JPH0739409B2 - Novel forskolin derivative - Google Patents

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel forskolin derivative having a positive inotropic effect, a hypotensive effect and an adenylate cyclase activating effect.

[0002]

2. Description of the Related Art Forskolin having positive inotropic action, hypotensive action and adenylate cyclase activating action has already been known [JP-A-52-79015, Drug Research, 31 1248 (198).
1)].

[0003]

However, forskolin is poorly water-soluble and dissolves only 0.0026% in water at room temperature at room temperature. Therefore, a water-soluble derivative is desired because it requires devising a formulation.

[0004]

Therefore, as a result of various investigations, the present inventors have found that the general formula (I)

[0005]

[Chemical 2]

(In the formula, when R ¹ is a hydrogen atom and R ⁴ is a vinyl group, an ethyl group or a cyclopropyl group,

(1) One of R ² and R ³ is a partial structural formula CO (CH ₂ ) mNR ⁵ R ⁶ (wherein R ⁵ and R ⁶ are hydrogen atoms, lower alkyl groups, or R ⁵ and R ⁶ Is a lower alkylene group which may be bonded to each other and may contain an oxygen atom or a nitrogen atom in the bonding chain, and m is an integer of 1 to 5), and the other is a hydrogen atom or a partial structural formula CO (C
H ₂ ) nX (where X is a hydrogen atom or a group represented by the formula NR ⁷ R ⁸ (wherein R ⁷ and R ⁸ are a hydrogen atom, a lower alkyl group or R ⁷ and R ⁸ are bonded to each other, Is a lower alkylene group which may contain an oxygen atom or a nitrogen atom in the bonding chain, and n is an integer of 1 to 5, provided that R ³ is an acetyl group and R ⁴ is a vinyl group. And R ² is a group other than a dimethylaminopropionyl group).

(2) R ² is a hydrogen atom or --COCH ₂
When CH ₂ CO ₂ H, R ³ is a hydrogen atom, and is of the formula —CO
_{CH 3, -COCH 2 CH 2 CH} 2 CO 2 H or -C,
OCH (OH) CH ₂ OH (provided that when R ² is a hydrogen atom, R ³ is a hydrogen atom or a group other than COCH ₃ ).
Shows

II. R ¹ is a partial structural formula CO (CH ₂ ) pC
A group represented by O ₂ H or CO (CH ₂ ) qNR ⁹ R ¹⁰ (wherein R ⁹ and R ¹⁰ are hydrogen atoms or a lower alkyl group, p is 0 to 5 and q is an integer of 1 to 5). R ² represents a hydrogen atom, R ³ represents an acetyl group, and R ⁴ represents a vinyl group], wherein the forskolin derivative and a physiologically acceptable salt thereof are acidic, neutral, or basic. It has been found that it is more water soluble than forskolin and has positive inotropic effect, hypotensive effect, and adenylate cyclase activating effect.

The present invention has been completed based on the above findings.

Examples of the lower alkyl group in the above general formula (I) include alkyl groups having 1 to 4 carbon atoms such as methyl group, ethyl group, propyl group and butyl group. Further, the lower alkylene group which may contain an oxygen atom or a nitrogen atom in the bonding chain is, for example,-(CH ₂ ) ₃ -,-(-
CH ₂ −) ₄ −, − (− CH ₂ −) ₅ −, − (− CH _{2
-) 2 -NH- (CH 2 -} ) 2 -, (CH 2) 2 -O-
Examples thereof include (CH ₂ −) ₂ − and C _{3 to} C ₅ .

As --CO-(-CH ₂ ) _n --NR ⁵ R ⁶ in R ² or R ³ of the above general formula (I), for example, dimethylaminoacetyl group, diethylaminoacetyl group, diethylaminopropionyl group, butylaminoacetyl group, Examples thereof include aminopropionyl group, aminobutyryl group, methylaminopropionyl group, dimethylaminopropionyl group, dimethylaminobutyryl group, pyrrolidinobutyryl group, pyrrolidinoacetyl group, piperazinoacetyl group and morpholinoacetyl group. Also, the other -C
O - (- CH ₂ -) as the _n -X such as acetyl group,
Examples thereof include a propionyl group, a butyl group, and the same aminoacyl groups as the above various aminoacyl groups. As a preferred compound, R ³ is a lower alkylcarbonyl group such as acetyl group, R ² is —CO (CH ₂ ) _n —NR ⁵ R ⁶ and n =
1 to 4, compounds in which R ⁵ and R ⁶ are lower alkyl groups can be mentioned.

Further, the present invention is preferably the above formula (I)
General formula (I ′) which limits the represented compounds

[Chemical 3] [In the formula, I. R ¹ is a hydrogen atom, R ⁴ is a vinyl group, an ethyl group or
When it is a cyclopropyl group , (1) R ² is a partial structural formula CO (CH ₂ ) mNR ⁵ R ⁶ (
Here, R ⁵ and R ⁶ are a hydrogen atom, a lower alkyl group, or R
⁵ and R ⁶ are bonded to each other, and an oxygen atom or a nitrogen atom is present in the bonding chain.
Is a lower alkylene group which may include
Is an integer of 5) and R ³ is a lower alkylcarbonyl
Group (provided that R ³ is an acetyl group and R ⁴ is a vinyl group)
In some cases, R ² is a group other than a dimethylaminopropionyl group.
Is. ) Or (2) R ² represents a hydrogen atom and R ³ represents —COCH (O
H) CH ₂ OH or (3) R ² represents —COCH ₂ CH ₂ CO ₂ H and R ³
Is _{-COCH 3, -COCH 2 CH 2 CH} 2 CO 2 H,
Or II showing a -COCH (OH) CH ₂ OH. R ¹ is
Group represented by partial structural formula CO (CH ₂ ) pCO ₂ H
(Where p is an integer of 0 to 5), R ²
Is a hydrogen atom, R ³ is an acetyl group, and R ⁴ is a vinyl group.
] And its physiology
Related to a permissible salt. Next, a method for producing the compound represented by formula (I) will be described. 1. In the general formula (I), a compound in which R ¹ and R ² are both hydrogen atoms and R ³ is —CO (CH ₂ ) _m NR ⁵ R ⁶ can be produced by any of the following methods.

