours.

The methylamino group of rugulovasine is converted by the oxidase of the microorganism into an amino group.

Oxidase is used as such or in enzyme solution. The enzyme solution may be used as is for the above-described culture broth, or as a solution containing powdered crude enzyme prepared by adding acetone, etc. to the culture supernatant obtained by centrifugation of culture broth. In the present invention, it is preferable to use a culture broth. Also, the enzyme solution may be supplemented with coenzymes, such as nicotinamide adenine dinucleotide

(NAD ⁺ ), phosphate ester thereof (NADP ⁺ ) and reduction products thereof (NADH and NADPH) , dehydrogenases, such as D-glucose-6-phosphate dehydrogenase and glycerol-3- phosphate dehydrogenase, and inorganic salts, such as those of halogenated alkaline earth metals such as magnesium chloride and so on.

In reacting rugulovasine in the enzyme solution, the starting concentration is preferably about 50 μg/ml to about 2 mg/ml, more preferably about 100 μg/ml to about 1 mg/ml. Reaction temperature is preferably about 18°C to about 42 ^β C, more preferably about 24°C to 37°C. Reaction time is preferably about 1 minute to about 50 hours, more preferably about 5 minutes to about 30 hours.

A method of collecting rugulovamine or salt thereof from the reaction mixture is described below.

Since rugulovamine is fat-soluble under alkaline conditions, ordinary methods based on this property can be used. For example, (1) the enzyme reaction mixture is filtered after adding a filter aid, etc., or centrifuged; the separated solid is removed. The filtrate or supernatant thus obtained is adjusted to pH about 5 to about 11, preferably about 6 to about 10. Then a water- immiscible organic solvent (e.g., chloroform, ethyl acetate, methyl isobutyl ketone, isobutanol, etc.) is added to extract rugulovamine. The extract thus obtained is washed with an aqueous inorganic substance (e.g., aqueous sodium bicarbonate, aqueous sodium carbonate, etc.) or water and concentrated to give crude material containing the desired compound. Alternatively, (2) crude rugulovamine is collected from the enzyme reaction mixture or the filtrate obtained as described above, using a carrier. For eluting ruglovamine from the carrier to which ruglovamine in the enzyme reaction mixture is adsorbed, a mixed solvent is used that comprises an appropriate organic solvent (e.g., acetone, acetonitrile, methanol, etc.) and water with or without an appropriate amount of acid (e.g.,

hydrochloric acid, sulfuric acid, etc.). After removal of organic solvent, the eluted fraction is treated by the above-described solvent extraction method, to extract the desired compound. The extract thus obtained is concentrated to give a crude material containing rugulovamine.

Useful carriers include commonly used inorganic or organic carriers, such as activated charcoal for chromatography (produced by Takeda Chemical Industries, Ltd., Japan), silica gel [e.g., Kieselgel 60 (produced by E. Merck, Germany) etc.], microcrystalline cellulose [e.g., Avicel (produced by Asahi Chemical Industry, Co., Ltd., Japan), Funacel (produced by Funakoshi Co., Ltd., Japan) etc.], adsorptive resins [e.g., Diaion HP-20 or SP-207 (produced by Mitsubishi Kasei, Japan), Amberlite XAD-I or II (produced by Rohm & Haas, USA), etc.], molecular sieve resins [e.g., Sephadex LH-20 (produced by Pharmacia, Sweden)] and so on. Of these carriers, adsorptive resins are preferred. Various chromatography techniques can be advantageously used to further the crude material and obtain pure rugulovamine or a salt thereof. For example, column chromatography can be used with commonly used inorganic or organic carriers, such as activated charcoal for chromatography, silica gel, microcrystalline cellulose, adsorptive resins, cation exchange resins [e.g., Amberlite IR-120, IRC-50 or CG-50 (produced by Rohm & Haas, USA), Dowex 50W (produced by Dow Chemical, USA), Diaion SK1A (produced by Mitsubishi Kasei, Japan), etc.], ion exchange Sephadex [e.g., CM-Sephadex (produced by Pharmacia,

Sweden), etc.] and molecular sieve resins and so on. To elute ruglovamine from the carrier, an appropriate organic solvents (e.g., n-hexane, chloroform, dichloroethane, toluene, ethyl acetate, acetone, methanol, diethylamine, dipropylamine, etc.), selected according to the kind and nature of the carrier used; alone or a mixture thereof can

be used. It is also possible to use mixed solvents comprising an appropriate ratio of a water-miscible organic solvent (e.g., methanol, ethanol, acetone, acetonitrile, etc.) and water, aqueous alkali (e.g., sodium hydroxide, potassium hydroxide, sodium hydrogen carbonate, etc.), aqueous acid (e.g., hydrochloric acid, acetic acid, formic acid, phosphoric acid, etc.), aqueous salt (e.g., saline, acetate buffer, phosphate buffer, etc.), or the like.

Preparative high performance liquid chromatography (HPLC) can also be used to further purify the crude material and obtain the desired compound. As carriers, octadecyl silane (hereinafter abbreviate as ODS) or silica gel carriers can be used advantageously. In the case of ODS, for instance, methanol or a mixture of acetonitrile and aqueous salt can be used advantageously. The eluate containing the desired compound is extracted with an appropriate water-immiscible organic solvent; the resulting extract thus obtained is concentrated to dryness to give the pure compound. Since rugulovamine is a basic substance, it can be converted into a physiologically acceptable salt by reaction with acid, by per se known method. Example acids include organic acids (e.g., ethylsuccinic acid, lactobionic acid, oxalic acid, succinic acid, citric acid, lactic acid, acetic acid, methanesulfonic acid, etc.) and inorganic acids (e.g., sulfuric acid, hydrochloric acid, phosphoric acid, etc.).

The rugulovamine thus obtained is sequentially subjected to oxidation and substitution, or substitution, oxidation and substitution, to give compound [Ilia].

Oxidation is carried out by reacting rugulovamine or salt thereof with an oxidizing agent or a halogenating agent. The methyl group of 7-lactone ring of rugulovamine is converted with oxidation into a hydroxy methyl group or a halomethyl group.

Example oxidizing agents for this oxidation include selenium derivatives (e.g., selenium oxide, etc.), peracids (e.g., peracetic acid, performic acid, perbenzoic acid, m- chloroperbenzoic acid, etc.), chromic acids (e.g., chromic anhydride, chromic acid-sulfuric acid, chromic acid-acetic acid, etc.), chromates (e.g., sodium chromate, potassium chromate, pyridinium chromate, pyridinium chlorochromate etc.), dichromates (e.g., potassium dichromate, sodium dichromate, pyridinium dichromate, etc.), permanganates (e.g., potassium permanganate, sodium permanganate, etc.), perhalic acids (e.g., periodic acid, perbromic acid, perchloric acid, etc.), perhalates (e.g., sodium periodate, potassium periodate, sodium perchlorate, etc.), N- haloimides (e.g., N-bromosuccinimide, N-chlorosuccinimide, N-bromomaleimide, N-chloromaleimide, etc.), N-haloamides

(e.g., N-bromoacetamide, etc.), peracid esters (e.g., tert- butyl perbenzoate, tert-butyl m-chloroperbenzoate, etc.) and peroxides (e.g., hydrogen peroxide, tert-butyl hydroperoxide, etc.). Example halogenating agents for this halogenation include hydrogen halide (e.g., hydrogen chloride, hydrogen bromide, hydrogen iodide, etc.), phosphorus halide (e.g., phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, phosphorus tribromide, phosphorus pentabromide, etc.), organophosphorus compounds (e.g., triphenylphosphine-carbon tetrachloride, triphenylphosphine-bromine, triphenylphosphine-N- bromosuccinimide, triphenylphosphine-methyl iodide, etc.), thionyl chloride, oxalyl chloride, phosgen and so on. In this oxidation, an oxidizing agent or a halogenating agent is normally used at about 0.2 to about 20 mol, preferably at about 1 to about 10 mol, especially preferably at about 1 to about 3 mol per mol of rugulovamine.

This oxidation reaction is normally carried out in a solvant that does not adversely affect the oxidation reaction.

Such solvents include water, alcohols (e.g., methanol, ethanol, tert-buthanol, etc.), sulfoxides (e.g., dimethyl sulfoxide, etc.), halogenated hydrocarbons (e.g., chloroform, dichloromethane, carbon tetrachloride, 1,2- dichloroethane, chlorobenzene, etc.), ethers (e.g., diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, etc.), nitriles (e.g., acetonitrile, etc.), esters (e.g., ethyl acetate, ethyl formate, tert-butyl acetate, etc.), amides (e.g., formamide, dimethylformamide, dimethylacetoamide, etc.), ketones (e.g., acetone, methyl ethyl ketone, methyl isobutyl ketone, etc.), hydrocarbons (e.g., hexane, benzene, toluene, etc.), aromatic organic bases (e.g., pyridine, 2,4,6-trimethylpyridine, picoline, 2,4,6- lutidine, diazabicycloundecene, etc.) and so on; or an appropriate mixture thereof.

Of these solvents, water, amides (e.g., formamide, dimethylformamide, dimethylacetoamide, etc.), ketones (e.g., acetone, methyl ethyl ketone, methyl isobutyl ketone, etc.) are preferable.

Although reaction temperature is not subject to limitation as long as the reaction proceeds, the reaction is normally from about -50 to about 150°C, preferably about -30 to about 100°C especially preferably about 0 to about 100°C. Reaction time is normally from about 2 minutes to 48 hours, preferably about 5 minutes to about 10 hours, depending on reaction temperature and kind of solvent. With respect to protection of functional groups not involved in the reaction, protective groups therefore, elimination of the protective groups etc., known groups or known means can be selected as appropriate.

Substitution reactions include substitution at rugulovamine amino group or imino group (hereinafter referred as N-substitution) and substitution at the

hydroxyImethyl group, halomethyl group etc. resulting from oxidation of the methyl group on the y-lactone ring of rugulovamine (hereinafter referred as C-substitution) .

N-substitution is carried out by reacting the starting material with a compound having a leaving group in the presence of a base.

Example a compound having a leaving group preferably include halides, such as iodides (e.g., methyl iodide, ethyl iodide, propyl iodide, isopropyl iodide, butyl iodide, pentyl iodide, hexyl iodide, iodoacetic acid, iodoacetamide, iodoacetonitrile, methyl iodoacetate, tert- butyl iodoacetate, sodium iodoacetate, 2-iodo-l,l,l- trifluoromethane, etc.), bromides (e.g., ethyl bromide, propyl bromide, isopropyl bromide, butyl bromide, isobutyl bromide, tert-butyl bromide, pentyl bromide, hexyl bromide, allyl bromide, benzyl bromide, 2,2-diethoxyethyl bromide, tetrahydrofurfuryl bromide, cyclopropylmethyl bromide, epibromohydrin, triphenylmethyl bromide, etc.), chlorides (e.g., methoxymethyl chloride, methoxyethoxymethyl chloride, methylthiomethyl chloride, tetrahydropyranyl chloride, etc.), fluorides (e.g., 2,4-dinitrofluorobenzene, etc.), sulfonic acid esters such as p-toluenesulfonic acid esters (e.g., methyl p-toluenesulfonate, ethyl p- toluenesulfonate, etc.); methanesulfonic acid esters (e.g., methyl methanesulfonate, butyl methanesulfonate, etc.); trifluoromethanesulfonic acid esters (e.g., ethyl trifluoromethanesulfonate, etc.) and so on, sulfuric acid esters (e.g., dimethyl sulfate, diethyl sulfate, etc.), epoxides (e.g., propylene oxide, glycidol, etc.) and so on. Of these compounds, iodides, bromides, chlorides, and sulfonic acid esters are prepared from corresponding alcohols by the halogenation as described above, or sulfonic acid esterification and so on.

Preferable example bases include alkali metal hydrides (e.g., sodium hydride, potassium hydride, etc.), alkaline earth metal hydrides (e.g., calcium hydride, etc.), alkali

metal alkoxides (e.g., sodium methoxide, sodium ethoxide, potassium tert-butoxide, etc.), inorganic bases (e.g., sodium hydrogencarbonate, sodium carbonate, potassium hydrogencarbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, calcium hydroxide, etc.), organic bases (e.g., aromatic bases such as pyridine, 2,4,6- trimethylpyridine, picoline, 4-dimethylaminopyridine, 2,6- lutidine, diazabicycloundecene, etc.; tertiary amines such as triethylamine, dimethylaniline, etc.), alkali metals (e-g« sodium, potassium), metal hydrides (e.g., lithium aluminum hydride, lithium borohydride, diisobutylaluminum hydride, etc.), alkyl lithiums (e.g., methyl lithium, butyl lithium, etc.) and alkali metal amides (e.g., lithium diisopropylamide, lithium bistrimethylsilylamide, sodium bistrimethylamide, etc.). Of these bases, alkali metal hydrides, alkali metal alkoxides, alkali metals, alkyl lithiums, alkali metal amides, etc. are more preferably. Of these bases, liquid ones may be used as solvents.

In this reaction, a compound having a leaving group is normally used at about 1 to about 40 mol, preferably about 1 to about 10 mol per mol of starting compound. In this reaction, a base is normally used at about 1 to about 50 mol, preferably about 1 to about 10 mol per mol of starting material compound. This reaction is normally carried out in a solvent that does not adversely affect the reaction. Such solvents include the same solvents as those used for the above-described halogenation. Although reaction temperature is not subject to limitation as long as the reaction proceeds, the reaction is normally carried out at about -70 to about 150°C, preferably about 0 to about 80°C, especially preferably about 0 to about 60°C. Reaction time is normally about 2 minutes to about 96 hours, preferably about 2 minutes to about 48 hours, especially preferably about 10 minutes to 24 hours. With respect to protection of functional groups not involved in the reaction, protective groups therefor,

elimination of the protective groups etc., known groups or known means can be selected as appropriate.

C-substitution is carried out by reacting the hydroxymethyl group, halomethyl group, or the like, resulting from oxidation, with an alkyl anion or an equivalent thereof. In this reaction the hydroxyl group resulting from the above-described oxidation reaction be converted into a corresponding leaving group, such as a halogen atom (e.g., fluorine, chlorine, bromine, iodine), sulfonate (e.g., toluenesulfonate, benzenesulfonate, etc.), or the like, and then be subjected to the C-substitution.

Halogenation is carried out by reacting hydroxyrugulovamine, resulting from the above-described oxidation, with the above-described halogenating agent. Hydroxyrugulovamine is converted with this reaction into a corresponding halorugulovamine.

In this reaction, a halogenating agent is normally used at about 1 to about 50 mol, preferably about 1 to about 10 mol per mol of hydroxyrugulovamine. This reaction is normally carried out in a solvent that does not adversely affect the reaction. Example solvents include the same solvents as described for oxidation above. Preferable example solvents include the above-described halogenated hydrocarbons (e.g., chloroform, dichlormethane, carbon tetrachloride, 1,2-dichloroethane, etc.), ethers (e.g., diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, etc.,), nitriles (e.g., acetonitrile, etc.), esters (e.g., ethyl acetate, tert- butyl acetate, etc.), amides (e.g., formamide, dimethylformamide, dimethylacetoamide, etc.), ketones (e.g., acetone, methyl ethyl ketone, methyl isobutyl ketone, etc.), hydrocarbons (e.g., hexane, toluene, etc.,), aromatic arganic bases (e.g., pyridine, 2,4,6- trimethylpyridine, diazabicycloundecene, etc.) and so on; or an appropriate mixture thereof.

Although reaction temperature is not subject to limitation as long as the reaction proceeds, the reaction is normally carried out at about -20 to about 200°C, preferably about 0 to about 100°C. Reaction time is normally from about 2 minutes to about 48 hours, preferably about 10 minutes to about 6 hours, depending on reaction temperature and kind of solvent.

With respect to protection of functional groups not involved in the reaction, protective groups therefor, elimination of the protective groups etc., known groups or known means can be selected as appropriate.

Sulfonic acid esterification is carried out by reacting hydroxyrugulovamine, resulting from the above- described oxidation, with sulfonyl halides (e.g., methanesulfonyl chloride, benzenesulfonyl chloride, p- toluenesulfonyl chloride, trifluoromethanesulfonyl chloride, etc.), sulfonic anhydrides (e.g., methanesulfonic anhydride, trifluoromethanesulfonic anhydride, etc.) and so on, in the presence of an appropriate base. Hydroxyrugulovamine is converted with this reaction into a corresponding sulfonyloxyrugulovamine. It is preferable that the sulfonyloxyl group resulting from this reaction be converted into halogen atom by reacting with an appropriate inorganic halide (e.g., sodium iodide, potassium iodide, sodium bromide, potassium bromide, etc.) by a known method. In this reaction, a sulfonyl halide or sulfonic acid anhydride is used at about 1 to about 50 mol, preferably at about 1 to about 10 mol per mol of hydroxyrugulovamine. This reaction is normally carried out in a solvent that does not adversely affect the reaction. Example solvents include the same solvents other than water as described for halogenation above. It is preferable that these solvents be anhydrous. Reaction temperature and time is same as defined for halogenation above. With respect to protection of functional groups not involved in the reaction, protection groups therefor.

elimination of the protective groups etc., known groups or known means can be selected as appropriate.