(1) 7-deacetylforskolin (R
^{1 = R 2 = R 3 =} H, R 4 = -CH = CH 2), or 7-deacetyl -14,15- dihydro Fuorusu choline ^{(R 1 = R 2 = R} 3 = H, R 4 = CH 2 CH ₃ ) or 13-cyclopropyl-7-deacetyl-14,1
5 Dinorphouscholine (R ¹ = R ² = R ³ = H, R ⁴
= Cyclopropyl group), a compound represented by the general formula (II) R ⁵ R ⁶ N (CH ₂ ) _m CO ₂ H (II) [wherein R ⁵ , R ⁶ and _m are the same as above] or a reaction thereof. by condensing sex derivative, to give a compound of _{R 3 = -CO (CH 2)} m NR 5 R 6.

(2) 7-deacetylforskolin 7-
Deacetyl-14,15-dihydroforskolin or 13-cyclopropyl-7-deacetyl-14,15
General formula (III) Y-(-CH _2- ) _m -COOH (III) in which dinorphoruscholine is represented, wherein Y is a halogen atom or alkyl or aryl
Indicates Le sulfonyloxy group, _m is condensed a compound or a reactive derivative thereof represented by the same] as above, and then in the general formula ^{(IV) R 5 R 6 N} -H (IV) [wherein, R ^5, R ⁶ Is the same as the above], and R ³ ═—CO (C
H ₂ ) _m NR ⁵ R ⁶ compounds can be obtained.

(3) 7-deacetylforskolin or 7-deacetyl-14,15-dihydroforskolin or 13-cyclopropyl-7-deacetyl-1
4,15- Gino Le Fuorusu choline general formula _{(V) CH = CH 2 -} (- CH 2) m'-2 -COOH (V) wherein, _{m 'is} an integer of 2-4] represented by Compound or a reactive derivative thereof is condensed, and then the above general formula
The compound represented by (IV) is reacted.

In the condensation of above (1) to (3), the general formula
(II), (III), when using the carboxylic acid itself as the compound (V), in a solvent such as benzene, chloroform, ether, ethyl acetate, dicyclohexylcarbodiimide,
Using dicyclohexylcarbodiimide + 4-dimethylaminopyridine, carbonyldiimidazole, diphenylphosphoryl azide, or the like, for example, −20 ° C. to 200 ° C.
Temperature is usually 0.5 to 7 near the boiling point of the solvent under ice cooling.
It is preferable to carry out the condensation reaction for 2 hours, preferably 2 to 48 hours, which gives preferable results.

Examples of the compound represented by the general formula (II) include dimethylaminoacetic acid, n-butylaminoacetic acid, diethylaminoacetic acid, pyrrolidinoacetic acid, piperazinoacetic acid, monofolinoacetic acid, dimethylaminopropionic acid, and pyrrolidinobutyric acid.

Examples of the compound represented by the general formula (III) include halogenoacetic acid such as chloroacetic acid and bromoacetic acid, chloropropionic acid, chlorobutyric acid, chlorovaleric acid, methanesulfonyloxyacetic acid, p-toluenesulfonyloxybutyric acid and methanesulfonyloxykylate. Examples include herb acid.

Examples of the compound of the general formula (V) include acrylic acid, methacrylic acid, vinyl acetic acid, allyl acetic acid and the like.

In the condensation of above (1) to (3), the general formula
When the reactive derivative of (II), (III) or (V) is used, it can be obtained by reacting with a base such as pyridine or triethylamine in a solvent such as benzene, chloroform, ether or ethyl acetate. . It is preferable to carry out the reaction under ice-cooling for about 2 to 48 hours near the boiling point of the solvent.

Examples of the reactive derivative include acid halides, acid anhydrides, mixed acid anhydrides, and Leuhi anhydride.

In the production methods (2) and (3), the general formula
The reaction of (IV) with the amine is carried out by stirring in a solvent such as dichloromethane under ice cooling for about 0.5 to 5 hours.

Examples of the amine of the general formula (IV) include dimethylamine, butylamine, diethylamine, pyrrolidine, piperazine and morpholine.

(4) No. 26 by reacting 7-deacetylforskolin with glutaric anhydride in a solvent such as benzene, chloroform, ether or ethyl acetate using a base such as pyridine or triethylamine. The compound is obtained. The reaction is from 2 to 4 near the boiling point of the solvent under ice cooling.
8 hours gives desirable results.

(5) The compound of No. 27 is obtained by condensing 7-deacetylfluoroskolin with glyceric acid or its reactive derivative. When using glyceric acid itself for condensation, use benzene, chloroform,
Dicyclohexylcarbodiimide, dicyclohexylcarbodiimide + 4-in a solvent such as ether or ethyl acetate
Using dimethylaminopyridine, carbonyldiimidazole, diphenylphosphoryl azide, or the like, it is preferable to carry out the condensation reaction for 2 to 48 hours, for example, under ice cooling and around the boiling point of the solvent.

When a reactive derivative is used during condensation, it can be obtained by reacting with a base such as pyridine or triethylamine in a solvent such as benzene, chloroform, ether or ethyl acetate. It is preferable to carry out the reaction under ice-cooling for about 2 to 48 hours near the boiling point of the solvent. Examples of the reactive derivative include acid halides, acid anhydrides, mixed acid anhydrides, and Leuhi's anhydride.

2. In the general formula (I), R ² is a group other than a hydrogen atom, and R ¹ and R ³ are both hydrogen atoms, and the compound is a 7-substituted forskolin or 7-substituted-14 obtained by the above method 1. 15-dihydroforskolin or 7-substituted-13-cyclopropyl-14,1
It can be obtained by rearranging the substituent at the 7-position of 5-dinorforskolin to the 6-position.

The rearrangement reaction is carried out by using dimethyl sulfoxide, N,
In an organic solvent such as N-dimethylformamide, methanol, acetone, acetonitrile dioxane, or tetrahydrofuran, or in a mixed solvent of each solvent and water,
Preferably, in a mixed solvent of N, N-dimethylformamide or acetonitrile and water, 0.1 to 10 equivalents, preferably 1 to 3 equivalents of an alkali metal hydride, hydroxide or carbonate, for example, hydroxide. Inorganic bases such as sodium, potassium hydroxide, potassium carbonate, sodium hydride, tri-lower alkylamines such as triethylamine, and organic bases such as 1,8-diazebicyclo [5,4,0] -7-undecene at the 7-position It is carried out by acting on recombinant forskolin. The reaction is about -20 ° C to about 2
Temperature of 00 ° C Usually from under ice-cooling to around the boiling point of the solvent for 1 minute to
It is carried out for 48 hours, preferably 30 minutes to 1 hour at room temperature.