Example bases include inorganic bases (e.g., sodium carbonate, potassium carbonate, etc.), organic bases (e.g., aromatic bases such as pyridine, 2,4,6-trimethylpyridine, picoline, 4-dimethylaminopyridine, 2,6-lutidine, diazabicycloundecene, etc.; tertiary amines such as triethylamine, dimethylaniline, etc.), alkali metals (e.g., sodium, potassium, etc.), alkyl lithiums (e.g., methyl lithium, butyl lithium, etc.), alkali metal amides (e.g., lithium diisopropylamide, lithium bistrimethylsilylamide, sodium bistrimethylamide, etc.), liquid ammonia and so on. Of these bases, liquid ones may be used as solvents.

In this reaction, a base is normally used at about 1 to about 50 mol, preferably about 1 to about 10 mol per mol of hydroxyrugulovamine.

Available alkyl anions include those of alkyl lithium (e.g., methyl lithium, ethyl lithium, n-butyl lithium, sec- butyl lithium, tert-butyl lithium, etc.), alkylmagnesium halides (e.g., methylmagnesium bromide, ethylmagnesium bromide, n-butylmagnesium bromide, n-pentylmagnesium bromide, etc.), alkylaluminum (e.g., trimethylaluminum, etc. ) and so on.

In this reaction, an alkyl anion etc. is normally used at about 1 to about 20 mol, preferably about 1 to about 10 mol per mol of starting compound.

This reaction is normally carried out in a solvent that does not adversely affect the reaction. Such solvents include ethers, such as tetrahydrofuran, diethyl ether, dioxane, dimethoxyethane etc.; amides, such as dimethylformamide, dimethylacetamide etc.; hydrocarbons, such as hexane, toluene, xylene, benzene etc.; aromatic organic bases, such as pyridine, 2,4,6-trimethylpyridine, diazabicycloundecene, etc.; and appropriate mixtures thereof.

Of these solvents, tetrahydrofuran, diethyl ether, dimethoxyethane, dimethylformamide, dimethylacetamide, etc. are preferable. This reaction may be carried out in the presence of appropriate inorganic salts (e.g., lithium iodide, lithium bromide, lithium chloride, etc.).

In this reaction, a salt is normally used at about 0.1 to about 50 mol, preferably about 1 to about 10 mol per mol of a starting compound.

Although reaction temperature is not subject to limitation as long as the reaction proceeds, the reaction is normally run at about -100 to 150 ^β C, preferably about -100 to 100°C, especially preferably about -70 to about 80 ^β C. Reaction time is normally from about 2 minutes to about 48 hours, preferably about 10 minutes to about 6 hours, depending on reaction temperature and kind of solvent.

With respect to protection of functional groups not involved in the reaction, protective groups therefor, elimination of the protective groups etc., known groups or known means can be selected as appropriate.

In the present invention, compounds [I], [II] and salts thereof are produced by subjecting rugulovasine, its derivative or salt thereof represented by general formula [III] to intramolecular amidation to convert the 7-lactone ring into the 5-lactam ring (process 1), followed by reduction of amide group (process 2), with hydroxyl group rearrangement and/or etherification (process 3), if necessary.

Ri5/ Ri6 and Ri7 in general formula [la] above, and Ri ₈ / Ri9 and R ₂₀ in general formula [lb] above have the same definitions as those of R ₄ , R ₅ and Re in general formula [II], respectively.

Process 1

In this process, the 7-lactone ring of rugulovasine, its derivative or salt thereof represented by general formula [III] is converted into a 5-lactam ring to give compound [la], or salt thereof. This reaction is carried out by bringing compound [III] or salt thereof into contact with a base. Available bases include alkali metal alkoxides, such as sodium methoxide, sodium ethoxide and potassium tert-butoxide, alkali metal hydroxides, such as sodium hydroxide and potassium hydroxide etc., and alkali metal hydrides, such as sodium hydride and so on. The base is used at about 1 to about 50 equivalents, preferably about 3 to about 20 equivalents per equivalent of compound [III], or salt thereof. This reaction is normally carried out in a solvent that does not adversely affect the reaction. Such solvents include alcohols, such as methanol, ethanol, sec-butanol etc., ethers, such as tetrahydrofuran, ethyl ether, dioxane, dimethoxyethane etc., and halogenated hydrocarbons, such as dichloromethane, chloroform, etc., amides, such as dimethylformamide, dimethylacetamide, etc., sulfoxides, such as dimethyl sulfoxide, etc., nitriles, such as acetonitrile, etc., hydrocarbons, such as hexane, benzene, toluene, etc., amines, such as liquid ammonia, methylamine, triethylamine, diisopropylethylamine, etc., aromatic bases, such as pyridine, 2,4,6-trimethylpyridine, picoline, 2,6- lutidine, diazabicycloundecene, etc., or an appropriate mixture thereof.

Of these solvents, methanol, ethanol, dimethylformamide, dimethylacetamide, dioxane,

dimethoxyethane, acetonitrile, etc. are preferable. It is preferable that these solvents be anhydrous.

Reaction temperature is normally about -70 to 150°C, preferably about 0 to 100°C, reaction time being from about 10 minutes to about 24 hours, preferably about 1 to about 20 hours.

Process 2

In this process, the amide group of compound [la] represented by general formula [la] is reduced to yield compound [lb] (or salt thereof) represented by general formula [lb]. This reduction reaction is carried out using a reducing agent selected from metal hydrides, such as lithium aluminum hydride, lithium borohydride, sodium cyanoborohydride or diborane. This reaction is normally carried out in a solvent that does not adversely affect the reaction. Available solvents include ethers, such as tetrahydrofuran, ethyl ether, dioxane and dimethoxyethane, etc., aromatic bases, such as pyridine, 2,4,6-trimethyl pyridine, picoline, etc., hydrocarbons such as hexane, benzene and toluene, etc.. The metal hydride is used at about 1 to about 20 equivalents, preferably about 1 to about 5 equivalents, for compound [la] or salt thereof. Reaction temperature is normally about -70 to 150°C, preferably about 0 to 100°C, reaction time being from about 5 minutes to about 30 hours, preferably about 1 hour to about 20 hours.

Process 3 [lb]

R ₂ i R ₂₂ and R ₂₃ in general formula [Ic] above have the same definitions as those of R ₄ , R ₅ and R ₆ in general formula [II], respectively, and Xi is a hydrogen atom or a hydrocarbon group. With respect to general formula [Ic], the hydrocarbon group for X_ is exemplified by the same such group as defined for general formula [I] above.

Process 3

This process achieves rearrangement of the hydroxyl group of compound [lb]. This reaction is carried out by bringing the starting compound [lb] or salt thereof into contact with an appropriate acid. Prefereble example acids include haloacetic acids (e.g., trifluoroacetic acid, etc.), inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.), organic carboxylic acids (e.g., acetic acid, citric acid, tartaric acid, oxalic acid, etc.), Lewis acids (e.g., zinc- acetic acid, boron trifluoride-ether complex, etc.) and organic sulfonic acids (e.g., benzenesulfonic acid, p- toluenesulfonic acid, camphorsulfonic acid, etc.) and so on, with especially preference given to hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, tartaric acid, p-toluenesulfonic acid, camphorsulfonic acid and so on. This reaction is normally carried out in a solvent that does not adversely affect the reaction. Such solvents include water, aqueous solutions of appropriate salts (e.g., sodium chloride, ammonium chloride, sodium dihydrogen phosphate, etc.), lower alcohols (e.g., methanol, ethanol, propanol, isopropanol, tert-butanol, hexanol, benzyl aclohols, etc.), ethers (e.g., tetrahydrofuran, ethyl ether, dioxane, dimethoxyethane, etc.), nitriles (e.g., acetonitrile, etc.), esters (e.g., ethyl formate, ethyl acetate, tert-butyl acetate, etc.), halogenated hydrocarbons (e.g., dichloromethane, chloroform, etc.), sulfoxides (e.g., dimethyl sulfoxide, etc.), amides (e.g., dimethylformamide, dimethylacetamide,

etc.); or appropriate mixtures thereof. When a solvent containing alcohols is used, an alkoxyl group corresponding to an used alcohol is formed with etherification after a rearrangement of a hydroxyl gorup. The acid is used at about 1 to about 20 equivalents, preferably about 1 to about 5 equivalents. Reaction temperature is normally about -20 to about 150°C, preferably about 0 to 50°C, reaction time being from about 10 minutes to about 7 days, preferably about 20 minutes to about 2 days. Compounds [I] and [II] or salts thereof are also produced by subjecting a compound or salt thereof of the general formula [IV] to N-substitution (process 4) above.

With respect to general formula [IV], the hydrocarbon group for Rio or X, the lower alkyl group for R_ι and the substituent that may be present on rings A and B are exemplified by the same such groups as those defined for general formula [I] above.

Compounds of general formula [IV] other than those having a hydrogen atom for X and a methyl group for Rio, ^R n (hereinafter referred to as compound [IVa]), or salt thereof are produced from setoclavine by per se known methods, e.g., the same method as for producing compound [Ilia], or salt thereof.

Process 4

In this process, a compound represented by general formula [IV], or a salt thereof is reacted with a compound having a leaving group in the presence of a base. Example bases include the same bases as described in the N-

substitution above. Of these bases, the above described alkali metal hydrides, alkali metal alkoxides, alkali metals, alkyl lithiums, alkali metal amides, etc. are preferred. Of these bases, liquid ones may be used as solvents.

Compounds having a leaving group include the same compounds as described in the N-substitution above. Of those compound, the above-described iodides, bromides, chlorides, sulfonates (e.g., p-toluenesulfonates; methanesulfonate; benzenesulfonates; trifluoromethanesulfonates, etc.), sulfates, epoxides, etc. are preferred.

This reaction is normally carried out in a solvent that does not adversely affect the reaction. Example solvents include the same solvents as described in the N- substitu ion above. Of these solvents, dimethylformamide, dimethylacetamide, tetrahydrofuran, ethyl ether, dioxane, dimethoxyethane, dimethyl sulfoxide, tert-butanol, acetonitrile benzene, toluene, pyridine, liquid ammonia, methylamine, etc. are preferred. The solvents are used singly or in combination in appropriate ratios. It is preferable that these solvents be anhydrous.

The base is used at about 1 to about 50 equivalents, preferably about 1 to about 10 equivalents, especially preferably about 1 to about 5 equivalents, for compound

[IV]. The compound having a leaving group is used at about 1 to about 40 equivalents, preferably about 1 to about 10 equivalents, especially preferably about 1 to about 8 equivalents, for compound [IV]. Reaction temperature is normally about -70 to about 150 ^β C, preferably about 0 to about 80°C, reaction time being from about 2 minutes to about 48 hours, preferably about 10 minutes to about 6 hours, especially preferably about 10 minutes to about 4 hours. A representative starting material compound for process 4 is setoclavine. Setoclavine is prepared from

agroclavine by the method of Hofmann et al. (id.).

Agroclavine has been reported as produced by Clavices purpurea by Abe et al [Annual Rep. Takeda Res. Lab., Vol.

10, p. 145 (1951)]. Agroclavine can be produced by culturing a microorganism capable of producing it in a culture medium to produce and accumulate agroclavine in the medium, and harvesting it; useful strains include variant strains capable of producing agroclavine derived by known methods, including gene manipulation, as well as Clavices o purpurea.

Compounds [I] and [II] or salts thereof are also produced by subjecting a compound [V] or a salt thereof represented by general formula [V] to oxidation (Process 5) above. 5 With respect to general formula [V], the hydrocarbon group for R_ ₂ or R ₃ . ₃ and the lower alkyl group for R ₁₄ are exemplified by the same such groups as those defined for general formula [I] above.

Compounds of general formula [V] having an optionally o substituted hydrocarbon group for R ₁₂ , or a salt thereof are produced from agroclavine by per se known methods such as process 4 above.

Compounds of general formula [V] having an optionally substituted hydrocarbon group for R ₁₃ , or salts thereof are 5 produced from N-demethylagroclavine by per se known methods such as process 4 above. N-Demethylagroclvaine has been reported as produced by ergot fungus by Yamatodani et al.

[Annual Rep. Takeda Res. Lab., Vol. 21, pp. 88-94 (1962)].

N-Demethylagroclavine has been reported as produced from 0 agroclavine by chemical conversion by E. Eich et al.

[Archiv der Pharmazie (Weinheim) , vol. 319, pp.214-218

(1985)].

Compounds of general formula [V] having alkyl group containing 2 or more carbon atoms for R ₁₄ , or salts thereof 5 are produced from elymoclavine by per se known methods such as the same method as for producing compound [Ilia], or

salt thereof. Elymoclavine has been reported as produced by Claviceps purpurea by Abe et al. [Journal of Agricultural Chemical Society of Japan, Vol. 25, p.458 (1952)].

Process 5

In this process, a compound represented by general formula [V], or a salt thereof is reacted with an oxidizing agent. Example oxidizing agents include the same oxidizing agents above. Of those oxidizing agents, selenium derivatives, peracids, chromic acids, chromates, dichromates, permanganates, perhalic acids, perhalates, peracid esters and peroxides are preferred.

In this oxidation, the oxidizing agent is used at about 1 to about 10 mol, preferably about 1 to about 3 mol per mol of a starting compound.

This reaction is normally carried out in a solvent that does not adversely affect the reaction. Example solvents include the same solvents as described in the oxidation above. Of those solvents, the above-described water, formic acid, acetic acid, ketones, amides, halogenated hydrocarbons, nitriles, aromatic organic bases, etc. are preferred.

This reaction may be carried out in the presence of an appropriate inorganic acids (e.g, sulfuric acid, phosphoric acid, etc.).

Although reaction temperature is not subject to limitation as long as the reaction proceeds, the reaction is normally carried out at about -20 to about 150°C,

preferably about 0 to about 100 ^β C. Reaction time is from about 1 minute to about 48 hours, preferably about 5 minutes to about 6 hours.

With respect to protection of functional groups not involved in the reaction, protective groups therefor, elimination of the protective groups etc., known groups or known means can be selected as appropriate.

Compound [I] or salt thereof is also produced by subjecting a compound [VI] or a salt thereof represented by the general formula:

wherein each of R24 and Y is a hydrogen atom or an optionally substituted hydrocarbon group; R25 is a lower alkyl group to N-substitution (Process 6).

Each of R24 and Y in general formula [VI] above have the same definitions as Ri in general formula [I]. R25 in general formula [VI] above have the same definitions as R3 in general formula [I].

N-Substitution is carried out by reacting the starting material with a compound having a leaving group in the presence of a base.

Process 6 [I]

Process 6

In this process, a compound represented by general formula [VI], or a salt thereof is reacted with a compound having a leaving group in the presence of a base as the same manner described in the process 4.

A representive starting compound for process 6 is N- demethylsetoclavine (6-norsetoclavine) . N- Demethylsetoclavine has been reported as produced by ergot fungus by E.Ramstad et al. [Lloydia, _, 30., 441-447 (1967)], and also has been reported as produced from N- demethylagroclavine by microbial oxidation by E.Eich et al. [Planta Medica, 282-283(1985)].

In the process 6, a starting compound of general formula [VI] is also produced from the compound of general formula [V] , wherein R ₁₃ is a hydrogen atom, by the same method as process 5.

When the desired product is obtained in a free form by the above process, it may be converted into its salt by a conventional method. When the desired product is obtained as a salt, it can be converted into a free form or another salt by a known method. The thus-obtained compound or salt thereof can be isolated and purified from the reaction mixture by known methods, such as redissolution, concentration, solvent extraction, fractional distillation, crystallization, recrystallization and chromatography.

Since the compound [I] of the present invention can be present in at least 4 isomers, because it has at least 2 asymmetric carbon atoms, at both 5- and 8-positions or at both 5- and 10-positions, these isomers and mixtures thereof are included in the scope of the present invention. Similarly, steric isomers can occur when there is an asymmetric carbon atom in a substituent thereof; these isomers and mixtures thereof are also included in the scope of the present invention.

Examples

By the following reference examples, working examples, experimental examples and formulation examples, the present invention will be described in further detail, but they are not intend to limit the invention in any manner.

Percent values are % by volume, and the mixing ratios in mixed solvents mean the volume ratio of each solvent, unless otherwise stated.

Symbols used in the specification are of the following meaning. s : singlet d : doublet dd double doublet ddd : double double doublet t triplet dt : double triplet q : quartet dq : double quartet quint : quintet dquint :: double quintet m multiplet dm double multiplet br broad

J ! coupling constant (Hz)

Reference Example 1 Production of agroclavine

One loopful of Hypomyces aurantius strain IFO 7773 grown on agar slant medium (potato dextrose-agar medium produced by Difco) was inoculated to a seed medium (500 ml, adjusted to pH 7.0) comprising 2.0% (w/v) glucose, 3.0% (w/v) maltose, 0.3% (w/v) yeast extract, 1.5% (w/v) raw soybean flour, 1.0% (w/v) corn steep liquor, 0.5% (w/v) polypeptone, 0.3% (w/v) sodium chloride in a 2-liter Sakaguchi flask, followed by 54 hours of culturing at 24°C

on a reciprocal shaker, to yield a seed culture. This seed culture broth was transferred to a main fermentation medium (120 liter, pH 6.0) containing 0.5% (w/v) glucose, 5.0% (w/v) mannitol, 0.2% (w/v) corn steep liquor, 1.0% 5 (w/v) succinic acid, 0.03% (w/v) magnesium sulfate heptahydrate and 0.1% (w/v) monopotassium phosphate in a 200-liter stainless steel tank, followed by 5 days of spinner culture at an aeration rate of 120 /minute,

/ internal pressure of 1 kg/m ² , stirring rate of 120 rpm and 0 temperature of 28°C. Subsequently, the obtained culture broth (240 liter) was adjusted to pH 3.0, and filtered in the presence of Hyflo Super Cel (produced by Johns Manvile, USA). The obtained filtrate (205 liter) was adjusted to pH 10.0 and 2 times extracted with ethyl 5 acetate (70 liter). The organic layer (120 liter) was extracted with 0.05 N hydrochloric acid (40 liter) twice. The aqueous layer was adjusted to pH 4.0 and concentrated to about 6 liter under reduced pressure. The concentrate was subjected to column chromatography with Diaion HP-20 0 (produced by Mitsubishi Kasei, Japan, 20 - 50 mesh, 1.0 liter) and washed with water (4.0 liter), followed by elution with 50% aqueous methanol (2.5 liter) and 50% methanol-0.05N aqueous hydrochloric acid (1.0 liter). The eluate was concentrated to about 0.6 liters under reduced 5 pressure and adjusted to pH 10.0, followed by extraction with ethyl acetate (250 ml) twice. The ethyl acetate layer was twice washed with water (150 ml) twice, dried over anhydrous sodium sulfate and concentrated. The concentrate was crystallized from ethyl acetate to give 0 agroclavine as white crystals (2.87 g). Elemental analysis (for C_ ₆ H_βN ₂ )

Calculated: C, 80.63; H, 7.61; N, 11.75 _' Found : C, 80.26; H, 7.67; N, 11.86 Optical rotation : [ ]D (27°C) -185° (c 0.477, in

35 pyridine)

Reference Example 2

Production of (5R, 8S)-9,10-didehydro-6,8-dimethyl-8- ergolinol (compound A ( (+)-setoclavine) )

Agroclavine (1.51 g) as obtained in Reference Example 1 was dissolved in 50% aqueous acetone (60 ml) containing 2 N sulfuric acid (3.5 ml), followed by stirring at 70°C. To this solution, an aqueous solution (60 ml) of potassium dichromate (1.87 g) heated to 70°C was added; 1 minute later, 2 N sulfuric acid (4.1 ml) was added, followed by stirring at 70 ^β C for 15 minutes. The reaction mixture was cooled to 0 ^β C and adjusted to pH 2.5, after which it was stirred at room temperature for 1 hour and filtered through filter paper. The filtrate was adjusted to pH 10.0 and extracted with chloroform-2-propanol mixtures (4:1, 200 ml) twice. The obtained organic layer was washed with a 2% (w/v) aqueous solution of sodium bicarbonate (200 ml), after which it was dried over anhydrous sodium sulfate and concentrated to dryness to give as crude powdery product (1.13 g). To this powdery product, methanol was added; the obtained precipitate was washed with methanol to give compound A as powdery product (724 mg, purity:91%). Compound A (1.07g, purity:91%) as obtained with the same method above was dissolved in dichloroethame-ethanol mixture, crystallized, and washed with ethanol to give compound A as pale yellow crystals (558mg) . Elemental analysis for (C ₁₆ H ₁₈ N ₂ O)

Calculated: C, 75.56; H, 7.13; N, 11.01 Found : C, 75.30; H, 7.19; N, 10.93 E NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 2.53 (IH, d, J=11.3 Hz), 2.57 (3H, s), 2.68 (IH, ddd, J=14.6, 11.4, 1.8 Hz), 2.81 (IH, dd, J=11.3, 1.3 Hz), 3.04 (IH, br), 3.07 (IH, ddd, J=11.4, 5.7, 1.9 Hz), 3.55 (IH, dd, J=14.6, 5.7 Hz), 6.41 (IH, brs), 6.92 (IH, t, J=1.7 Hz), 7.15-7.25 (3H, m) , 7.95 (IH, brs).

Optical rotation: [c]D (27°C) +173° (c 0.355, in pyridine) .

Reference Example 3 Production of N-cyano-N-demethylagroclavine

Agroclavine (1.89 g) as obtained in Reference Example 1 was dissolved in dichloromethane (60ml). To this solution, cyanogen bromide (2.21g) was added, followed by stirring at 25 ^β C for 2 hours. The reaction mixture was diluted with hexane (60ml) and ethyl acetate (120ml), washed with saturated aqueous solution of sodium bicarbonate (100ml) and saturated saline (100ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product. To this residue, isopropyl ether was added. The obtained precitate was washed with isopropyl ether to give N-cyano-N- demethylagroclavine as powdery product (1.51g). Elemental analysis (for C ₁₆ H _{15 3} .O.2H ₂ O)

Calculated: C, 75.98; H, 6.14; N, 16.61 Found : C, 75.95; H, 6.21; N, 16.76

^X H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.81 (3H, d, J=0.7Hz), 3.13 (IH, ddd, J=13.4, 11.6, 1.6 Hz), 3.24 (IH, ddd, J=11.6, 8.9, 3.5 Hz), 3.38 (IH, dd, J=13.4, 3.5 Hz), 3.79 (IH, brd, J=17.1 Hz), 3.83 (IH, brd, J=8.9Hz), 3.87 (IH, brd, J=17.1Hz), 6.25 (IH, brs), 6.95 (IH, t, J=1.7 Hz), 7.02 (IH, m) , 7.21 (2H, m) , 8.03 (IH, brs). IR (KBr) : 3320, 2920, 2850, 2210, 1600, 1450, 1350, 1230, 1210, 1090, 740 cm-l

Reference Example 4

Production of N-demethylagroclavine

N-Cyano-N-demethylagroclavine (0.98g) as obtained in

Reference Example 3 was dissolved in acetic acid (22ml)- water (2.5ml) mixture. To this solution, zinc dust (4.3g) was added, followed by stirring at 100°C for 6 hours. The reaction mixture was filtered through filter paper; the

filtrate was concentrated to dryness. To the obtained residue, saturated aqueous solution of sodium bicarbonate (50ml) was added, followed by extracting with ethyl acetate (60ml) twice. The obtained organic layer was washed with saturated aqueous solution of sodium hydrogen carbonate and saturated saline (each 50ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product. To this crude powdery product, ethyl ether was added. The obtained precipitate was washed with ethyl ether to give N-demethylagroclavine as powdery product (0.64g). Elemental analysis (for C ₁₆ H ₁₅ N ₂ .H ₂ O)

Calculated: C, 76.63; H, 7.37; N, 11.91 Found : C, 76.36; H, 7.40; N, 11.91 *H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.76 (3H, brs), 2.78 (IH, ddd, J=14.0, 11.1, 1.6 Hz), 2.99 (IH, ddd, J=ll.l, 9.1, 4.0 Hz), 3.08 (IH, dd, J=14.0, 4.0 Hz), 3.38 (IH, dm, J=15.7 Hz), 3.49 (IH, dm, J=15.7 Hz), 3.55 (IH, ), 6.21 (IH, brs), 6.87 (IH, t, J=1.6 Hz), 7.01 (IH, dquint, J=1.2, 4.2 Hz), 7.17 (2H, d, J=4.2 Hz), 8.01 (IH, brs) .

Reference Example 5

Production of N-demethyl-N-ethylagroclavine N-Demethylagroclavine (290mg) as obtained in

Reference Example 4 was dissolved in acetonitrile. To this solution, triethylamine (0.89ml) and ethyl iodide (0.44ml), followed by stirring at 50 ^β C for 1 hour. The reaction mixture was concentrated to dryness. To the obtained residue, ethyl acetate (30ml) was added. This solution was washed with water and saturated saline (each 50ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product. To the crude powdery product, ethyl ether was added. The obtained precipitate was washed with ethyl ether to give N-demethyl-N-ethylagroclavine as powdery product (206mg).

Elemental analysis (for C ₁₇ H ₂₀ N ₂ .O.3H ₂ O)

Calculated: C, 79.21; H, 8.05; N, 10.87 Found : C, 78.95; H, 7.91; N, 10.71 *H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.15 (3H, t, J=7.1 Hz), 1.79 (3H, brs), 2.77 (IH, dq, J=13.0, 7.1 Hz), 2.83 (IH, ddd, J=11.6, 11.6, 1.6 Hz), 2.88 (IH, m) , 3.03 (IH, dq, J=13.0, 7.1 Hz), 3.06 (IH, dm, J=15.0 Hz), 3.26 (IH, brd, J=15.0 Hz), 3.32 (IH, d, J=11.6 Hz), 3.75 (IH, m), 6.18 (IH, brs), 6.88 (IH, t, J=1.8 Hz), 7.01 (IH, m), 7.16 (2H, d, J=4.5 Hz), 7.97 (IH, brs).

Reference Example 6 Production of elymoclavine

A culture broth as obtained with the same method as described in Reference Example 1 (4150 liter) was adjusted to pH 3.0, and filtered in the presence of Hyflo Super Cel (produced by Johns Manvile, USA). The obtained filtrate (4240 liter) was adjusted to pH 11.0 and extracted with ethyl acetate (1400 liter). The organic layer (1320 liter) was extracted with 0.05N hydrochloric acid (400 liter). The water layer was adjusted to pH 4.0 and concentrated to about 150 liter under reduced pressure. The concentrate was subjected to column chromatography with Diaion HP-20 (produced by Mitsubishi Kasei, Japan, 20-50 mesh, 20 liter); the fraction eluted with water was collected and concentrated to about 4.5 liter under reduced pressure. The concentrate was subjected to column chromatography with Diaion HP-20 (50-100 mesh, 1.0 liter) and washed with water (3.0 liter), followed by elution with 10% aquecous methanol (3.0 liter). The eluate was adjusted to pH 10.0, followed by extraction with ethyl acetate (1.0 liter) twice. The ethyl acetate layer was twice washed with water (0.5 liter), dried over anhydrous sodium sulfate, and concentrated, followed by filtering through filter paper. The filtrate was concentrated to dryness to give a powdery product (4.4g). This powder

product was subjected to reverse-phase preparative HPLC [carrier, octadecylsilane (ODS), YMC-pack S-363 1-15, produced by YMC, Japan; mobile phase 14% acetonitrile / 0.05% aquecous solution of trifluoroacetic acid] in 4 times; the fraction eluted in amount of 400 to 480 ml was collected and concentrated to about 200ml. The concentrated solution was adjusted to pH 10 and three times extracted with ethyl acetate (100ml). The obtained organic layer was washed with water (100ml), dried over anhydrous sodium sulfate and concentrated. The concentrate was crystallized from ethyl acetate to give elymoclavine as white crystals (504mg). Elemental analysis (for C ₁₆ H ₁₈ N ₂ .O.2H ₂ O)

Calculated: C, 74.51; H, 7.19; N, 10.86 Found : C, 74.53; H, 7.19; N, 10.82

^X H NMR (δ ppm, in deuterochloroform-deuteromethanol (3:1), 300 MHz): 2.53 (3H, s), 2.63 (IH, ddd, J=11.5, 9.4, 3.8 Hz), 2.81 (IH, ddd, J=14.1, 11.7, 1.4 Hz), 3.03 (IH, brdt, J=16.3, 3.0 Hz), 3.36 (IH, dd, J=14.1, 4.0 Hz), 3.45 (IH, brd, J=16.3 Hz), 3.80 (IH, brd, J=8.8 Hz), 4.08 (IH, brd, J=12.9 Hz), 4.13 (IH, brd, J=12.9 Hz), 6.46 (IH, brs), 6.92 (IH, brs), 6.97 (IH, d, J=6.9 Hz), 7.13 (IH, dd, J=8.1, 6.9 Hz), 7.19 (IH, d, J=8.1 Hz).

Example 1

Production of (5R, 10S) and (5S, 10R)-8,9-didehydro-6,8- dimethyl-7-oxo-10-ergolinol (compound 1)

Compound l(one of isomers is represented) Rugulovasine (A/B mixture, 80% purity, 20 g) was dissolved in methanol (300 ml). To this solution, 28%

(w/w) sodium methoxide-methanol solution (200 ml) was added, followed by refluxing for 80 minutes. After the reaction mixture was cooled to 0°C, acetic acid (59 ml) was added dropwise over a 30-minute period. After ethyl

5 acetate (500 ml) was added, the precipitate was collected by filtration and washed with ethyl acetate (300 ml x 2), water (250 ml x 2), 50% aqueous acetone (200 ml) and ethyl ether (100 ml) to give a powder of title compound 1 (4.62 g) . The filtrate and ethyl acetate washings (about 1.3 0 liter combined) were concentrated to about 300 ml. After toluene (250 ml) was added to the concentrate, the precipitate was collected by filtration and washed with ethyl acetate (250 ml x 2), water (150 ml x 2), 50% aqueous acetone (100 ml) and ethyl ether (100 ml) to give

15 compound 1 as powdery product (4.29 g). Elemental analysis (for C ₁₆ H ₁₆ N ₂ O ₂ )

Calculated: C, 71.62; H, 6.01; N, 10.44 Found : C, 71.19; H, 5.89; N, 10.27 ¹ H NMR ( δ ppm, in dimethyl sulfoxide-d ₆ , 300 MHz): 1.89

20 (3H, d, J=1.2 Hz), 3.06 (3H, s), 3.07 (IH, dd, J=14, 12.4 Hz), 3.30 (IH, dd, J=14, 4.5 Hz), 3.72 (IH, dd, J=12.4, 4.5 Hz), 5.30 (IH, s), 7.10 (IH, t, J=7.6 Hz), 7.13 (IH, brs), 7.22 (IH, d, J=7.1 Hz), 7.26 (IH, brs), 7.28 (IH, d, J=8.1 Hz), 10.83 (IH, brs).

25

Example 2

Production of (5R, 10R) and (5S, 10S)-8,9-didehydro-6,8- dimethyl-7-oxo-10-ergolinol (compound 2)

2(one of isomers is represented)

Rugulovasine (A/B mixture, 80% purity, 25 g) was dissolved in methanol (450 ml). To this solution, 28% (w/w) sodium methoxide-methanol solution (44 ml) was added, followed by refluxing for 9 hours. After the reaction mixture was cooled to 0°C, acetic acid (12.8 ml) was added dropwise. This mixture was concentrated to about 100 ml. After ethyl acetate (300 ml) was added to the concentrate, the precipitate was collected by filtration and washed with ethyl acetate (100 ml x 2), water (80 ml x 2), 50% aqueous acetone (150 ml) and ethyl ether (150 ml) to give compound 1 as powdery product (8.10 g). The filtrate and ethyl acetate, 50% aqueous acetone and ethyl ether washings were combined and concentrated to about 300 ml. After toluene (300 ml) was added to the concentrate, the precipitate was collected by filtration and washed with a toluene-hexane mixture (3:1, 300 ml) and ethyl ether (100 ml) to give as powdery (2:5 compound 1 and 2 mixture, 14.7 g, 50% purity). The filtrate and toluene-hexane washings were combined and concentrated; this concentrate was diluted with ethyl acetate (400 ml) and washed with water (200 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give compound 2 as crude powdery product (4.8 g). The powdery product was subjected to silica gel column chromatography (200 ml, Kieselgel 60, 70-230 mesh, E.Merck, Art. 7734, Germany) and washed with acetone-toluene (2:8, 600 ml), after which the fractions eluted with acetone-toluene (25:75, 400 ml) were collected and concentrated. To the residue, ethyl acetate was added; the obtained precipitate was washed with ethyl ether to give compound 2 as powdery product (1.00 g) . Elemental analysis (for Ci ₆ Hι _{6 2} θ ₂ *0.5H ₂ θ)

Calculated: C, 69.30; H, 6.18; N, 10.10 Found : C, 68.78; H, 6.32; N, 9.59 IR NMR (δ ppm, in dimethyl sulfoxide-d ₆ , 300 MHz): 1.76 (3H, d, J=1.4 Hz), 2.62 (IH, ddd, J=15.3, 10.9, 1.3 Hz),

3.02 (3H, s), 3.38 (IH, dd, J=15.3, 5.4 Hz), 3.73 (IH, ddd, J=10.9, 5.4, 1.5 Hz), 5.77 (IH, d, J=1.0 Hz), 6.16 (IH, t, J=1.2 Hz), 7.01 (IH, brs), 7.12 (IH, d, J=4.0 Hz), 7.12 (IH, d, J=5.0 Hz), 7.20 (IH, quint, J=4.4 Hz), 10.76 (IH, brs).