The intermediate obtained in the condensation step (2) or (3) of 1 above is treated in the same manner as in 2 above, and then the above formula
It can also be obtained by reacting with the amine of (IV).

3. In the general formula (I), R ² is —COC.
A compound which is H ₂ CH ₂ COOH and in which both R ¹ and R ³ are hydrogen atoms can be obtained as follows.
That is, in the method of 1 (4) above, succinic anhydride is used instead of glutaric anhydride to obtain 7-substituted forskolin, and then the substituent at the 7-position is rearranged to the 6-position.

The rearrangement reaction is dimethyl sulfoxide, N, N
-In a polar solvent such as dimethylformamide, methanol, acetone, acetonitrile or in a mixed solvent of each solvent and water, preferably in a mixed solvent of N, N-dimethylformamide or acetonitrile and water,
0.1 to 10 equivalents, preferably 1 to 3 equivalents of inorganic base such as sodium hydroxide, potassium hydroxide, potassium carbonate, sodium hydride, or triethylamine, 1,8
It is carried out by reacting an organic base such as -diazabicyclo [5,4,0] -7-undecene with the 7-position-substituted forskolin. The reaction is carried out under ice-cooling for about 1 minute to 48 hours near the boiling point of the solvent, preferably at room temperature for 30 minutes to 1 hour.

4. In the general formula (I), R ¹ is a hydrogen atom, and R ² and R ³ are both groups other than a hydrogen atom, and can be obtained by any of the following methods. (1) 6-Substitution-7- obtained by the above method 2
Deacetylforskolin, 6-substituted-7-deacetyl-14,15-dihydroforskolin or 6-substituted-13-cyclopropyl-7-deacetyl 14,15-
Gino Le Fuorusu choline or the similar obtainable by the process of 6-alkanoyl-7-deacetyl - Fuorusu choline general formula _{Y - (- CH 2 -)} m -COOH (VI) wherein, Y, _m is said The same as the above] or a reactive derivative thereof is condensed in the same manner as in the above 1 (1).

Examples of the compound of the general formula (VI) include acetic acid, propionic acid, butyric acid, and the compounds exemplified as the compound of the general formula (II).

(2) In the case of a compound in which the substituent at the 7-position is a nitrogen-containing group, 7- used in 1 (2) or (3) above
6-substituted-7-deacetylforskolin 6 obtained by the above method 2 instead of deacetylforskolin 6
-Substituted-7-deacetyl-14,15-dihydroforskolin or 6-substituted-13-cyclopropyl-1
It can be obtained by carrying out the same reaction as 1 (2) or (3) above using 4,15-dinorforskolin or 6 alkanoyl-7-deacetylforskolin obtained by the same method as in 2 above.

(3) In the general formula (I), R ¹ is a hydrogen atom, R ² is —COCH ₂ CH ₂ COOH,
And R ³ is COCH ₃ , COCH ₂ CH ₂ CH ₂ COO
H or —COCH (OH) CH ₂ OH,
As the first step, 6- (2-carboxyethylcarbonyl) -7-deacetylphoruscholine is synthesized by the method described in 3 above, and then acetic acid or a reactive derivative thereof is reacted, or A) or (5) to carry out acylation to obtain the desired product.

In the reactions 1 to 4, instead of the raw material such as 7-deacetylphoruscholine, the 1-position hydroxyl group is an acyl group such as an acetyl group, an ether group such as a methyl ether group, and t-. The compound of the general formula (I) of the present invention can also be obtained by using a compound protected with a silyl ether group such as butyldimethylsilyl ether group and removing the protecting group in the final step.

5. In the general formula (I), 6 and / or 7-di (or mono) -substituted-13-cyclopropyl or 1 in which R ⁴ is an ethyl group or a cyclopropyl group.
4,15-Dihydro-forskolin is obtained by hydrogenating the 6- and / or 7-di (or mono) -substituted-forskolin produced by the above-mentioned production methods 1 to 3 or by reacting diazomethane with palladium acetate or the like as a catalyst. It can be manufactured by

6. In the general formula (I), R ¹ is —CO
(CH ₂₎ pCOOH or -CO- (CH ₂₎ qNR
⁹ is R ¹⁰ is compounds in general formula (I) compound is Oite R ¹ is a hydrogen atom, the formula HOOC (CH ₂₎ pCOOH or _{^{R 9 R 10 N (CH 2}} ) a carboxylic acid or represented by qCOOH It is obtained by reacting a reactive derivative in a solvent at -20 ° C to 200 ° C.

In the reactions 1 to 6, the carboxyl groups and hydroxyl groups on R ¹ , R ² and R ³ are ester groups such as methyl ester group and ethyl ester group, acyl groups such as acetyl group, methyl ether group and acetonide. A compound protected with an ether group such as a group and a silyl ether group such as a t-butyldimethylsilyl ether group is used, and the amino group on R ¹ , R ² and R ³ is an acyl group such as an acetyl group, t The compound of the general formula (I) of the present invention can also be obtained by using a compound protected with a silyl ether group such as -butyldimethylsilyl ether group and removing the protecting group in the final step.

Unless otherwise specified, the above-mentioned reaction is preferably carried out in an organic solvent or a mixed solvent of them and water. The compound of the present invention is purified and isolated from the reaction solution by a usual method, and the reaction conditions and It is obtained as a free base, free acid or salt depending on the method of treatment.

If desired, the free base and free acid can be converted into salts by conventional methods. As the salt, hydrochloride,
Inorganic acid salts such as hydrobromide, sulfate, phosphate, etc. or formate, acetate, fumarate, maleate, citrate, tartrate, lactate, methanesulfonate, etc. It can be an organic acid salt. The free acid is a metal salt such as sodium salt, potassium acid, magnesium salt, calcium salt, or 4
It can be an organic basic acid such as a primary ammonium salt or a pyridinium salt.

Among the compounds of the present invention, R ² or R ³
Since those having an asymmetric carbon atom in theory have optical isomers, the present invention includes those optical isomers. The optical isomers can be separated by a known method such as chromatography or fractional crystallization.