Example 3

(A) Production of (5R, 10S) and (5S, 10R)-8,9-didehydro-

6,8-dimethyl-10-ergolinol (compound 3A)

HN Compound 3 A(one of isomers is represented)

The compound 1 (400 mg) as obtained in Example 1 was suspended in dioxane (13 ml). To this suspension, lithium aluminum hydride (powder, 84 mg) was added, followed by refluxing for 3 hours. After further addition of lithium aluminum hydride (powder, 28 mg), the reaction mixture was refluxed for 2 hours. The reaction mixture was cooled to 10 ^β C; an aqueous solution of 0.2 M citric acid (20 ml) was gradually added dropwise to decompose excess lithium aluminum hydride. The solution was adjusted to pH 10 and twice extracted with ethyl acetate (30 ml). The obtained organic layer was washed with a 2% (w/v) aqueous solution of sodium carbonate (30 ml), after which it was extracted with 0.1 Ν hydrochloric acid (30 ml) and water (30 ml). The obtained aqueous layers were combined and adjusted to pH 10, after which the layers were twice extracted with ethyl acetate (30 ml). The obtained organic layer was washed with saturated saline (20 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (307 mg). To this powdery product, ethyl ether was added; the precipitate was washed with

ethyl ether to give compound 3A as powdery product (88 mg).

Elemental analysis ( for C ₁₆ H ₁₈ N ₂ OΗ ₂ O)

Calculated : C , 70 . 56 ; H, 7 . 40 ; N , 10 . 29 Found : C , 70 .86 ; H, 6 .96 ; N, 10.13

*H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.78 (3H, s), 2.50 (3H, s), 2.61 (IH, dd, J=11.9, 4.6 Hz), 2.83 (IH, brd, J=16.5 Hz), 2.92 (IH, ddd, J=14.3, 11.9, 1.7 Hz), 3.09 (IH, brs), 3.17 (IH, dd, J=14.3, 4.6 Hz), 3.26 (IH, brd, J=16.5 Hz), 6.48 (IH, brs), 6.94 (IH, t, J=1.7 Hz), 7.21 (IH, dd, J=8.0, 6.9 Hz), 7.26 (IH, dd, J=8.0, 1.6 Hz), 7.29 (IH, dd, J=6.9, 0.9 Hz), 7.96 (IH, brs).

(B) Production of (5R, 10R) and (5S, 10S)-8,9-didehydro- 6,8-dimethyl-10-ergolinol (compound 3B)

is represented)

The compound 2 (5.20 g) as obtained in Example 2 was dissolved in dioxane (500 ml). To this solution, lithium aluminum hydride (powder, 1.13 g) was added, followed by refluxing for 90 minutes. The reaction mixture was cooled to 10°C; methanol (10 ml) was gradually added dropwise to decompose excess lithium aluminum hydride. The reaction mixture was diluted with a 10% (w/v) aqueous solution of citric acid (62 ml) and water (200 ml). The dilution was adjusted to pH 2 by the addition of hydrochloric acid and stirred at room temperature for 2 hours. The reaction mixture was concentrated to about 200 ml, diluted with water (250 ml), adjusted to pH 3.5 and washed with ethyl acetate (500 ml). The aqueous layer was concentrated to about 400 ml, subjected to column chromatography with

Diaion HP-20 (20 - 50 mesh, 200 ml, produced by Mitsubishi Kasei, Japan), washed with water (1000 ml) and 10% aqueous methanol (1000 ml); the fractions eluted with 30% aqueous methanol (1000 ml) were collected, adjusted to pH 7 and concentrated to about 250 ml, after which it was adjusted to pH 10 and twice extracted with ethyl acetate (200 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (1.64 g). This powdery product was subjected to silica gel column chromatography (60 ml, E.Merck, Art.

7734, Germany) and washed with acetone-toluene (1:9, 200 ml); the fractions eluted with acetone-toluene (15:85, 300 ml) were collected and concentrated. To the obtained residue, ethyl ether was added; the precipitate was washed with ethyl ether to give compound 3B as powdery product (408 mg) . Elemental analysis (for C ₁₆ H ₁₈ N ₂ OO.2H ₂ O)

Calculated: C, 74.50; H, 7.19; N, 10.86 Found : C, 74.60; H, 7.22; N, 10.52 ^X H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.65 (3H, s), 2.59 (3H, s), 2.71 (IH, dd, J=15.1, 11.0 Hz), 2.96 (IH, brd, J=17.6 Hz), 3.01 (IH, dd, J=15.1, 4.9 Hz), 3.11 (IH, brd, J=17.6 Hz), 3.20 (IH, dd, J=11.0, 4.9 Hz), 5.66 (IH, brs), 6.86 (IH, t, J=1.4 Hz), 7.22 (IH, dd, J=7.9, 1.5 Hz), 7.25 (IH, dd, J=7.9, 6.7 Hz), 7.31 (IH, dd, J=6.7, 1.5 Hz), 7.96 (IH, brs).

Example 4

Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-6,8- dimethyl-8-ergolinol (compound 4)

The compound 1 (3.54 g) as obtained in Example 1 was dissolved in tetrahydrofuran (200 ml). After this solution was cooled to 0°C, lithium aluminum hydride (powder, 1.00 g) was added, followed by stirring at room temperature for 3 hours. The reaction mixture was gradually poured over ice (600 g) to decompose excess

lithium aluminum hydride. After the reaction mixture was filtered, the obtained precipitate containing compound 4 was suspended in 0.1 N hydrochloric acid (500 ml); this suspension was stirred at room temperature for 2 hours and filtered. The filtrate was adjusted to pH 4 and kept standing at 4°C for 18 hours. This filtrate was concentrated to about 250 ml, subjected to column chromatography with Diaion HP-20 (20 - 50 mesh, 40 ml, produced by Mitsubishi Kasei, Japan), washed with water (200 ml); the fractions eluted with 20% aqueous methanol (200 ml) and 5 mM hydrochloric acid/20% aqueous methanol (200 ml) were collected. These fractions were combined, adjusted to pH 7 and concentrated to about 50 ml. The aqueous layer was adjusted to pH 10.5 and extracted with ethyl acetate (35 ml) twice. The organic layer was dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (410 mg). To this crude powdery product, ethyl ether was added; the precipitate was collected by filtration and washed with ethyl ether to give compound 4 as powdery product (335 mg). Elemental analysis (for C ₁₆ H ₁₈ N ₂ OO.2H ₂ O)

Calculated: C, 74.50; H, 7.19; N, 10.86 Found : C, 74.64; H, 6.93; N, 10.93 *H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 2.53 (IH, d, J=11.3 Hz), 2.57 (3H, s), 2.68 (IH, ddd, J=14.6, 11.4, 1.7 Hz), 2.81 (IH, dd, J=11.3, 1.3 Hz), 3.06 (IH, ddd, J=11.4, 5.7, 1.9 Hz), 3.07 (IH, brs), 3.54 (IH, dd, J=14.6, 5.7 Hz), 6.41 (IH, brs), 6.92 (IH, t, J=1.7 Hz), 7.15-7.25 (3H, m) , 7.97 (IH, brs).

Example 5

(A) Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-

6,8-dimethyl-8-methoxyergoline (compound 5)

The compound 3A (171 mg) as obtained in Example 3 was dissolved in methanol (4.6 ml). To this solution, camphorsulfonic acid (280 mg) was added, followed by

stirring at room temperature for 13 hours. The reaction mixture was diluted with 10 mM hydrochloric acid (15 ml) and washed with ethyl acetate (15 ml). The aqueous layer was adjusted to pH 9 and extracted with ethyl acetate (10 ml) twice. The obtained organic layer was washed with a saturated aqueous solution of sodium hydrogen carbonate (10 ml) and saturated saline (10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (162 mg). To this powdery product, ethyl acetate and ethyl ether were added; the precipitate was washed with ethyl ether to give compound 5 as powdery product (104 mg) . Elemental analysis (for C ₁₇ H ₂₀ N ₂ OO.3H ₂ O)

Calculated: C, 74.58; H, 7.47; N, 10.23 Found : C, 74.56; H, 7.46; N, 10.12

¹ H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.33 (3H, s), 2.43 (IH, d, J=12.2 Hz), 2.53 (3H, s), 2.79 (IH, ddd, J=14.6, 11.5, 1.5 Hz), 3.01 (IH, dd, J=11.5, 5.6 Hz), 3.02 (IH, d, J=12.2 Hz), 3.40 (3H, s), 3.54 (IH, dd, J=14.6, 5.6 Hz), 6.39 (IH, brs), 6.94 (IH, t, J=1.7 Hz), 7.19-7.25 (3H, m), 7.95 (IH, brs).

(B) Production of (5R, 8S) and (5S, 8R)-9,10-didehydro- 6,8-dimethyl-8-methoxyergoline (compound 5) The compound 4 (33 mg) as obtained in Example 4 was dissolved in methanol (2.0 ml). To this solution, camphorsulfonic acid (36 mg) was added, followed by stirring at room temperature for 17 hours. The reaction mixture was diluted with 10 mM hydrochloric acid (10 ml) and washed with ethyl acetate (10 ml). The aqueous layer was adjusted to pH 9 and extracted with ethyl acetate (10 ml) twice. The obtained organic layer was washed with a saturated aqueous solution of sodium hydrogen carbonate (10 ml) and saturated saline (10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give compound 5 as powdery product (29 mg).

Example 6

Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-6,8- dimethyl-8-ethoxyergoline (compound 6) The compound 3A (230 mg) as obtained in Example 3 was dissolved in ethanol (8.5 ml). To this solution, camphorsulfonic acid (436 mg) was added, followed by stirring at room temperature for 80 minutes. The reaction mixture was diluted with 2% (w/v) aqueous solution of sodium carbonate (5 ml) and concentrated to about 5 ml. After water (15 ml) was added, the obtained residue was adjusted to pH 8.5 and extracted with ethyl acetate (15 ml) twice. The obtained organic layer was washed with a saturated aqueous solution of sodium hydrogen carbonate (15 ml) and saturated saline (15 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (220 mg). To this powdery product, acetonitrile was added; the precipitate was washed with acetonitrile to give compound 6 as powdery product (116 mg) .

Elemental analysis (for C ₁₈ H ₂₂ N ₂ OO.3H ₂ O)

Calculated: C, 75.12; H, 7.91; N, 9.73 Found : C, 75.12; H, 7.35; N, 9.73 ^X H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.14 (3H, t, J=7.0 Hz), 1.33 (3H, s), 2.44 (IH, d, J=12.2 Hz), 2.49 (3H, s), 2.73 (IH, ddd, J=14.6, 11.4, 1.8 Hz), 2.99 (IH, ddd, J=11.4, 5.6, 1.8 Hz), 3.03 (IH, dd, J=12.2, 1.2 Hz), 3.51 (IH, dd, J=14.6, 5.6 Hz), 3.61 (IH, dq, J=9.4, 7.0 Hz), 3.79 (IH, dq, J=9.4, 7.0 Hz), 6.34 (IH, brs), 6.92 (IH, t, J=1.7 Hz), 7.15-7.26 (3H, m) , 8.02 (IH, brs).

Example 7

Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-6,8- dimethyl-8-propoxyergoline (compound 7) The compound 3A (157 mg) as obtained in Example 3 was dissolved in 1-propanol (5.0 ml). To this solution.

camphorsulfonic acid (330 mg) was added, followed by stirring at room temperature for 65 minutes. The reaction mixture was diluted with a 2% (w/v) aqueous solution of sodium carbonate (5 ml) and water (10 ml). After being adjusted to pH 8.5, the diluent was extracted with ethyl acetate (15 ml) twice. The obtained organic layer was washed with a saturated aqueous solution of sodium hydrogen carbonate (15 ml) and saturated saline (15 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (182 mg). To this powdery product, acetonitrile was added; the mixture was kept at 4°C. The precipitate was collected by filtration and washed with acetonitrile to give compound 7 as powdery product (24 mg). The mother liquor and washings were combined, concentrated to dryness and subjected to reverse-phase preparative HPLC [carrier, octadecylsilane (ODS), YMC-pack, SH-343, produced by YMC, Japan; mobile phase 40% and 42% acetonitrile/0.01 M phosphate buffer (pH 6.3, 600 ml and 400 ml)]; the fractions eluted in amounts of 700 to 860 ml were collected and concentrated to about 20 ml. The concentrated solution was adjusted to pH 9 and twice extracted with ethyl acetate (15 ml). The obtained organic layer was washed with a saturated aqueous solution of sodium hydrogen carbonate (15 ml) and saturated saline (15 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give compound 7 as powdery product (69 mg) .

Elemental analysis ( for CιgH ₂₄ N ₂ θ)

Calculated : C , 76 .99 ; H , 8.16 ; N, 9 . 45 Found : C , 76 . 51 ; H, 8 . 39 ; N , 9 . 31

^ ^• H NMR ( δ ppm, in deuterochloroform, 300 MHz): 0.85 (3H, t, J=7.4 Hz), 1.33 (3H, s), 1.53 (2H, dquint, 3=1 . 2 , 7.4 Hz), 2.43 (IH, d, J=12.2 Hz), 2.49 (3H, s), 2.74 (IH, ddd, J=14.5, 11.4, 1.7 Hz), 2.99 (IH, ddd, J=11.4, 5.6, 1.8 Hz), 3.02 (IH, dd, J=12.2, 1.2 Hz), 3.48 (IH, dt, J=9.3, 7.2 Hz), 3.51 (IH, dd, J=14.5, 5.6 Hz), 3.67 (IH, dt,

J=9 . 3 , 6 .7 Hz ) , 6 . 33 ( IH, brs ) , 6 .92 ( IH , t , J=1 .7 Hz ) , 7.16-7. 25 ( 3H, m) , 7.99 ( IH, brs ) .

Example 8 Production of (5R, 8S) and (5S, 8R)-8-allyloxy-9,10- didehydro-6,8-dimethylergoline (compound 8A) , and (5R, 8R) and (5S, 8S)-8-allyloxy-9,10-didehydro-6,8- dimethylergoline (compound 8B)

The compound 3A (170 mg) as obtained in Example 3 was dissolved in allyl alcohol (5.0 ml). To this solution, camphorsulfonic acid (357 mg) was added, followed by stirring at room temperature for 15 minutes. The reaction mixture was diluted with 2% (w/v) aqueous solution of sodium carbonate (5 ml) and water (15 ml) and extracted with ethyl acetate (15 ml) twice. The obtained organic layer was washed with a 2% (w/v) aqueous solution of sodium hydrogen carbonate (15 ml) and saturated saline (15 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (175 mg). To this powdery product, acetonitrile was added; the mixture was kept at 4°C. The precipitate was collected by filtration and washed with acetonitrile to give compound 8A as powdery product (52 mg). The mother liquor and washings were combined, concentrated to dryness and subjected to reverse-phase preparative HPLC [carrier, ODS, YMC-pack, SH-343; mobile phase 42% acetonitrile/0.01 M phosphate buffer (pH 6.3)]; the fractions, eluted in amounts of 550 to 700 ml and 700 to 810 ml respectively, were collected and each concentrated to about 20 ml. Each concentrated solution was adjusted to pH 8.5 and extracted with ethyl acetate (15 ml) twice. The obtained organic layer was washed with a 2% (w/v) aqueous solution of sodium hydrogen carbonate (15 ml) and saturated saline (15 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give compound 8A (53 mg) and compound 8B (11 mg).

Compound 8A

Elemental analysis (for C ₁₉ H ₂₂ N ₂ O)

Calculated: C, 77.52; H, 7.53; N, 9.52 Found : C, 76.94; H, 7.32; N, 10.09 ¹ H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 2.48 (IH, d, J=12.3 Hz), 2.48 (3H, s), 2.74 (IH, ddd, J=14.6, 11.4, 1.8 Hz), 2.99 (IH, ddd, J=11.4, 5.6, 1.7 Hz), 3.05 (IH, dd, J=12.3, 1.2 Hz), 3.53 (IH, dd, J=14.6,

5.6 Hz), 4.13 (IH, ddt, J=12.9, 5.6, 1.4 Hz), 4.36 (IH, ddt, J=12.9, 5.6, 1.3 Hz), 5.05 (IH, dq, J=10.3, 1.7 Hz),

5.22 (IH, dq, J=17.2, 1.7 Hz), 5.92 (IH, ddt, J=17.2, 10.3, 5.6 Hz), 6.31 (IH, brs), 6.93 (IH, t, J=1.7 Hz), 7.16-7.26 (3H, m) , 7.97 (IH, brs).

Compound 8B

^X H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.53 (3H, s), 2.55 (3H, s), 2.65 (IH, ddd, J=14.6, 11.4, 1.8 Hz), 2.72 (IH, d, J=10.8 Hz), 2.77 (IH, dd, J=10.8, 1.2 Hz), 3.12 (IH, ddd, J=11.4, 5.6, 2.2 Hz), 3.50 (IH, dd, J=14.6, 5.6 Hz), 4.11 (2H, brd, J=5.3 Hz), 5.18 (IH, dq, J=10.3,

1.7 Hz), 5.33 (IH, dq, J=17.2, 1.7 Hz), 5.98 (IH, ddt, J=17.2, 10.3, 5.3 Hz), 6.40 (IH, brs), 6.90 (IH, t, J=1.8 Hz), 7.19 (2H, m), 7.21 (IH, dd, J=7.4, 2.1 Hz), 7.96 (IH, brs) .

Example 9

Production of (5R, 8S) and (5S, 8R)-8-benzyloxy-9,10- didehydro-6,8-dimethylergoline (compound 9)

The compound 3A (217 mg) as obtained in Example 3 was dissolved in benzyl alcohol (3.6 ml). To this solution, camphorsulfonic acid (448 mg) was added, followed by stirring at room temperature for 20 minutes. The reaction mixture was diluted with a 2% (w/v) aqueous solution of sodium carbonate (10 ml) and twice extracted with ethyl acetate (15 ml). The obtained organic layer was two times washed with water (10 ml) and extracted with 0.2 N hydrochloric acid (17 ml) and water (10 ml). The obtained acidic aqueous layer was adjusted to pH 2.5 and washed with ethyl acetate (15 ml), after which it was adjusted to pH 9.5 and extracted with ethyl acetate (15 ml) twice. The obtained organic layer was washed with a 2% (w/v) aqueous solution of sodium hydrogen carbonate (15 ml x 2) and saturated saline (15 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (149 mg). This powdery product was subjected to column chromatography with silica gel (8 g); the fraction eluted with acetone-toluene (5:95) was collected and concentrated to dryness. Acetonitrile was added; the precipitate was washed with acetonitrile to give compound 9 as powdery product (29 mg). Elemental analysis (for C ₂₃ H _{2 2} θ*0.3H ₂ θ)

Calculated: C, 78.96; H, 7.09; N, 8.01

Found : C, 78.98; H, 6.93; N, 7.65 ¹ H NMR { δ ppm, in deuterochloroform, 300 MHz): 1.39 (3H, s), 2.47 (3H, s), 2.53 (IH, d, J=12.3 Hz), 2.76 (IH, ddd, J=14.7, 11.3, 1.8 Hz), 3.03 (IH, ddd, J=11.3, 5.6, 1.8 Hz), 3.12 (IH, dd, J=12.3, 1.2 Hz), 3.55 (IH, dd, J=14.7, 5.6 Hz), 4.65 (IH, d, J=12.1 Hz), 4.93 (IH, d, J=12.1 Hz), 6.38 (IH, brs), 6.94 (IH, t, J=1.8 Hz), 7.16-7.26 (6H, m) , 7.32 (2H, m), 7.93 (IH, brs).

Example 10

Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-l,6,8- trimethyl-8-ergolinol (compound 10)

The compound 4 (91 mg) as obtained in Example 4 was dissolved in dimethylformamide (hereinafter DMF) (3.0 ml). To this solution, sodium hydride (60%, oily, the same applies below; 24 mg) and then methyl iodide (22 μl ) were added, followed by stirring at room temperature for 90 minutes. The reaction mixture was diluted with water (15 ml), adjusted to pH 2.5 and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 9.0 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with a 2% (w/v) aqueous solution of sodium hydrogen carbonate, water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (97 mg). To this powdery product, diethyl ether was added; the precipitate was washed with diethyl ether to give compound 10 as powdery product (63 mg). Elemental analysis (for C ₁₇ H _{20 2} O)

Calculated: C, 76.09; H, 7.51; N, 10.44 Found : C, 75.61; H, 7.50; N, 10.26 ^X H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 2.53 (IH, d, J=11.3 Hz), 2.56 (3H, s), 2.67 (IH, ddd, J=14.6, 11.3, 1.6 Hz), 2.80 (IH, dd, J=11.3, 1.4 Hz), 3.03 (IH, br), 3.04 (IH, ddd, J=11.3, 5.8, 2.0 Hz), 3.51 (IH, dd, J=14.6, 5.8 Hz), 3.76 (3H, s), 6.40 (IH, brt, J=2 Hz), 6.76 (IH, d, J=1.5 Hz), 7.15 (IH, dd, J=9.2, 3.5 Hz), 7.16 (IH, t, J=9.3 Hz), 7.19 (IH, dd, J=9.4, 5.1 Hz).

Example 11

Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-8- methoxy-l,6,8-trimethylergoline (compound 11)

The mother liquor and washings obtained when the precipitate was obtained in Example 10 were concentrated to dryness and subjected to column chromatography with silica

gel (5 g, Art. 7734); the fraction eluted with acetone- toluene (15:85) was collected and concentrated to dryness to give compound 11 as powdery product (18 mg). ^X H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.32 (3H, s), 2.42 (IH, d, J=12.0 Hz), 2.51 (3H, s), 2.77 (IH, ddd, J=14.4, 11.5, 1.2 Hz), 2.98 (IH, ddd, J=11.0, 5.6, 1.5 Hz), 3.01 (IH, dd, J=12.1, 1.0 Hz), 3.39 (3H, s), 3.50 (IH, dd, J=14.4, 5.6 Hz), 3.76 (3H, s), 6.38 (IH, brs), 6.76 (IH, d, J=1.4 Hz), 7.17 (IH, dd, J=6.9, 2.3 Hz), 7.19 (IH, s), 7.20

10 (IH, d, J=7.2 Hz) . Mass analysis (EIMS): M ⁺ = 282

Example 12

Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-6,8- -c dimethyl-l-ethyl-8-ergolinol (compound 12)

The compound 4 (99 mg) as obtained in Example 4 was dissolved in DMF (3.0 ml). To this solution, sodium hydride (29 mg) and then ethyl iodide (31 μl ) were added, followed by stirring at room temperature for 70 minutes.

20 The reaction mixture was diluted with 0.15 N hydrochloric acid (12 ml), adjusted to pH 2.1 and washed with diethyl ether (10 ml) twice. The aqeous layer was adjusted to pH 9.0 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with a 2% (w/v) aqueous

25 solution of sodium hydrogen carbonate, water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (108 mg). To this powdery product, diethyl ether was added; the precipitate was washed with diethyl ether to

30 give compound 12 as powdery product (55 mg). Elemental analysis (for C ₁₈ H ₂₂ N ₂ O)

Calculated: C, 76.56; H, 7.85; N, 9.92 Found : C, 76.27; H, 7.69; N, 9.87 ¹ H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H,

35 s), 1.44 (3H, t, J=7.3 Hz), 2.52 (IH, d, J=11.3 Hz), 2.56 (3H, s), 2.67 (IH, ddd, J=14.5, 11.3, 1.7 Hz), 2.80 (IH,

dd, J=11.3, 1.4 Hz), 3.03 (IH, br), 3.06 (IH, ddd, J=11.3, 5.7, 2.0 Hz), 3.52 (IH, dd, J=14.5, 5.7 Hz), 4.13 (2H, q, J=7.3 Hz), 6.40 (IH, brt, J=2 Hz), 6.82 (IH, d, J=1.5 Hz), 7.15 (IH, dd, J=5.7, 2.9 Hz), 7.17 (IH, s), 7.17 (IH, dd, J=5.9, 3.1 Hz).

Example 13

Production of (5R, 8S) and (5S, 8R)-l-allyl-9,10-didehydro-

6,8-dimethyl-8-ergolinol (compound 13)

The compound 4 (94 mg) as obtained in Example 4 was dissolved in DMF (3.0 ml). To this solution, sodium hydride (28 mg) and then allyl bromide (33 μl ) were added, followed by stirring at room temperature for 100 minutes. The reaction mixture was diluted with 0.05 N hydrochloric acid (14 ml), adjusted to pH 2.0 and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 8.5 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with a 2% (w/v) aqueous solution of sodium hydrogen carbonate, water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (112 mg). To this powdery product, diethyl ether-hexane mixture was added; the precipitate was washed with hexane to give compound 12 as powdery product (24 mg). Elemental analysis (for C ₁₉ H ₂₂ N ₂ O)

Calculated: C, 77.52; H, 7.53; N, 9.52

Found : C, 77.13; H, 7.53; N, 9.48 ^λ E NMR (δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 2.53 (IH, d, J=ll.3 Hz), 2.56 (3H, s), 2.68 (IH, ddd, J=14.6, 11.3, 1.7 Hz), 2.80 (IH, dd, J=11.3, 1.4 Hz), 3.03 (IH, br), 3.06 (IH, ddd, J=11.3, 5.7, 1.9 Hz), 3.52 (IH, dd, J=14.6, 5.7 Hz), 4.68 (2H, dt, J=5.5, 1.6 Hz), 5.11 (IH, dq, J=17.1, 1.3 Hz), 5.19 (IH, dq, J=10.2, 1.3 Hz), 5.98 (IH, ddt, J=17.1, 10.2, 5.5 Hz), 6.40 (IH, brt, J=1.4 Hz), 6.80 (IH, d, J=1.5 Hz), 7.16 (3H, m) .

Example 14

Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-6,8- dimethyl-l-hexyl-8-ergolinol (compound 14)

The compound 4 (118 mg) as obtained in Example 4 was dissolved in DMF (3.0 ml). To this solution, sodium hydride (37 mg) and then hexyl iodide (75 μl ) were added, followed by stirring at room temperature for 50 minutes. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml), adjusted to pH 2.0 and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 9.5 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (112 mg). To this powdery product, hexane was added; the obtained precipitate was washed with hexane to give compound 14 as powdery product (104 mg). Elemental analysis (for C ₂₂ H ₃₀ N ₂ O)

Calculated: C, 78.06; H, 8.93; N, 8.28

Found : C, 77.97; H, 9.09; N, 8.26 ¹ H NMR ( δ ppm, in deuterochloroform, 300 MHz): 0.87 (3H, t, J=6.9 Hz), 1.30 (4H, brs), 1.32 (2H, m) , 1.36 (3H, s), 1.80 (2H, quint, J=7.1 Hz), 2.52 (IH, d, J=11.3 Hz), 2.56 (3H, s), 2.67 (IH, ddd, J=14.6, 11.3, 1.6 Hz), 2.80 (IH, dd, J=11.3, 1.4 Hz), 3.03 (IH, br), 3.06 (IH, ddd, J=11.3, 5.8, 1.9 Hz), 3.52 (IH, dd, J=14.6, 5.8 Hz), 4.05 (2H, t, J=7.2 Hz), 6.40 (IH, brt, J=1.5 Hz), 6.80 (IH, d, J=1.5 Hz), 7.15 (3H, m).

Example 15

Production of (5R, 8S) and (5S, 8R)-9,10-didehydro-6,8- dimethyl-l-isopropyl-8-ergolinol (compound 15)

The compound 4 (115 mg) as obtained in Example 4 was dissolved in DMF (3.0 ml). To this solution, sodium hydride (36 mg) and then isopropyl iodide (47 μl ) were added, followed by stirring at room temperature for 50

minutes. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 9.5 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (125 mg). This powdery product was subjected to column chromatography with silica gel (5 g); the fractions eluted with acetone-toluene (15:85) were collected and concentrated to dryness to give compound 15 as powdery product (42 mg) . Elemental analysis (for C ₁₉ H ₂₄ N ₂ O)

Calculated: C, 76.99; H, 8.16; N, 9.45

Found : C, 76.69; H, 8.20; N, 9.45 ^X H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 1.51 (3H, d, J=6.7 Hz), 1.52 (3H, d, J=6.7 Hz), 2.53 (IH, d, J=11.4 Hz), 2.57 (3H, s), 2.68 (IH, ddd, J=14.6, 11.3, 1.5 Hz), 2.81 (IH, dd, J=11.4, 1.2 Hz), 3.03 (IH, br), 3.07 (IH, ddd, J=11.3, 5.7, 1.8 Hz), 3.53 (IH, dd, J=14.6, 5.7 Hz), 4.61 (IH, quint, J=6.7 Hz), 6.40 (IH, brs), 6.93 (IH, d, J=1.3 Hz), 7.15 (IH, dd, J=6.1, 2.8 Hz), 7.18 (IH, dd, J=7.1, 6.1 Hz), 7.20 (IH, dd, J=7.1, 2.8 Hz).

Example 16

Production of (5R,8S)-l-benzyl-9,10-didehydro-6,8-dimethyl-

8-ergolinol (compound 16)

The compound A (113 mg) as obtained in Reference Example 2 was dissolved in DMF (2.5 ml). To this solution, sodium hydride (30 mg) and then benzyl bromide (50 μl ) were added, followed by stirring at room temperature for 40 minutes. The reaction mixture was diluted with 0.08 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 10 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with water and saturated saline

(each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (153 mg). To this powdery product, diethyl ether-hexane mixture was added; the precipitate was washed with hexane to give compound 16 as powdery product (105 mg). Elemental analysis (for C ₂₃ H ₂₄ N ₂ O)

Calculated: C, 80.20; H, 7.02; N, 8.13 Found : C, 79.79; H, 7.10; N, 8.15 ^X H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.37 (3H, s), 2.54 (IH, d, J=11.4 Hz), 2.57 (3H, s), 2.69 (IH, ddd, J=14.5, 11.3, 1.6 Hz), 2.81 (IH, dd, J=11.4, 1.3 Hz), 3.06 (IH, br), 3.08 (IH, ddd, J=11.2, 5.7, 1.8 Hz), 3.53 (IH, dd, J=14.6, 5.7 Hz), 5.27 (2H, s), 6.40 (IH, brs), 6.83 (IH, d, J=1.4 Hz), 7.09-7.18 (6H, m) , 7.25-7.31 (2H, m) .

Example 17

Production of (5R,8S)-9,10-didehydro-6,8-dimethyl-l-ethyl-

8-ergolinol (compound 17)

The compound A (293 mg) as obtained in Reference Example 2 was dissolved in DMF (6.5 ml). To this solution, sodium hydride (81 mg) and then ethyl iodide (87 μl ) were added, followed by stirring at room temperature for 40 minutes. The reaction mixture was diluted with 0.15 N hydrochloric acid (10 ml) and washed with diethyl ether (12 ml) twice. The aqueous layer was adjusted to pH 9.5 and extracted with ethyl acetate (15 ml) twice. The obtained organic layer was washed with water and saturated saline (each 15 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (318 mg). To this powdery product, diethyl ether was added; the precipitate was washed with diethyl ether to give compound 17 as powdery product (166 mg). Elemental analysis (for C ₁₈ H _{22 2} O)

Calculated: C, 76.56; H, 7.85; N, 9.92 Found : C, 76.52; H, 7.70; N, 10.01

Example 18

Production of (5R,8S)-l-carbamoylmethyl-9,10-didehydro-6,8- dimethyl-8-ergolinol (compound 18)

The compound A (125 mg) as obtained in Reference Example 2 was dissolved in DMF (2.5 ml). To this solution, sodium hydride (36 mg) and then iodoacetamide (96 mg) were added, followed by stirring at room temperature for 80 minutes. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml) and twice washed with diethyl ether (10 ml). The aqueous layer was adjusted to pH 9.5 and 4 times extracted with ethyl acetate-2-propanol (4:1, 8 ml). The obtained organic layer was washed with 5% (w/v) saline and saturated saline (each 15 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product. This powdery product was subjected to column chromatography with silica gel (5 g, E. Merck, Art. 7734, Germany); the fractions eluted with acetone:toluene (50:50)-methanol:acetone:toluene mixture (5:50:45) were collected and concentrated to dryness. To the obtained residue, diethyl ether was added; the precipitate was washed with diethyl ether to give compound 18 as powdery product (74 mg). Elemental analysis (for C ₁₈ H ₂₁ N ₃ O ₂ O.2H ₂ O)

Calculated: C, 68.64; H, 6.85; N, 13.34 Found : C, 68.65; H, 6.69; N, 13.26 ¹ H NMR (δ ppm, in deuterochloroform- methanol-d ₄ (10:1), 300 MHz): 1.33 (3H, s), 2.52 (IH, d, J=11.6 Hz), 2.55 (3H, s), 2.65 (IH, ddd, J=14.8, 11.3, 1.7 Hz), 2.83 (IH, dd, J=11.6, 1.3 Hz), 3.03 (IH, ddd, J=11.3, 5.8, 2.0 Hz), 3.51 (IH, dd, J=14.8, 5.8 Hz), 4.71 (2H, s), 6.40 (IH, brs), 6.78 (IH, d, J=1.5 Hz), 7.12 (IH, dd, J=6.6, 2.1 Hz), 7.21 (2H, m) .