The compounds of the present invention produced by the above method are shown in Table 1. In addition, in Table 1, the example compounds are
Compound No. 10-12, 15, 18, 20, 27, 2
9, 31-33, 38, 39, 41, 43-46.
The reference example compound is a compound No. other than the example compound. of
Show things.

[0045]

[Table 1]

[0046]

[Table 2]

[0047]

[Table 3]

[0048]

[Table 4]

[0049]

Next, the action of the compound of the present invention will be described. 1. Positive inotropic effect, hypotensive effect Under the anesthesia of pentobarbital sodium in both beagles and adult dogs of both beagles and males and females, polyethylene tube was inserted from the carotid artery into the left ventricle for measurement of left ventricular pressure, or Insert a polyethylene tube into the femoral artery for blood pressure measurement. These polyethylene tubes are connected to a pressure transducer and continuously recorded on a recorder via a pressure strain gauge. In addition, the rising speed of the left ventricular pressure (dp /
dt) is obtained by a differentiator and continuously recorded in the same manner, and this is used as an index of inotropic effect.

The effect of the drug is expressed as a relative value to the maximum response given to each index of forskolin. That is, dp / when intravenously administered 30 μg / kg of forskolin
The maximum change in dt rise and mean blood pressure (MBP) fall is 1,
300 μg / kg or 30 μg / kg of test compound, respectively
The maximum change when kg is intravenously administered is expressed as a relative value.

2. Adenylate cyclase activating action As an adenylate cyclase standard, a membrane fraction obtained from a homogenate of guinea pig myocardium was used [G. I. Drum
mond, D.M. L. Severson, L. Duncan, J. Biol. Chem., 2
46, 4166 (1971)].

The measurement is carried out by the method of Salomon et al. [Y. Salomon, C.
Londos, M. Rodbell, Anal. Bioc-hem., 58.541
(1974)], labeled cAMP produced by adenylate cyclase is measured using labeled ATP as a substrate. The reaction solution was 5 mM MgCl ₂ , 20 mM creatine phosphate, 100 U / ml creatine phosphokinase, 1
mM, cAMP, 1 mM [ ¹⁴ C (U)] ATP (about 7 c
pm / pmol) and forskolin or test compound (1 μM) in 25 mM Tris.HCl (pH 7.
In 5), the enzyme preparation is added so that the final volume becomes 100 μl, and the reaction is started. The amount of enzyme is 15 as a membrane protein
It is set to 0 to 200 μg / 100 μl. After reacting at 37 ° C. for 10 minutes, a reaction stop solution (2% sodium dodecyl sulfate (SDS), 40 mM ATP, 1.4 mM cAM
After stopping the reaction by adding 100 μl of P, pH 7.5), 50 μl of [ ³ H] cAMP (about 20000 cpm) is added in order to know the recovery rate of cAMP. After this, cAMP is separated using a Dowex 50 resin column and a neutral alumina column, and the radioactivity is measured.

The activating effect of the test compound (1 μM) on adenyl cyclase is expressed as a percentage of the activating effect of forskolin (1 μM). [N = 4 (No. 10, 1
7 is n = 6 No. 38, 39, 41 is n = 2), average value ±
Standard error]. Next, Table 2 shows the results of the above 1 and 2 for the representative compounds of the present invention.

[0054]

[Table 5]

[0055]

[Table 6]

[0056]

[Table 7]

3. Solubility As a result of measuring the solubility of the compounds of the present invention in water, all the compounds exhibited a solubility of 0.1% or more. Since the solubility of forskolin in water is 0.0026%,
The compound of the present invention has a solubility of 38 times or more that of forskolin.

[0058]

[Effect] As described above, the compound of the present invention has excellent positive inotropic effect, hypotensive effect, adenylate cyclase activating effect, and water solubility superior to that of forskolin. Therefore, the compound of the present invention is expected as a therapeutic drug for acute or chronic heart failure, an antihypertensive drug, and a cerebral vasodilator. In addition, it is expected as a therapeutic drug for diseases caused by dysregulation of cAMP such as glaucoma, asthma, immunodeficiency, tumors and digestive system diseases. The dose in this case is usually 0.01 to 30 mg / kg / day, although it varies depending on the symptoms, age and administration method of the patient to be administered.

In the present invention, the preferred compound is 6-dimethylaminoacetylforskolin (Compound No. 10).

6- (4-Dimethylaminobutyryl) forskolin (Compound No. 20)

7-Deacetyl-7- (2,3-dihydroxypropionyl) forskolin (Compound No. 27)

6- (4-aminobutyryl) forskolin (Compound No. 38)

6- (Pyrrolidinoacetyl) forskolin (Compound No. 39)

6- (4-dimethylaminobutyryl) -1
4,15-Dihydroforskolin (Compound No. 4
1 )

And the like. Of these, Compound No. is particularly superior in terms of drug efficacy . 2 0,38 and the like.

The compound of the present invention is mixed with an appropriate pharmaceutical carrier to prepare tablets, granules, fine granules, powders, capsules, injections, suppositories, eye drops, patches, ointments and the like, It is administered orally or parenterally. In particular, since it has a higher water solubility than that of forskolin, it can be made into an aqueous solution formulation suitable for infusion or intravenous administration, and therefore an excellent therapeutic effect is expected upon parenteral administration.

The compounds of the present invention will be described more specifically with reference to examples. Reference Example 1 7-Deacetyl-7-dimethylaminoacetylforskolin (Compound No. 1)

7-deacetylforskolin (500 m
g), pyridine (250 mg), dichloromethane (10
ml) mixed solution under ice cooling with chloroacetyl chloride (250
Dichloromethane solution (1 ml) is added and the mixture is stirred at room temperature for 1 hour. Then, under ice cooling, further pyridine (75m
g) and acetyl chloride (75 mg) in dichloromethane (1 ml) are added, and the mixture is further stirred for 3 hours at room temperature. After completion of the reaction, the reaction solution is concentrated under reduced pressure and dichloromethane and unreacted reagents are distilled off. 7-Deacetyl-7-chloroacetylforskolin was obtained as an oily substance, and the following reaction was carried out without purification. That is, this oily matter was converted into dichloromethane (1
It is dissolved in 0 ml), dimethylamine (3 ml) is added under ice cooling, and the mixture is stirred for 1 hour. After completion of the reaction, the reaction solution is concentrated under reduced pressure, water is added to the obtained residue, and the mixture is extracted with ethyl acetate. The organic layer is washed with water and dried over magnesium sulfate,
The desiccant is filtered off. The filtrate was concentrated under reduced pressure and the residue (78
0 mg) is purified by silica gel chromatography. By eluting with ethyl acetate, 7-deacetyl-
7-Dimethylaminoacetylforskolin (468m
(yield: 76% from g, 7-deacetylforskolin).