Example 19

Production of (5R,8S)-l-carboxymethyl-9,10-didehydro-6,8- dimethyl-8-ergolinol (compound 19)

The compound A (102 mg) as obtained in Reference Example 2 was dissolved in DMF (2.5 ml). To this solution, sodium hydride (29 mg) and then sodium iodoacetate (89 mg) were added, followed by stirring at room temperature for 60 minutes. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 10 and 2 times washed with ethyl acetate (10 ml). The obtained water layer was adjusted to pH 7.0 and subjected to column chromatography with Diaion HP-20 (20 - 50 mesh, 5 ml

10 produced by Mitsubishi Kasei, Japan); the fractions eluted with 20% aqueous methanol were collected and concentrated to dryness. To the obtained residue, ethyl acetate was added; the precipitate was washed with ethyl acetate to -_ give compound 19 as powdery product (20 mg). Elemental analysis (for C ₁₈ H ₂₀ N ₂ O ₃ -2.3^0)

Calculated: C, 61.11; H, 7.01; N, 7.92 Found : C, 61.03; H, 6.50; N, 7.86 ^λ E NMR ( δ ppm, in methanol-d ₄ , 300 MHz): 1.33 (3H, s), 2.60 (IH, d, J=12.0 Hz), 2.60 (3H, s), 2.72 (IH, ddd,

20 J=14.5, 11.4, 1.4 Hz), 2.97 (IH, dd, J=12.0, 1.0 Hz), 3.13 (IH, ddd, J=11.4, 5.8, 1.8 Hz), 3.52 (IH, dd, J=14.5, 5.8 Hz), 4.61 (2H, s), 6.36 (IH, brs), 6.88 (IH, d, J=l.l Hz), 7.08-7.16 (3H, m) .

25

Example 20

Production of (5R,8S)-9,10-didehydro-6,8-dimethyl-l-propyl-

8-ergolinol (compound 20)

The compound A (285 mg) as obtained in Reference Example 2 was dissolved in DMF (5.0 ml). To this solution,

30 sodium hydride (82 mg) and then propyl iodide (113 μl ) were added, followed by stirring at room temperature for 45 minutes. The reaction mixture was diluted with 0.2 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 9.0 and

35 extracted with ethyl acetate (12 ml) twice. The obtained

organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (338 mg). This powdery product was subjected to column chromatography with silica gel (5 g, E. Merck, Art, 7734, Germany); the fractions eluted with acetone-toluene (20:80) were collected and concentrated to dryness. The obtained residue was dissolved in ethyl acetate-hexane (1:19, 20 ml) and the insoluble substances were removed. The residual solution was concentrated to dryness. Hexane was added; the precipitate was washed with hexane to give compound 20 as powdery product (147 mg). Elemental analysis (for C ₁₉ H ₂₄ N ₂ O)

Calculated: C, 76.99; H, 8.16; N, 9.45 Found : C, 76.66; H, 8.12; N, 9.36 ^X H NMR ( δ ppm, in deuterochloroform, 300 MHz): 0.92 (3H, t, J=7.4 Hz), 1.36 (3H, s), 1.85 (2H, tq, J=7.0, 7.4 Hz), 2.53 (IH, d, J=11.3 Hz), 2.57 (3H, s), 2.68 (IH, ddd, J=14.6, 11.3, 1.6 Hz), 2.81 (IH, dd, J=11.3, 1.2 Hz), 3.03 (IH, br), 3.06 (IH, ddd, J=11.3, 5.8, 1.9 Hz), 3.52 (IH, dd, J=14.6, 5.8 Hz), 4.03 (2H, t, J=7.0 Hz), 6.40 (IH, brt, J=l Hz), 6.81 (IH, d, J=1.4 Hz), 7.16 (3H, m) .

Example 21

Production of (5R,8S)-9,10-didehydro-6,8-dimethyl-l- methoxymethyl-8-ergolinol (compound 21)

The compound A (105 mg) as obtained in Reference Example 2 was dissolved in DMF (2.5 ml). To this solution, sodium hydride (29 mg) and then methoxymethyl chloride ether (34 μl ) were added, followed by stirring at room temperature for 30 minutes. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml), adjusted to pH 2.4 and 2 times washed with diethyl ether (10 ml). The aqueous layer was adjusted to pH 10 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml),

dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (121 mg) . To this powdery product, diethyl ether was added; the precipitate was washed with diethyl ether to give compound 21 as powdery product (59 mg). Elemental analysis (for C ₁ 8H ₂ 2N2O2)

Calculated: C, 72.46; H, 7.43; N, 9.39 Found : C, 72.20; H, 7.36; N, 9.33 ^X H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 2.53 (IH, d, J=11.3 Hz), 2.57 (3H, s), 2.67 (IH, ddd, J=14.7, 11.4, 1.7 Hz), 2.81 (IH, dd, J=11.3, 1.2 Hz), 3.03 (IH, br), 3.06 (IH, ddd, J=11.4, 5.7, 1.9 Hz), 3.24 (3H, S), 3.52 (IH, dd, J=14.7, 5.7 Hz), 5.41 (2H, s), 6.41 (IH, brs), 6.89 (IH, d, J=1.5 Hz), 7.21 (2H, m) , 7.29 (IH, dd, J=6.2, 2.9 Hz) .

Example 22

Production of (5R,8S)-9,10-didehydro-6,8-dimethyl-l-(2- methoxyethoxy)methyl-8-ergolinol (compound 22A) and (5R,8S)-9,10-didehydro-6,8-dimethyl-l-(2- methoxyethoxy)methyl-8-ergolinol maleate (compound 22B)

The compound A (119 mg) as obtained in Reference Example 2 was dissolved in DMF (2.0 ml). To this solution, sodium hydride (34 mg) and then methoxyethoxymethyl chloride (53 μl ) were added, followed by stirring at room temperature for 20 minutes. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 9.5 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (155 mg). This powdery product was subjected to column chromatography with silica gel (5 g, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-toluene mixture (20:80 - 30:70) were collected and

concentrated to dryness. The obtained residue was purified by reverse-phase preparative HPLC [carrier, ODS, YMC-pack, D-ODS-5; mobile phase 20% acetonitrile/0.05% trifluoroacetic acid]; desired fractions were collected and concentrated to about 20 ml. The concentrated solution was adjusted to pH 10 and extracted with ethyl acetate (10 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give compound 22A as powdery product (87 mg). The compound 22A was dissolved in methanol (5 ml), mixed with maleic acid (29 mg) and concentrated to dryness to give compound 22B as powdery product (119 mg). Compound 22A

¹ H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 2.53 (IH, d, J=11.4 Hz), 2.57 (3H, s), 2.66 (IH, ddd, J=14.7, 11.4, 1.8 Hz), 2.81 (IH, dd, J=11.4, 1.3 Hz), 3.03 (IH, br), 3.04 (IH, ddd, J=11.4, 5.7, 1.9 Hz), 3.35 (3H, s), 3.48 (4H, m), 3.52 (IH, dd, J=14.7, 5.7 Hz), 5.52 (2H, s), 6.41 (IH, brs), 6.91 (IH, d, J=1.5 Hz), 7.21 (2H, m) , 7.29 (IH, dd, J=6.1, 2.9 Hz). Compound 22B Elemental analysis (for C ₂ oH ₂₆ N ₂ θ ₃ -C ₄ H ₄ θ «H ₂ θ)

Calculated: C, 60.49; H, 6.77; N, 5.88

Found : C, 60.64; H, 6.64; N, 6.20

Example 23

Production of (5R,8S)-l-butyl-9,10-didehydro-6,8-dimethyl-

8-ergolinol (compound 23)

The compound A (104 mg) as obtained in Reference Example 2 was dissolved in DMF (1.5 ml). To this solution, sodium hydride (30 mg) and then butyl iodide (42 μl ) were added, followed by stirring at room temperature for 50 minutes. The reaction mixture was diluted with 0.09 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 9.5 and

extracted with ethyl acetate (10 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (108 mg). This powdery product was subjected to column chromatography with silica gel (5 g, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-toluene (15:85 - 20:80) were collected and concentrated to dryness. To the obtained residue, hexane was added; the precipitate was washed with hexane under ice cooling conditions to give compound 23 as powdery product (34 mg). Elemental analysis (for C ₂₀ H ₂₆ N ₂ O)

Calculated: C, 77.38; H, 8.44; N, 9.02

Found : C, 77.14; H, 8.31; N, 9.00 ^X H NMR (<, ppm, in deuterochloroform, 300 MHz): 0.93 (3H, t, J=7.4 Hz), 1.32 (2H, q, J=7.4 Hz), 1.36 (3H, s), 1.79 (2H, quint, J=7.3 Hz), 2.53 (IH, d, J=11.4 Hz), 2.56 (3H, S), 2.67 (IH, ddd, J=14.6, 11.2, 1.6 Hz), 2.81 (IH, dd, J=11.4, 1.4 Hz), 3.03 (IH, br), 3.06 (IH, ddd, J=11.2, 5.8, 2.0 Hz), 3.52 (IH, dd, J=14.6, 5.8 Hz), 4.06 (2H, t, J=7.0 Hz), 6.40 (IH, brt, J=l Hz), 6.80 (IH, d, J=1.5 Hz), 7.16 (3H, m) .

Example 24

Production of (5R,8S)-9,10-didehydro-6,8-dimethyl-l-(l- fluoro-2-iodoethenyl)-8-ergolinol (compound 24)

The compound A (133 mg) as obtained in Reference Example 2 was dissolved in DMF (2.0 ml). To this solution, sodium hydride (38 mg) and then 2-iodo-l,l,l- trifluoroethane (52 μl ) were added, followed by stirring at room temperature for 90 minutes. The reaction mixture was diluted with 0.15 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 10 and extracted with ethyl acetate (10 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous

sodium sulfate and concentrated to dryness to give a crude powdery product (196 mg). This powdery product was subjected to column chromatography with silica gel (5 g, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-toluene (15:85) were collected and concentrated to dryness. To the obtained residue, hexane was added; the precipitate was washed with hexane to give compound 24 as powdery product (63 mg, E/Z mixture). Elemental analysis (for C ₁₈ H ₁₈ N ₂ OFI)

Calculated: C, 50.96; H, 4.28; N, 6.60; F, 4.48; I, 29.91

Found : C, 50.88; H, 4.28; N, 6.35; F, 4.46; I, 29.58 ¹ H NMR (<5" ppm, in deuterochloroform, 300 MHz, major product = E-isomer): 1.36 (3H, s), 2.53 (IH, d, J=11.4 Hz), 2.57 (3H, s), 2.66 (IH, ddt, J=15.0, 11.5, 2.0 Hz), 2.82 (IH, dd, J=11.4, 1.4 Hz), 3.04 (IH, br), 3.07 (IH, ddd, J=11.5, 5.7, 1.9 Hz), 3.53 (IH, dd, J=15.0, 5.7 Hz), 5.97 (IH, d, J=7.2 Hz), 6.42 (IH, brs), 7.06 (IH, d, J=1.8 Hz), 7.24- 7.30 (3H, m).

Example 25

Production of (5R,8S)-l-tert-butoxycarbonylmethyl-9,10- didehydro-6,8-dimethyl-8-ergolinol (compound 25)

The compound A (113 mg) as obtained in Reference Example 2 was dissolved in DMF (2.0 ml). To this solution, sodium hydride (40 mg) and then tert-butyl iodoacetate (118 μl ) were added, followed by stirring at room temperature for 30 minutes. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The water layer was adjusted to pH 9.0 and extracted with ethyl acetate (10 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (158 mg). This powdery product was subjected to column

chromatography with silica gel (5 g, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-toluene (15:85 - 25:75) were collected and concentrated to dryness to give compound 25 as powdery product (100 mg). Elemental analysis (for C ₂₂ H ₂₈ ^N ₂ θ ₃ )

Calculated: C, 71.71; H, 7.66; N, 7.60

Found : C, 71.19; H, 7.56; N, 7.10 ^λ E NMR (δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 1.45 (9H, s), 2.52 (IH, d, J=11.3 Hz), 2.56 (3H, s), 2.68 (IH, ddd, J=14.6, 11.3, 1.7 Hz), 2.80 (IH, dd, J=11.3, 1.3 Hz), 3.04 (IH, br) , 3.07 (IH, ddd, J=11.3, 5.7, 1.9 Hz), 3.52 (IH, dd, J=14.6, 5.7 Hz), 4.69 (2H, s), 6.40 (IH, brt, J=1.4 Hz), 6.80 (IH, d, J=1.5 Hz), 7.09 (IH, dd, J=6.9, 1.8 Hz), 7.18 (2H, m) .

Example 26

Production of (5R,8S)-9,10-didehydro-6,8-dimethyl-l-pentyl-

8-ergolinol (compound 26)

A powder of compound A (101 mg) as obtained in Reference Example 2 was dissolved in DMF (2.0 ml). To this solution, sodium hydride (29 mg) and then pentyl iodide (42 μl ) were added, followed by stirring at room temperature for 40 minutes. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 9.5 and extracted with ethyl acetate (10 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (125 mg). Hexane was added to this powdery product; the insoluble substances were removed; the soluble portion was concentrated to dryness. To the obtained residue, hexane was added; the precipitate was washed with hexane under ice cooling conditions to give compound 26 as powdery product (74 mg). Elemental analysis (for C ₂₁ H ₂₈ N ₂ O)

Calculated: C, 77.74; H, 8.70; N, 8.63

Found : C, 77.50; H, 8.68; N, 8.74 ^ ^• H NMR ( δ ppm, in deuterochloroform, 300 MHz): 0.88 (3H, t, J=7.0 Hz), 1.26-1.34 (4H, m) , 1.36 (3H, s), 1.82 (2H, quint, J=7.3 Hz), 2.53 (IH, d, J=11.3 Hz), 2.57 (3H, s), 2.68 (IH, ddd, J=14.6, 11.3, 1.6 Hz), 2.80 (IH, dd, J=11.3,

1.4 Hz), 3.01 (IH, brs), 3.06 (IH, ddd, J=11.3, 5.7, 1.9 Hz), 3.52 (IH, dd, J=14.6, 5.7 Hz), 4.06 (2H, t, J=7.1 Hz), 6.40 (IH, brt, J=l Hz), 6.80 (IH, d, J=1.5 Hz), 7.16 (3H, m) .

Example 27

Production of (5R,8S)-9,10-didehydro-6,8-dimethyl-l-(2- hydroxypropyl)-8-ergolinol (compound 27)

The compound A (140 mg) as obtained in Reference Example 2 was dissolved in DMF (1.5 ml). To this solution, sodium hydride (57 mg) and then propylene oxide (40 μl ) were added, followed by stirring at room temperature for

2.5 hours. The reaction mixture was diluted with 0.1 N hydrochloric acid (10 ml) and washed with diethyl ether (10 ml) twice. The aqueous layer was adjusted to pH 9.5 and extracted with ethyl acetate (12 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give a crude powdery product (149 mg). This powdery product was purified by reverse- phase preparative HPLC [carrier, ODS, YMC-pack, D-ODS-5; mobile phase 23% acetonitrile/0.01 M phosphate buffer, pH 6.3]; the fractions eluted in amounts of 720 to 830 ml were collected and concentrated to about 20 ml. The concentrated solution was adjusted to pH 10 and extracted with ethyl acetate (10 ml) twice. The obtained organic layer was washed with water and saturated saline (each 10 ml), dried over anhydrous sodium sulfate and concentrated to dryness to give compound 27 as powdery product (52 mg, a mixture of two isomers).

Elemental analys is ( for C19H24N2O2 O . 5H2O )

Calculated: C, 71.00; H, 7.84; N, 8.71

Found : C, 70.91; H, 7.47; N, 8.70 ¹ H NMR (δ ppm, in deuterochloroform, 300 MHz): 1.23/1.24 (3H, d, J=6.2 Hz), 1.34/1.34 (3H, s), 2.27 (IH, br), 2.48 (IH, d, J=11.4 Hz), 2.52 (3H, s), 2.61 (IH, ddd, J=14.6, 11.3, 1.6 Hz), 2.78 (IH, dd, J=11.4, 1.3 Hz), 3.00 (IH, br), 3.01 (IH, ddd, J=11.3, 5.7, 1.9 Hz), 3.46 (IH, dd, J=14.6, 5.7 Hz), 3.95/3.97 (IH, dd, J=14.3, 7.7 Hz), 4.07/4.08 (IH, dd, J=14.3, 3.5 Hz), 4.15 (IH, m) , 6.36/6.37 (IH, brs), 6.82 (IH, brs), 7.13-7.19 (3H, m) .

Example 28

Production of (5R, 8S)-l-carboxymethyl-9, 10-didehydro-6.8- dimethyl-8-ergolinol (Compound 19)

The compound 25 (118mg) as obtaind in Example 25 was dissolved in 1.0 N hydrochloric acid (120ml); this solution was stirred at room temperature for 48 hours. The solution was adjusted to PH 8.5 and washed with ethyl aretate (20ml) twice. Then the solution was concentrated to about 15ml. The concentrated solution was desalted with a desalting a- paratus (Microacylizer Gl, produced by Asahi Kasei, Japan), subjected to column chromatography with Diaion HP-20 (50- 100 mesh, 10ml), washed with water (50ml); the fractions eluted with 20-40% aqueous methanol were combined and concentrated to dryness to give a crude powdery product (45mg). To this powdery product, ethyl acetate and methanol were added; the precipitate was washed with ethyl acetate to give compound 19 as powdery product (19mg).