Mp 162-166 ° C. (hexane-ethyl acetate) IR (nujol) v: 3450, 3200, 173
5,1705 cm ^-1 MS m / z (relative intensity): 453 (M ⁺ , 5), 35
7 (6) 102 (71), 59 (52), 58 (10
0) The crystals are dissolved in dioxane, and equimolar hydrogen chloride dioxane solution is added to obtain a hydrochloride. mp 284 to 287 ° C (EtOH) IR (nujol) ν: 1740, 1710 cm ^-1

The following amine was used in place of the following amine in place of dimethylamine, and compound No. 2 was similarly used in the reaction with 7-deacetyl-7-chloroacetylforskolin.
~ 6 is obtained. Yield: Yield from 7-deacetylforskolin. IR and MS data are shown in Table 3.

[0071]

[Table 8]

[0072]

[Table 9]

Reference Example 2 7-Deacetyl-7- (3-dimethylaminopropionyl) forskolin (Compound No. 7) Forskolin (15 g), t-butyldimethylchlorosilane (11.25 g), imidazole (5.25).
A mixture of g) and N, N-dimethylformamide (45 ml) is stirred at 70 ° C. for 21 hours. After completion of the reaction, the reaction solution is poured into water and extracted with ethyl acetate. After washing the organic layer with water,
Dry over magnesium sulfate and filter off the desiccant. The filtrate is concentrated under reduced pressure to obtain crude 1- (t-butyldimethylsilyl) -forskolin (21.62 g).

This oil (21.62 g) was dissolved in methanol (250 ml), and 1N NaOH (4
0 ml) is added dropwise. Stir overnight at room temperature, and after the reaction is complete,
The reaction solution is concentrated under reduced pressure. Water is added to the obtained residue and the mixture is extracted with ethyl acetate. The organic layer is washed with water, dried over magnesium sulfate, and the desiccant is filtered off. The filtrate was concentrated under reduced pressure, and the residue (20 g) was purified by silica gel chromatography and eluted with hexane-ethyl acetate (5: 1) to give 1- (t-butyldimethylsilyl) -7-deacetyl as an oil. Forskolin (17.42 g, 99% from forskolin) is obtained.

IR (neat) ν: 3500, 3300, 1
710 cm ^-1 MS m / z (relative intensity): 482 (M ⁺ , 0.8),
466 (100), 407 (8), 311 (20), 1
91 (36), 75 (100)

This oil (1.5 g), pyridine (32
3-Chloropropionyl chloride (513 mg) is added dropwise to a mixed solution of 0 mg) and dichloromethane (10 ml). After stirring at room temperature for 5 hours, pyridine (320 mg) and β-chloropropionyl chloride (513 mg) were added. After stirring at room temperature overnight, after completion of the reaction, water is added and the mixture is extracted with ethyl acetate. The organic layer is washed with water, dried, and the desiccant is filtered off.
The filtrate was concentrated to give crude 1- (t-butyldimethylsilyl) -7- (3-chloropropionyl (-7
-Deacetylforskolin (2.09 g) is obtained.

To a solution of this oil (1 g) in dichloromethane (10 ml) was added an excess of dimethylamine under ice cooling. After stirring for 2 hours at room temperature, after completion of the reaction, the reaction solution was concentrated to give crude 1- (t-butyldimethylsilyl) -7-deacetyl-7- (3-dimethylaminopropionyl) forskolin as an oil (877 mg). To get

¹ H-NMR (CDCl ₃ ) δ: 5.44
(1H, d, J = 4.65 Hz, 4.62 (1H, br
s), 4.60 (1H, br s), 2.86 (6H,
s), 1.70 (3H, s), 1.45 (3H, s),
1.33 (3H, s), 1.26 (3H, s), 1.0
5 (3H, s), 0.87 (9H, s), 0.14 (3
H, s), 0.02 (3H, s).

A solution of this oil (865 mg) in methanol (10 ml) under ice cooling was added to trifluoroacetic acid (4 ml).
Is added and the mixture is stirred at room temperature for 43 hours. After completion of the reaction, the reaction solution is concentrated, the resulting residue is diluted with dilute hydrochloric acid, and washed with ethyl acetate. The aqueous layer is made alkaline with 28% aqueous ammonia and extracted with ethyl acetate. The organic layer is washed with water, dried over magnesium sulfate, and the desiccant is filtered off. Concentrate the filtrate,
The obtained residue is recrystallized from hexane-dichloromethane to obtain 7-deacetyl-7- (3-dimethylamino-propionyl) forskolin (283 mg). [1-
(41% from (t-butyldimethylsilyl) -7-deacetylforskolin]

Mp 150 = 153 ° C. MS m / z (relative intensity): 467 (M ⁺ , 2), 2
02 (2), 159 (8), 118 (29), 92 (6)
1), 91 (81), 58 (100). No. 2 using diethylamine instead of dimethylamine
The compound of 5 was obtained as a colorless oil. Yield 69.1% MS m / z: 495 (M ⁺ ).

Reference Example 3 7-deacetyl-7- (4-pyrrolidinobutyryl) forskolin (Compound No8) 7-deacetylforskolin (1.3 g), pyridine (700 mg), dichloromethane (40 ml) in a mixed solution under ice cooling, A dichloromethane solution (10 ml) of 4-chlorobutyryl chloride (800 mg) is added, and the mixture is stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution is concentrated under reduced pressure, and the resulting residue is purified by silica gel chromatography.
Elute with hexane-ethyl acetate to give 7- (4
-Chlorobutyryl) -7-deacetylforskolin (1.27g) is obtained.

¹ H-NMR (CDCl ₃ ) δ: 5.53
(1H, d, J = 5Hz), 4.58 (1H, br s),
4.48 (1H, br s), 3.64 (2H, t, J = 6H
z), 2.62 (2H, m), 2.18 (4H, m),
1.73 (3H, s), 1.45 (3H, s), 1.3
5 (3H, s), 1.27 (3H, s), 1.04 (3
H, s),

A mixture of this oil (1.27 g), pyrrolidine (20 ml) and dichloromethane (50 ml) was stirred at room temperature for 5 days. After the reaction is completed, the reaction solution is depressurized,
Concentrate. Water is added to the residue and extracted with ethyl acetate. The organic layer is washed with water, dried over magnesium sulfate, and the desiccant is filtered off. The filtrate is concentrated under reduced pressure and the residue is purified by silica gel chromatography. Elution with chloroform-methanol gave 7-deacetyl-7- (4-pyrrolidinobutilyl) phoruscholine (109 mg, 6% from 7-deacetylphoruscholine).

¹ H-NMR (CDCl ₃ ) δ: 5.37
(1H, d, J = 4Hz), 4.56 (1H, m),
4.50 (1H, m), 2.53 (8H, m), 1.9
3 (2H, m), 1.79 (4H, m), 1.73 (3
H, s), 1.44 (3H, s), 1.35 (3H,
s), 1.26 (3H, s), 1.03 (3H, s). MS m / z (relative intensity): 507 (M ⁺ , 4), 1
99 (3), 156 (26), 140 (10), 92
(40), 91 (52), 84 (100).

Reference Example 4 6-Dimethylaminoacetyl-7-deacetylforskolin (Compound No. 9) 7-deacetyl-7-dimethylaminoacetylforskolin (200 mg) obtained in Reference Example 1 was mixed with acetonitrile-water (45:55). It is dissolved in a solvent (20 ml), 1N aqueous sodium hydroxide solution (0.8 ml) is added, and the mixture is stirred at room temperature for 25 minutes. After completion of the reaction, the reaction solution is concentrated under reduced pressure. Water is added to the obtained residue, and the mixture is extracted with ethyl acetate. The organic layer is washed with water, dried over magnesium sulfate, and the desiccant is filtered off. The filtrate is concentrated under reduced pressure and the resulting residue (242 mg) is purified by silica gel chromatography. 6-by eluting with acetonitrile
Dimethylaminoacetyl-7-deacetylforskolin (176 mg, 88%) is obtained.

Mp 116-117 ° C. (hexane-ethyl acetate) IR (nujol) v: 3410,3200,175
0.1720 cm ^-1 MS m / z (relative intensity): 453 (M ⁺ , 5), 35
0 (2), 237 (2), 219 (2), 104 (1
5), 58 (100).

The crystals are dissolved in dioxane, and an equimolar hydrogen chloride-dioxane solution is added to obtain a hydrochloride. mp 263-265 ° C (EtOH) IR (nujol) v: 3490, 3230, 268
Using the corresponding 7-substituted forskolin at 0,1745,1710 cm ^-1 , the same transfer reaction, the compounds shown in Table 4 can be obtained.

[0088]

[Table 10]

[0089]

[Table 11]

[0090]

[Table 12]

Example 1 6-Dimethylaminoacetylforskolin (Compound N
o. 10) Pyridine (1 g) and acetyl chloride (750 mg) were added to a mixed solution of 6-dimethylaminoacetyl-7-deacetylforskolin (Compound No. 9, 587 mg) and dichloromethane (20 ml) in four portions. , Stir at room temperature for 7 hours. After completion of the reaction, water is added, the mixture is made basic with a saturated aqueous solution of sodium hydrogen carbonate and extracted with dichloromethane.
The organic layer is dried over magnesium sulfate and the desiccant is filtered off. The filtrate is concentrated and the resulting residue (811 mg) is purified by silica gel chromatography. Hexane-A
Elute with tell (1: 3) to give 6-dimethylaminoacetylforskolin (444 mg, 69%).

Mp 190-193 ° C. (toluene) IR (nujol) v: 3100, 1750, 173
^{0,1720cm -1 1 H-NMR (CDCl} 3) δ: 5.86 (1H, d,
J = 4 Hz), J = 2.7 Hz), 5.56 (1H,
d, J = 4.9 Hz), 4.61 (1H, br s), 3.
18 (2H, s), 2.31 (6H, s), 2.04
(3H, s), 1.64 (3H, s), 1.42 (3
H, s), 1.35 (3H, s), 1.04 (3H,
s), 0.96 (3H, s). The hydrochloride is obtained in the same manner as in Reference Example 4 . mp 255-260 ° C (EtOH) IR (nujol) v: 3240, 3130, 240
0,1750,1725cm ^-1

In this example, N was obtained by using propionyl chloride or butyryl chloride instead of acetyl chloride.
o. Compounds 11 and 12 are obtained (see Table 5).
Also, in this example, 6-dimethylaminoacetyl-7
6-diethylaminoacetyl-7-deacetylforskolin (No. 14
Compound), 6- (3-diethylaminopropionyl)
By using -7-deacetylforskolin (compound of No. 24) and 6- (4-dimethylaminobutyryl) -7-deacetylforskolin (compound of No. 19), the No. ． 15, 18, 20 compounds can be obtained. In addition, Reference Example compound No. 17 is also the same.
6- (3-Dimethylaminopropionyl) -7-deace
Tilforskolin (compound of No. 23) can be used
You can

[0094]

[Table 13]

[0095]

[Table 14]

[0096]

[Table 15]

[0097]

[Table 16]

[0098]

[Table 17]

[0099]

[Table 18]

Reference Example 5 : 6-Dimethylaminoacetyl-7-deacetyl-7-dimethylaminoacetylforskolin (Compound No. 1
3) 6-Dimethylaminoacetyl-7-deacetylforskolin (Compound No. 9, 700 mg), pyridine (14
Chloroacetyl chloride (209 mg) was added dropwise to a mixed solution of 6 mg) and dichloromethane (10 ml) under ice cooling.
The mixture is stirred under ice cooling for 3 hours, and after the reaction is completed, the reaction solution is concentrated. The residue obtained is diluted with water, made basic with saturated aqueous sodium hydrogen carbonate solution and extracted with ethyl acetate. The organic layer is washed with water, dried over magnesium sulfate, and the desiccant is filtered off. The filtrate is concentrated to give 7-chloroacetyl-7-deacetyl-6-dimethylaminoacetylforskolin (749 mg) as a white solid.