Example 29

Production of (5R, 8S)-didehydro-8-methyl-8-ergolinol

[Compound 29 (N-demethylsetoclavine) ]

N-Demethylagroclavine (5.08g) as obtained with the same method as described in Reference Example 4 was dissolved in 50% aquecous acetone (200ml) containing 2 N

sulfuric acid (22.6ml), followed by stirring at 70°C. To this solution, an aqueous solution (200ml) of potassium dichromate (6.70g) heated at 70°C was added; 1 minute later, 2N sulfuric acid (13.6ml) was added, followed by stirring at 70°C for 15 minutes. The reaction mixture was cooled to 0°C and adjusted to pH 2.5, after which it was stirred at room temperature for 1 hour and filtered through filter paper. The filtrate was adjusted to pH 6.0, mixed woth sodium hydrogen carbonate (12g), and 2 times extracted with chloroform-2-propanol mixture (3:1, 400ml). The obtained organic layer was dried over anhydrous sodium sulfate and concentrated to dryness to give as crude powdery product (4.4g). To this powdery product, ethyl acetate and diethyl ether were added; the obtained precipitate was washed with ethyl acetate-diethyl ether mixture to give powdery product (2.30g). The ethyl acetate-diethyl ether washings were concentrated to dryness to give a crude powder of compound 29 (1.46g, purity:50%). The obtained powder of compound 29 (4.4g) was subjected to silica gel column chramatography (50g, E.Merck, Art. 7734, Germany); the fractions eluted with methanol-acetone-hexane (5:30:7) containing 0.5% diethylamine were collected and concentrated to dryness. To this residue, acetone-ethyl acetate mixture was added; the obtained precipitate was washed with ethyl acetate to give compound 29 as powdery product (399 mg) . Elemental analysis (for C ₁₅ H ₁₆ N ₂ OO.4H ₂ O)

Calculated: C, 72.79; H, 6.84; N, 11.32 Found : C, 72.65; H, 6.67; N, 11.92 ^X H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.38 (3H, s), 2.20 (IH, br), 2.78 (IH, ddd, J=14.7, 11.6, 1.7 Hz), 2.91 (IH, d, J=12.1 Hz), 3.08 (IH, dd, J=12.1, 1.4 Hz), 3.22 (IH, dd, J=14.7, 6.0 Hz), 3.78 (IH, ddd, J=11.6, 6.0, 2.0 Hz), 6.41 (IH, brs), 6.91 (IH, t, J=1.7 Hz), 7.15-7.25 (3H, m), 7.98 (IH, brs).

Example 30

Production of (5R,8S)-9, 10-didehydro-6-ethyl-8-τnethyl-8- ergolinol (Compound 30).

A) The N-demethyl-N-ethylagroclavine (152 mg) as obtained in Reference Example 5 was dissolved in 50% aqueous acetone (6.0 ml) containing 2 N sulfric acid (0.30 ml), followed by stirring at 70°C. To this solution, an aqueous solution (177 ml) of potassium dichromate (177 mg) heated at 70°C was added; 1 minute later, 2 N sulfuric acid (0.39 ml) was added, followed by stirring at 70°C for 15 minutes. The reaction mixture was cooled to 0°C and adjusted to pH 2.5, after which it was stirred at room temperature for 1 hour and filtered through filter paper. The filtrate was adjusted to pH 9.0 and 2 times extracted with chloroform-2-propanol mixture (3:1, 20 ml). The obtained organic layer was dried over anhydrous sodium sulfate and concentrated to dryness to give as crude powdery product. To this crude powdery product, diethyl ether was added. The obtained mixture was filtered through filter paper. The filtrate was concentrated to dryness.

To this residue, diethyl ether was added; the obtained residue was washed with diisopropyl ether to give compound 30 as powdery product (40 mg). Elemental analysis (for Ci ₇ H ₂ o ₂ θ*0.8H ₂ θ)

Calculated: C, 72.21; H, 7.70; N, 9.91

Found : C, 72.15; H, 7.12; N, 9.91 ^X H NMR [ δ ppm, in deuterochloroform, 300 MHz): 1.12 (3H, t, J=7.2 Hz), 1.39 (3H, s), 2.58 (IH, d, J=11.2 Hz), 2.68 (IH, ddd, J=14.4, 11.1, 1.7 Hz), 2.86 (IH, dq, J=13.7, 7.1 Hz), 2.88 (IH, dd, J=11.2, 1.4 Hz), 3.05 (IH, dq, J=13.7, 7.3 Hz), 3.20 (IH, br) , 3.42 (IH, ddd, J=ll.l, 5.6, 1.9 Hz), 3.54 (IH, dd, J=14.4, 5.6 Hz), 6.41 (IH, brt, J=1.7 Hz), 6.91 (IH, t, J=1.8 Hz), 7.16-7.24 (3H, m) , 8.04 (IH, brs) .

B) A crude powder of compound 29 (424 mg, purity: 60%) as obtained with the same method as described in Example 29

was dissolved in acetonitrile (8.5 ml). To this solution, triethylamine (0.73ml), ethyl iodide (0.35 ml) was added, followed by stirring at 30°C for 3 hours. The reaction mixture was concentrated to dryness. To this residue, ethyl acetate (30 ml) was added. The obtained solution was washed with water and saturated saline (each 20 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery droduct (440 mg). The obtained crude powdery product was subjected to silica gel column chromatography (10 g, E.Merck, Art. 7734, Germany); the fractions eluted with acetone-hexane (20:80) containing 0.5% diethylamine were collected and concentrated to dryness. To this residue, diethyl ether-ethyl acetate was added. The precipitate was washed with diethyl ether to give compound 30 as powdery product (198 mg).

Example 31

Production of (5R,8S)-9,10-didehydro-8-methyl-6-propyl-8- ergolinol (Compound 31)

A crude powder of compound 29 (306 mg, purity: 50%) as obtained in Example 29 was dissolved in acetonitrile (7.6 ml) and stirred at 50°C. To this solution, triethylamine (0.44 ml) and propyl iodide (0.25 ml) were added; 1.5 hours later, triethylamine (0.44 ml) and propyl iodide (0.25 ml) were added; 3 hours later, triethylamine (0.44 ml) and propyl iodide (0.25 ml) were added, respectively, followed by strring for 1.5 hours. The reaction mixture was concentrated to dryness. To this residue, ethyl acetate (30 ml) was added. The obtained solution was washed with water and saturated saline (each 20 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (290 mg). The obtained crude powdery product was subjected to silica gel column chromatography (10 g, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-hexane (20:80) containing 0.5% diethylamine were collected and concentrated to

dryness. To this residue, diethyl ether-ethyl acetate- diisopropyl ether was added. The precipitate was washed with diisopropyl ether to give compound 31 as powdery product (150 mg) . Elemental analysis (for C ₁₈ H ₂₂ N ₂ O)

Calculated: C, 76.56; H, 7.85; N, 9.92

Found : C, 76.15; H, 7.87; N, 9.72 ^X H NMR (δ ppm, in deuterochloroform, 300 MHz): 0.97 (3H, t, J=7.4 Hz), 1.36 (3H, s), 1.58 (2H, m) , 2.54 (IH, d, J=11.2 Hz), 2.62 (IH, m) , 2.66 (IH, ddd, J=14.6, 11.2, 1.4 Hz), 2.89 (IH, m), 2.89, (IH, dd, J=11.2, 1.4 Hz), 3.05 (IH, br), 3.36 (IH, ddd, J=11.2, 5.5, 1.7 Hz), 3.54 (IH, dd, J=14.6, 5.5 Hz), 6.39 (IH, brs), 6.91 (IH, t, J=1.7 Hz), 7.14-7.23 (3H, m) , 7.91 (IH, brs).

Example 32

Production of (5R, 8S)-6-cyano-9,10-didehydro-8-methyl-8- ergolinol (Compound 32)

Compound A (159 mg) as obtained in Reference Example 2 was dissolved in dichloromethane (5.3 ml) and DMF (0.5 ml). To this solution, cynogen bromide (188 mg) was added, followed by stirring at 25°C for 22 hours. The reaction mixture was diluted with hexane (6 ml) and ethyl acetate (10 ml). The obtained solution was washed with saturated aqueous solution of sodium hydrogen carbonate and saturated saline (each 10 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (130 mg). The obtained crude powdery product was subjected to silica gel thin layer chromatography (Silica gel 60 F ₂₅₄ plate, 200x200x2mm, produced by E. Merck, Art. 5717, Germany), developing with methanol-acetone-chloroform (1:1:10). Then silica gel (RF value: 0.48-0.59) was pealed and eluted with methanol-acetone-chloroform (2:1:10). The obtained eluate was concentrated to dryness. To the residue, diethy lether and dichloromethane were added; the

precipitate was washed with diethyl ether to give compound 32 as powdery product (24 mg). Elemental analysis (for C ₁₆ H ₁₅ N ₃ OO.2H ₂ O)

Calculated: C, 71.46; H, 5.77; N, 15.63 Found : C, 71.46; H, 5.82; N, 15.43 ^λ E NMR (δ ppm, in deuterochloroform-methanol-d ₄ (9:l) , 300 MHz): 1.44 (3H, s), 3.08 (IH, ddd, J=14.3, 11.8, 1.7 Hz), 3.34 (IH, d, J=12.6 Hz), 3.50 (IH, dd, J=12.6, 1.0 Hz), 3.61 (IH, dd, J=14.3, 5.9 Hz), 4.16 (IH, ddd, J=11.8, 5.9, 2.0 Hz), 6.38 (IH, brs), 7.00 (IH, t, J=1.4 Hz), 7.18 (2H, m), 7.28 (IH, dd, J=6.5, 2.3 Hz).

Example 33

Production of (5R, 8S)-6-butyl-9,10-didehydro-8-methyl-8- ergolinol (Compound 33)

A crude powder of compound 29 (283 mg, purity: 44%) as obtained with the same method as described in Example 29 was dissolved in acetonitrile (7.5 ml). To this solution, triethylamine (0.39 ml) and butyl iodide (0.26 ml) were added, followed by stirring at 50°C for 2.5 hours. The reaction mixture was concentrated to dryness. To this residue, ethyl acetate (30 ml) was added. The obtained solution was washed with water and saturated saline (each 20 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (230 mg). The obtained crude powdery product was subjected to silica gel (10 g, E. Merck, Art. 7734, Germany) column chromatography; the fractions eluted with acetone-hexane (15:85) containing 0.5% diethylamine were collected and concentrated to dryness. To this residue, diethyl ether- diisopropyl ether-hexane mixture was added. The precipitate was washed with diisopropyl ether-hexan to give compound 33 as powdery product (52 mg). Elemental analysis (for C ₁₉ H ₂₄ N ₂ O)

Calculated: C, 76.99; H, 8.16; N, 9.45

Found : C, 76.55; H, 8.15; N, 9.33

^X H NMR ( δ ppm, in deuterochloroform, 300 MHz): 0.97 (3H, t, J=7.2 Hz), 1.37 (3H, s), 1.39 (2H, m) , 1.55 (2H, m) , 2.53 (IH, d, J=11.2 Hz), 2.62 (IH, ddd, J=13.2, 7.6, 5.6 Hz), 2.66 (IH, ddd, J=14.6, 11.3, 1.6 Hz), 2.89 (IH, dd, J=11.2, 1.1 Hz), 2.96 (IH, ddd, J=13.2, 8.6, 7.2 Hz), 3.06 (IH, br), 3.36 (IH, ddd, J=11.3, 5.5, 1.8 Hz), 3.54 (IH, dd, J=14.6, 5.5 Hz), 6.39 (IH, brs), 6.91 (IH, t, J=1.7 Hz), 7.16 (2H, m), 7.21 (IH, m) , 7.91 (IH, brs).

Example 34

Production of (5R, 8S)-6-allyl-9,10-didehydro-8-methyl-8- ergolinol (Compound 34)

The crude powder of compound 29 (243 mg, purity: 44%) as obtained in Example 29 was dissolved in acetonitrile (5.0 ml). To this solution, triethylamine (0.29 ml) and allyl bromide (0.15 ml) was added, followed by stirring at 25°C for 12 hours. The reaction mixture was concentrated to dryness. To this residue, ethyl acetate (20 ml) and 2% (w/v) aqueous solution of sodium hydrogen carbonate (15 ml) were added and mixed. The obtained organic layer was wased with water and saturated saline (each 20 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (240 mg). This crude powdery product was subjected to silica gel (10 g, E. Merck, Art. 7734, Germany) column chromatography; the fractions eluted with acetone-hexane (15:85) containing 0.5% diethylamine were collected and concentrated to dryness. To this residue diethyl ether was added. The precipitate was washed with diisopropyl ether to give compound 34 as powdery product (74 mg). Elemental analysis (for C ₁₈ H ₂₀ N ₂ O)

Calculated: C, 77.11; H, 7.19; N, 9.99

Found : C, 76.90; H, 7.18; N, 9.79 ¹ H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.36 (3H, s), 2.54 (IH, d, J=11.3 Hz), 2.69 (IH, ddd, J=14.5, 11.2 1.8 Hz), 2.94 (IH, dd, J=11.3, 1.4 Hz), 2.98 (IH, br), 3.28

(IH, dd, J=14.5, 8.1 Hz), 3.40 (IH, ddd, J=11.2, 5.6, 1.9 Hz), 3.57 (IH, dd, J=14.5, 5.6 Hz), 3.68 (IH, ddd, J=14.5, 5.0, 1.6 Hz), 5.22 (IH, brd, J=10.1 Hz), 5.28 (IH, dq, J=17.2, 1.1 Hz), 5.96 (IH, dddd, J=17.1, 10.1, 8.1, 5.0 Hz), 6.40 (IH, brs), 6.91 (IH, t, J=1.8 Hz), 7.16 (2H, m) , 7.21 (IH, m), 7.90 (IH, brs).

Example 35

Production of (5R,8S)-9,10-didehydro-6-isopropyl-8-methyl-

8-ergolinol (Compound 35)

A crude powder of compound 29 (235 mg, purity: 50%) as obtained with the same method as described in Example 29 was dissolved in acetonitrile (4.7 ml). To this solution, triethylamine (0.34 ml) and isopropyl iodide were added, followed by stirring at 25°C for 12 hours; after which potassium carbonate (135 mg), DMF (10 ml) and isopropyl iodide were added followed by stirring at 70°C for 10 hours, and further, at 60°C for 12 hours. The reaction mixture was concentrated to dryness. To this residue, ethyl acetate (20 ml) and 2% (w/v) aqueous solution of sodium hydrogen carbonate (15 ml) were added and mixed. The obtained organic layer was washed with water and saturated saline (each 20 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (232 mg). This crude powdery product was subjected to column chromatography with silica gel (10 g, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-hexane (15:85) containing 0.5% diethylamine were collected and concentrated to dryness. To this residue, diethyl ether was added. The precipitate was washed with diisopropyl ether to give compound 35 as powdery product (94 mg) . Elemental analysis (for C ₁₈ H ₂₂ N ₂ O)

Calculated: C, 76.56; H, 7.85; N, 9.92

Found : C, 76.06; H, 7.98; N, 9.70

^λ E NMR ( δ ppm, in deuterochloroform, 300 MHz): 0.95 (3H, d, J=6.5 Hz), 1.23 (3H, d, J=6.7 Hz), 1.37 (3H, s), 2.38 (IH, d, J=11.2 Hz), 2.62 (IH, ddd, J=15.7, 12.6, 1.7 Hz), 2.84 (IH, dd, J=11.2, 1.2 Hz), 3.15 (IH, br), 3.57 (2H, m) , 3.61 (IH, quint, J=6.6 Hz), 6.40 (IH, brs), 6.91 (IH, t, J=1.7 Hz), 7.16 (2H, m) , 7.21 (IH, m) , 7.92 (IH, brs).

Example 36

Production of (5R,8S)-6-cyclopropylmethyl-9,10-didehydro-8- methyl-8-ergolinol (Compound 36)

10

A crude powder of compound 29 (224 mg, purity: 50%) as obtained with the same method as described in Example 29 was dissolved in acetonitrile (3.5 ml) and DMF (1.0 ml). To this solution, triethylamine (0,34 ml), potassium - _c carbonate (129 mg) and cyclopropylmethyl bromide (0.19 ml) was added, followed by stirring at 60°C for 12 hours. The reaction mixture was concentrated to dryness. To the obtained residue, ethyl acetate (20 ml) and 2% (w/v) aqueous solution of sodium hydrogen carbonate (15 ml) were

20 added and mixed. The obtained organic layer was washed with water and saturated saline (each 20 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (248 mg). The residue was subjected to column chromatography with silica gel (10 g, E. Merck, Art. 7734, Germany); the fractions eluted with

25 acetone-hexane (15:85) containing 0.5% diethylamine were collected and concentrated to dryness. To this residue, diisopropyl ether was added. The precipitate was washed with diisopropyl ether to give compound 36 as powdery product (123 mg) .