A dichloromethane solution (20 ml) of this white solid (749 g) was added dropwise to an excess dichloromethane solution of dimethylamine (20 ml) under water cooling, and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, water is added to the reaction solution and the mixture is extracted with dichloromethane. After drying the organic layer with magnesium sulfate, the desiccant is filtered off. The filtrate is concentrated and the resulting residue (728 mg) is purified by silica gel chromatography. Eluting with dichloromethane-acetone (2: 1),
6-Dimethylaminoacetyl-7-deacetyl-7-dimethylaminoacetylforskolin (186 mg, 23% from compound No. 9) is obtained. mp 180-183 ° C (hexane-ethyl acetate) MS m / z (relative intensity): 538 (M ⁺ , 58), 4
54 (8), 104 (100), 102 (100).

In a similar manner, 6-acetyl-7-deacetylforskolin was converted to 6-acetyl-7-chloroacetyl-7-deacetylforskolin and then reacted with dimethylamine to react with 6-acetyl-7. -Deacetyl-7-dimethylaminoacetylforskolin (Compound No. 21, 6-acetyl-7-deacetylforskolin, 88%) is obtained. ¹ H-NMR (CDCl ₃ ) δ: 5.82 (1 H, q,
J = 4.7 Hz, J = 3.0 Hz), 5.62 (1H,
d, J = 4.4 Hz), 4.61 (1H, br s), 3.
15 (2H, center of AB qnart-et. J = 17H
z), 2.37 (6H, s), 2.09 (3H, s),
1.66 (3H, s), 1.44 (3H, s), 1.3
3 (3H, s), 1.03 (3H, s), 0.98 (3
H, s).

In a similar manner, 6-acetyl-7-deacetylphoruscholine was treated with 4-chlorobutyryl chloride to give 6
Convert to -acetyl-7- (4-chlorobutyryl) 7-deacetylforskolin. ¹ H-NMR (CDCl ₃ ) δ: 5.83 (1 H, q,
J = 4.3 Hz, J = 3.1 Hz), 5.55 (1H,
d, J = 4.6 Hz), 4.63 (1H, br s), 3.
52 (2H, m), 2.38-2.6 (4H, m) 2.
10 (3H, s), 2.04-2.2 (3H, m),
1.673H, s), 1.44 (3H, s), 1.36
(3H, s), 1.04 (3H, s), 0.98 (3
H, s).

This compound was reacted with pyrrolidine to give 6-acetyl-7-deacetyl-7- (4-pyrrolidinobutilyl) forskolin (Compound No. 22, 6-acetyl-7-deacetylforskolin 14 %). ¹ H-NMR (CDCl ₃ ) δ: 5.81 (1H, q,
J = 4.3 Hz, J = 2.9 Hz), 5.54 (1H,
d, J = 4.6 Hz), 4.60 (1H, br s), 2.
62 (6H, m), 2.28-2.44 (3Hm),
2.09 (3H, s), 1.75-1.95 (6H,
m), 1.65 (3H), 1.44 (3H, s), 1.
35 (3H, s), 1.03 (3H, s), 0.98
(3H, s).

Reference Example 6 7-Deacetyl-7-hemiglutamine forskolin (Compound No. 26) A mixture of 7-deacetylforskolin (369 mg), glutamic anhydride (342 mg) and pyridine (5 ml) is heated at 110 ° C. for 2 hours. After completion of the reaction, the reaction solution is concentrated under reduced pressure, water is added to the obtained residue, and the mixture is extracted with ethyl acetate. The organic layer is washed with an aqueous solution of copper sulfate, dried over magnesium sulfate, and then the desiccant is filtered off. The filtrate is concentrated under reduced pressure and the resulting residue is purified by silica gel chromatography. Elution with hexane-acetic acid (2: 1) then chloroform-methanol (9: 1) gave 7-deacetyl-7-hemiglutarylphoscholine (Compound No. 26, 270 mg, 56%) as an oil.

¹ H-NMR (CDCl ₃ ) δ: 5.49
(1H, q, J = 4.0Hz), 4.58 (1H, br,
s), 4.47 (1H, t.J = 3.7 Hz), 2.5
0 (4H, m), 2.20 (2H, m), 2.05 (2
H, m), 1.73 (3H, s), 1.45 (3H,
s), 1.35 (3H, s), 1.27 (3H, s),
1.04 (3H, s). MS m / z (relative intensity): 482 (M ⁺ , 7), 46.
4 (62), 436 (10), 227 (41), 115
(84), 81 (100)

Example 2 7-Deacetyl-7- (2,3-dihydroxypropionyl) forskolin (Compound No. 27) 7-deacetylforskolin (500 mg), pyridine (300 mg), dichloromethane (10 ml) in a mixed solution under ice cooling, 2 7 by eluting with 2-dimethyl-1.3-diodichloromethane-ethyl acetate (2: 1)
-Diacetyl-7- (2,3-dihydroxypropionyl) forskolin diastereomeric mixture (A: B
= 3: 2). (250 mg, 91%) mp 160-172 ° C IR (nujol) ν: 3440, 3320, 1750, 17
30,1700 cm- ⁺ MS m / z (relative intensity): 456 (M ⁺ , 5), 43
8 (77), 123 (94), 99 (80), 81 (1
00).

Example 3 6-hemisuccinylforskolin (Compound No. 29) (1) Forskolin (1 g), t-butyldimethylsilyl chloride (1.2 g), imidazole (1.25)
A mixture of g) and N, N-dimethylformamide (2 ml) is left for 2 hours at 60 ° C. The reaction solution is poured into ice water and extracted with ether. The ether layer is washed with water and dried over anhydrous sodium sulfate. After the drying agent is filtered off, the filtrate is concentrated under reduced pressure, a dichloromethane solution (4 ml) of quinlan-4-carboyl chloride (360 mg) is added, and the mixture is stirred at room temperature for 4 hours. After completion of the reaction, the reaction solution is concentrated under reduced pressure, and the obtained residue (950 mg) is purified by silica gel chromatography. Dichloromethane-ethyl acetate (10:
By elution with 1), 7-deacetylforskolin-7- (2,2-dimethyl-1,3-dioxolane-
A diastereomeric mixture of 4-carboxylates is obtained. (557 mg, 83%) mp 160-165 ° C MS m / z (relative intensity): 496 (M ⁺ , 2), 47.
8 (40), 131 (32), 123 (36), 101
(100), 59 (30),

This 7-deacetylforskolin-7-
(2,2-Dimethyl-1,3-dioxolane-4-carboxylate (300 mg), 70% acetic acid (10 ml)
The mixture is stirred at 60 ° C. for 2 hours. After completion of the reaction, the reaction solution is concentrated under reduced pressure, and the obtained residue (470 mg) is purified by silica gel chromatography. 1- (t-Butyldimethylsilyl) forskolin (1.2 g) is obtained as a colorless oil.