30 Elemental analysis (for C ₁₉ H ₂₂ N ₂ O)

Calculated: C, 77.52; H, 7.53; N, 9.52 Found : C, 77.22; H, 7.44; N, 9.41 ^λ E NMR ( δ ppm, in deuterochloroform, 300 MHz): 0.19 (2H,

35 m), 0.53 (IH, dd, J=9.7, 8.2 Hz), 0.57 (IH, dd, J=9.7, 8.2 Hz), 0.98 (IH, m), 1.37 (3H, s), 2.66 (IH, ddd, J=13.6,

10.2, 1.6 Hz), 2.67 (IH, d, J=11.3 Hz), 2.69 (IH, dd, J=13.7, 6.3 Hz), 2.83 (IH, dd, J=13.7, 6.4 Hz), 3.09 (IH, dd, J=11.3, 1.4 Hz), 3.12 (IH, br), 3.50 (IH, ddd, J=10.2, 5.6, 1.8 Hz), 3.56 (IH, dd, J=13.6, 5.6 Hz), 6.41 (IH, brs), 6.91 (IH, t, J=1.9 Hz), 7.16 (2H, m) , 7.21 (IH, m) , 7.91 (IH, brs).

Example 37

Production of (5R, 8S)-9, 10-didehydro-6-(2, 2- diethoxyethyl)-8-methyl-8-ergolinol (compound 37)

A crude powder of compound 29 (214 mg, purity: 50%) as obtained in Example 29 was dissolved in acetonitrile (3.3 ml) and DMF (1.0 ml). To this solution, triethylamine (0.31 ml), potassium carbonate (124 mg) and 2, 2- diethoxyethyl bromide (0.28 ml) was added, followed by stirring at 60°C for 43 hours, and further, at 80°C for 22 hours. The reaction mixture was concentrated to dryness. To the obtained residue, ethyl acetate (20 ml) was added and mixed. The obtained organic solutiopn was washed with water and saturated saline (each 15 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (230 mg). The residue was subjected to column chromatography with silica gel (10 g, Kiselgel 60, 70-230 mesh, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-hexane (15:85) containing 0.5% diethylamine were collected and concentrated to dryness. To this residue, diisopropyl ether-hexane was added. The precipitate was washed with hexane to give compound 37 as powdery product (72 mg). Elemental analysis (for C ₂₁ H _{28 2} O ₃ )

Calculated: C, 70.76; H, 7.92; N, 7.86

Found : C, 70.57; H, 7.69; N, 7.72 ¹ H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.23 (3H, t, J=7.1 Hz), 1.25 (3H, t, J=7.0 Hz), 1.36 (3H, s), 2.68 (IH, ddd, J=14.2, 11.2, 1.6 Hz), 2.74 (IH, d, J=11.3 Hz), 2.83 (IH, dd, J=14.2, 5.1 Hz), 2.99 (IH, dd, J=11.3, 1.2

Hz), 3.05 (IH, br), 3.11 (IH, dd, J=14.2, 5.5 Hz), 3.47 (IH, ddd, J=11.2, 5.4, 1.7 Hz), 3.59 (IH, dd, J=14.2, 5.4 Hz), 3.59 (IH, dq, J=9.3, 7.1 Hz), 3.63 (IH, dq, J=9.3, 7.0 hz), 3.74 (IH, dq, J=9.3, 7.0 Hz), 3.76 (IH, dq, J=9.3, 7.1 Hz), 4.71 (IH, t, J=5.2 Hz), 6.38 (IH, brs), 6.91 (IH, t, J=1.8 Hz), 7.16 (2H, m) , 7.21 (IH, m) , 7.92 (IH, brs).

Example 38

Production of (5R, 8S)-9, 10-didehydro-8-methyl-6- tetrahydrofurfuryl-8-ergolinol (compound 38)

A crude powder of compound 29 (220 mg, purity: 50%) as obtained in Example 29 was diossolved in acetonitrile (3.4 ml) and DMF (1.0 ml). To this solution, triethylamine (0.32 ml), potassium carbonate (127 mg) and tetrahydrofurfuryl bromide (0.21 ml) was added, followed by stirring at 60°C for 94 hours. The reaction mixture was concentrated to dryness. To the obtained residue, ethyl acetate (20 ml) was added and mixed. The obtained organic solution was washed with water and saturated saline (each 15 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (248 mg). The residue was subjected to column chromatography with silica gel (10 g, Kiselgel 60, 70-230 mesh, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-hexane (20:80) containing 0.5 % diethylamine were collected and concentrated to dryness. To this residue, diisopropyl ether-hexane was added. The precipitate was washed with hexane to give compound 38 as powdery product (70 mg, a mixture of two isomers). Elemental analysis (for C ₂₀ ^H ₂ .N ₂ θ ₂ )

Calculated: C, 74.05; H, 7.46; N, 8.63

Found : C, 73.74; H, 7.38; N, 8.38 *H NMR ( δ ppm, in deuterochloroform, 300 MHz): 1.36/1.36 (3H,s), 1.65 (IH, m), 1.91 (2H, m) , 2.06 (IH, m) , 2.63/2.73 (IH, d, J=11.4 Hz), 2.66/2.70(lH, m) , 2.74 (1/2H, dd, J=14.1, 4.3 Hz), 2.77 (1/2H, dd, J=13.3, 3.9 Hz), 2.99

(1/2H, dd, J=13.3, 6.1 Hz), 2.99/3.03 (IH, dd, J=11.4, 1.2 Hz), 3.00 (IH, br), 3.05 (1/2H, dd, J=14.1, 7.1 Hz), 3.44 (IH, m), 3.50 (1/2H, dd, J=14.1, 5.6 Hz), 3.58 (1/2H, dd, J=14.3, 5.5 Hz), 3.77 (IH, m) , 3.91 (IH, m) , 4.13 (IH, m) , 6.38 (IH, brs), 6.90/6.90 (IH, t, J=1.8 Hz), 7.16 (2H, m) , 7.20 (IH, m), 7.92 (IH, brs).

Example 39

Production of (5R, 8S)-9, 10-didehydro-6-isobutyl-8-methyl-

8-ergolinol (compound 39)

A powder of compound 29 (107 mg) as obtained in Example 29 was dissolved in acetonitrile (2.5 ml) and DMF (1.0 ml). To this solution, potassium carbonate (247 mg) and isobutyl bromide (0.10 ml) were added, followed by stirring at 80 ^β C for 48 hours; after which isobutyl bromide 0.05 ml was added followed by stirring at 70°C for 24 hours. The reaction mixture was concentrated to dryness. To the obtained residue, ethyl acetate (20 ml) was added and mixed. The obtained organic solution was washed with water and saturated saline (each 15 ml), dried over anhydrous sodium sulfate, and concentrated to dryness to give a crude powdery product (150 mg). The residue was subjected to column chromatography with silica gel ( 10 g, Kiselgel 60, 70-230 mesh, E. Merck, Art. 7734, Germany); the fractions eluted with acetone-hexane (15:85) containing 0.5 % diethylamine were collected and concentrated to dryness. To this residue, diisopropyl ether-hexane was added. The precipitate was washed with hexane to give compound 39 as powdery product (50 mg). Elemental analysis (for CιgH _{2 2} θ-0.3H ₂ θ)

Calculated: C, 76.12; H, 8.27; N, 9.34

Found : C, 76.18; H, 8.19; N, 9.20 E NMR ( δ ppm, in deuterochloroform, 300 MHz): 0.96 (3H, d, J=6.6 Hz), 1.03 (3H, d, J=6.5 Hz), 1.36 (3H, s), 1.92 (IH, m), 2.24 (IH, dd, J=12.8, 4.6 Hz), 2.45 (IH, d, J=11.2 Hz), 2.65 (IH, ddd, J=14.7, 11.3, 1.8 Hz), 2.67 (IH, dd,

J=12.8, 9.7 Hz), 2.91 (IH, dd, J=11.2, 1.2 Hz), 2.97 (IH, brs), 3.26 (IH, ddd, J=11.3, 5.4, 1.7 Hz), 3.52 (IH, dd, J=14.7, 5.4 Hz), 6.39 (IH, brs), 6.90 (IH, t, J=1.7 Hz), 7.16 (2H, m), 7.21 (IH, m) , 7.89 (IH, brs).

The compounds obtained in Examples 4 through 39 above are shown in Table 1.

Table 1

Compound Example Number Number 2 Y2 z

4* 4 H Methyl H

5* 5 H Methyl Methyl

6* 6 H Methyl Ethyl

7* 7 H Methyl n-Propyl

8A* 8 H Methyl Allyl

9* 9 H Methyl Benzyl

10* 10 Methyl Methyl H

11* 11 Methyl Methyl Methyl

12* 12 Ethyl Methyl H

13* 13 Allyl Methyl H

14* 14 n-Hexyl Methyl H

15* 15 Isopropyl Methyl H

16 16 Benzyl Methyl H

17 17 Ethyl Methyl H

18 18 CH ₂ CONH ₂ Methyl H

19 19 CH ₂ C0 ₂ H Methyl H

20 20 n-Propyl Methyl H

21 21 CH ₂ OCH ₃ Methyl H

22A 22 CH ₂ OCH ₂ CH ₂ OCH ₃ Methyl H

23 23 n-Butyl Methyl H

24 24 CF=CHI Methyl H

25 25 CH ₂ C0 ₂ Bu ^t Methyl H

26 26 n-Pentyl Methyl H

27 27 2-Hydroxypropyl Methyl H

29 29 H H H

30 30 H Ethyl H

31 31 H n-Propyl H

32 32 H CN H

33 33 H n-Butyl H

34 34 H Allyl H

35 35 H Isopropyl H

36 36 H Cyclopropyl¬ H methyl

37 37 H 2, 2- H Diethoxyethyl

38 38 H Tetrahydro¬ H furfuryl

39 39 H Isobutyl H

Racemate

Using the mouse substance P spinal intrathecal injection method (SP-i.t. method), mouse formalin method, rat hot plate method and mouse acetic acid writhing method, the analgic effects of the compounds produced in Examples and Reference Examples were determined.

For the mouse substance P spinal intrathecal injection method (SP-i.t. method), mouse formalin method and mouse acetic acid writhing method, analgic effects are expressed as 50% suppressive doses (ID ₅₀ ). ID ₅₀ values and 95% confidence limits were obtained from the linear regression

line of the log dose-response curves. Analgic effect increases as the ID ₅ 0 value decreases.

For the rat hot plate method, analgic effects are expressed as minimum effective doses (MED) . MED significant differences were tested by Dunnett's test. Analgic effect increases as the MED value decreases.

Experimental Example 1

Analgic activity in the mouse substance P spinal intrathecal injection method (SP-i.t. method)

Method

Male Slc/ICR mice (5 weeks old, 6 - 10 animals per group) were used. Test drugs were injected into the spinal subarachnoid space in accordance with the method described by Hyden, ilcox et al. in European Journal of Pharmacology, Vol. 67, p. 313 (1980). Five μl of a physiological saline solution of substance P containing 0.25% of 2 N hydrochloric acid (2.0 mg/liter) was injected into the spinal subarachnoid space . Substance P elicited caudally directed biting and licking. The numbers of reciprocal biting and licking were counted for 1 minute after substance P injection; this value was used as an index of pain reaction. Compound 3A or 4, in a mixed solution with substance P, was co-injected into the spinal subarachnoid space [T. Doi et al., European Journal of Pharmacology, Vol. 137, p. 227 (1987)]. Results

Table 2 shows the ID ₅₀ values ( g/mouse) of compounds 3A and 4, as determined by the mouse substance P spinal intrathecal injection method (SP-i.t. method).

Table 2

ID ₅₀ ( g/mouse)

3A 0.14

4 0.27

Experimental Example 2

Analgic activity in the mouse formalin method

Method

Male Slc/ICR mice (5 weeks old, 10 animals per group) were used. Compounds 3A, 4, 17, A and morphine (control) were orally administered. Thirty minutes later, 10 μl of a 0.5% aqueous solution of formalin was subcutaneously administered to a hind leg paw. Cumulative time of behavior (biting or licking at injected sites) was measured for 5 minutes after subcutaneous administration; this value was used as an index of pain reaction. Results

Table 3 shows the ID ₅ 0 values (mg/kg) of compounds 3A, 4, 17 and A and morphine, as determined by the mouse formalin method.

Table 3

ID ₅₀ (mg/kg)

3A 8.4

4 4.6

17 3.5

A 1.8

Morphine 40.2

Experimental Example 3 Analgic activity in the rat hot plate method

Method

Male Jcl:Wistar rats (5 weeks old, 6 animals per group) were used. Compounds 4 and A and morphine (control) were subcutaneously or orally administered. Thirty minutes after the administration, each rat was placed on a 50°C copper plate in a glass cylinder of 15 cm diameter and 20 cm height; latent time of behavior (hind leg licking, or jumping up) was measured, with a cutoff time of 60 seconds. Results

Table 4 shows the MED values (mg/kg) of compounds 4 and A and morphine, as determined by the rat hot plate method.

Table 4

ID ₅₀ (mg/kg)

Subcutaneous Oral

4 0.625 20

A 0.313 5

Morphine 2.5 50

Experimental Example 4

Analgic activity in the mouse acetic acid writhing method

Method

Male Slc/ICR mice (4 weeks old, 10 animals per group) were used. Compounds 4, 17 and A were orally administered, Thirty minutes later, a 0.6% aqueous solution of acetic acid was injected intraperitoneally at 0.1 ml/10 g body weight. Writhing in response to acetic acid stimulation was counted for 20 minutes after the intraperitoneal administration; this value was used as an index of pain reaction [E. Siegmund et al., Proceedings of the Society for Experimental Biology and Medicine, Vol. 95, p. 729].

Results

Table 5 shows the ID ₅ 0 values (mg/kg) of compounds 4, 17 and A, as determined by the mouse acetic acid writhing method.

Table 5

ID ₅₀ (mg/kg )

4 1 . 19

17 2 .82

A 0 .91

Experimental Example 5 Toxicity test

In an acute toxicity study in mice, compound 4 did not cause death at 200 mg/kg, whether intraperitoneal or oral administration.

The compound of the present invention has excellent analgic activity, with low toxicity, and is useful as an analgic agent for patients with rheumatoid arthritis, neuralgia, osteoporosis, terminal cancer etc.

Preparation Example 1

Compound 4 as obtained in Example 4 and the following components were mixed and packed in gelatin capsules to yield capsules each containing 30 mg of compound 4.

Compound 4 30 mg

Lactose 100 mg

Corn starch 40 mg

Magnesium stearate 10 mg

Total 180 mg

Preparation Example 2

Compound 4 as obtained in Example 4, lactose, corn starch (half the amount shown below) and hydroxypropyl cellulose were mixed; this mixture was kneaded and granulated with water. After vacuum drying, the granules were mixed with a mixture of magnesium stearate and corn starch (half the amount shown below). The resulting mixture was subjected to compressive shaping to yield tablets of the following composition.

Compound 4 60 mg

Lactose 68.4 mg

Corn starch 65 mg

Hydroxypropyl cellulose 6 mg

Magnesium stearate 0.6 mg

Total 200.0 mg

Preparation Example 3

Compound 4 as obtained in Example 4 was dissolved in physiological saline containing 30% (w/v) polyethylene glycol 400 to give a 0.05% solution of compound 4. After sterilization and filtration, this solution was dispensed to vials at 30 ml per vial, to yield an intravenous injection preparation containing 15 mg of compound 4 per vial.

Preparation Example 4

Compound 4 as obtained in Example 4 was dissolved in MIGLYOL 812 [caprylic acid/capric acid triglyceride, HULS AKTIENGESELLSCHAFT, Germany] to 10 mg/ml (1% w/v) to give a uniform solution. This solution was packed in nitrogen- replaced vials at 20 ml per vial, to yield a preparation containing compound 4.

Preparation Example 5

Compound A as obtained in Reference Example 2 and the following components were mixed and packed in gelatin

capsules to yield capsules each containing 30 mg of compound A.

Compound A 30 mg

Lactose 110 mg

Corn starch 30 mg

Magnesium stearate 10 mg

Total 180 mg

Preparation Example 6

Compound A as obtained in Reference Example 2, lactose, corn starch (half the amount shown below) and hydroxypropyl cellulose were mixed; this mixture was kneaded and granulated with water. After vacuum drying, the granules were mixed with a mixture of magnesium stearate and corn starch (half the amount shown below). The resulting mixture was subjected to compressive shaping to yield tablets of the following composition.

Compound A 60 mg

Lactose 68.4 mg

Corn starch 65 mg

Hydroxypropyl cellulose 6 mg

Magnesium stearate 0.6 mg

Total 200.0 mg

Preparation Example 7

Compound A as obtained in Reference Example 2 was dissolved in physiological saline containing 30% (w/v) polyethylene glycol 400 to give a 0.05% solution of compound A. After sterilization and filtration, this solution was dispensed to vials at 30 ml per vial, to yield an intravenous injection preparation containing 15 mg of compound A per vial.

Preparation Example 8

Compound A as obtained in Reference Example 2 was dissolved in MIGLYOL 812 [caprylic acid/capric acid triglyceride, HULS AKTIENGESELLSCHAFT, Germany] to 10 mg/ml (1% w/v) to give a uniform solution. This solution was packed in nitrogen-replaced vials at 20 ml per vial, to yield a preparation containing compound A.

Compound (I) or salt thereof of the present invention possesses excellent analgic activity, and can be safely used as an analgic agent for patients with rheumatoid arthritis, neuralgia, osteoporosis, terminal cancer etc.