(2) The obtained oily substance (1.2 g) is dissolved in methanol (10 ml), 1N NaOH is added, and the mixture is stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution is diluted with water and extracted with ethyl acetate. The organic layer is washed with water, dried over anhydrous sodium sulfate, and the desiccant is filtered off. The filtrate is concentrated under reduced pressure, and the residue (1.0 g) is purified by silica gel chromatography. Elution with hexane-ethyl acetate (5: 1) gave 1- (t-butyldimethylsilyl) -7-deacetylforskolin (0.85 g) as a colorless oil.

(3) 1- (t-butyldimethylsilyl)
After dissolving -7-deacetylforskolin (0.44 g) in pyridine (2.0 ml), succinic anhydride (4 g) is added and the mixture is refluxed for 2 hours. After completion of the reaction, pyridine is concentrated under reduced pressure, and the residue is thoroughly washed with ethyl acetate and 0.1N HCl for extraction. The organic layer is washed with water and dried over anhydrous sodium sulfate. The desiccant was filtered off, the filtrate was concentrated under reduced pressure, and
(T-Butyldimethylsilyl) -7-hemisuccinyl-
7-deacetylforskolin (0.53 g, quantitative)
As an oil.

(4) This oil (250 mg) was dissolved in a mixed solvent (10 ml) of acetonitrile-water (1: 1), 1N aqueous sodium hydroxide solution (1 ml) was added, and the mixture was stirred at room temperature for 30 minutes. After completion of the reaction, dilute the reaction solution with water,
The pH is adjusted to 3 with 0.5N HCl and then extracted with ethyl acetate. The organic layer is washed with water and dried over anhydrous sodium sulfate.
The desiccant is filtered off, the filtrate is concentrated, and the resulting residue is purified by silica gel column chromatography. 1- (t-
Butyldimethylsilyl) -6-hemisuccinyl-7-deacetylforskolin is a colorless oil (178 mg, 7
1.2%).

(5) This oily substance (178 mg) was dissolved in pyridine (0.4 ml), and acetic anhydride (0.4 m
l) Add and leave overnight at 4 ° C. The reaction solution is poured into ice water and extracted with ethyl acetate. The organic layer is washed with water and then dried over anhydrous sodium sulfate. After the desiccant is filtered off, the filtrate is concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to give 1- (t-butyldimethylsilyl) -6-hemisuccinylforskolin (183 m).
g, 96.3%) as a colorless oil.

(6) This oil (75 mg) was dissolved in methanol (1 ml), and trifluoroacetic acid (1.5 m) was added.
1) is added and the mixture is stirred at room temperature for 4 hours. Saturated NaHC in the reaction solution
An O ₃ aqueous solution is added to neutralize, and the mixture is extracted with ethyl acetate. After washing the organic layer with water, the filtrate was concentrated under reduced pressure to give 6-hemisuccinyl-forskolin (Compound No. 29, 32 m
g, 52.2%) as a colorless oil. NMR (CDCl ₃ ) δ: 1.01, 1.38, 1.4
2, 1.61, 1.69, 2.01 (each, 3H, s, M
e), 2.01 (3H, s, OAc), 2.60-2.
80 (4H, m, -COC H 2 C H 2 CO -), 4.6
1 (1H, broad, s, 1β-H), 4.98 (1H,
d, J = 8 Hz, 15− H ), 5.28 (1 H, d, J
= 17.5 Hz, 15- H ), 5.52 (1H, d, J
= 3 Hz, 72- H ), 5.83 (1H, t, 3.5H
z, 62-H), 5.97 (1H, dd, J = 8, 1,
7.5 Hz, 14- H )

Reference Example 7 : 6-hemisuccinyl-7-deacetylforskolin (Compound No. 28) 1- (t-butyldimethylsilyl) -6-hemisuccinyl-7-deacetylforskolin (260 mg) obtained in Example 3 (4) ) Is deprotected in the same manner as in Example 3 (6) to obtain 6-hemisuccinyl-7-deacetylforskolin (Compound 28, 140 mg, 67%) as a colorless oil. MS m / z (relative intensity): 468 (M ⁺ ).

Example 4 Forskolin-1-hemisuccinate (Compound No. 3
1) Forskolin (1 g), succinic anhydride (4.9 g),
A mixture of 4-morpholino-N, N'-dicyclohexylcarbodiimide (1.1 g) and pyridine (8 ml) is heated at 100 ° C for 1 hour. After completion of the reaction, the reaction solution is concentrated under reduced pressure, 0.5N hydrochloric acid is added to the obtained residue, and the mixture is extracted with ethyl acetate. The ethyl acetate layer is washed with saturated brine, dried over anhydrous sodium sulfate, and the desiccant is filtered off.
The filtrate is concentrated under reduced pressure, and the resulting residue is purified by silica gel column chromatography. Chloroform-methanol (96: 4), then chloroform-methanol-
Forskolin-1-hemisuccinate (Compound No. 31, 0.7 g) eluted with acetic acid (18: 2: 0.5).
As a colorless oil.

¹ H-NMR (sodium salt, CDC
l ₃ ) δ: 5.50 (1H, d, J = 4 Hz), 5.5
0 (1H, s), 4.46 (1H, br s), 2.50
~ 2.30 (4H, m), 2.16 (3H, s), 1.
67 (3H, s), 1.51 (3H, s), 1.31
(3H, s), 1.25 (3H, s), 1.02 (3
H, s). MS (EI.70 eV. Methyl ester derivative) m /
z (relative intensity): 506 ([M-18] ⁺ , 5), 24
3 (7), 191 (33), 151 (31), 115
(100), 107 (45).

The sodium salt of forskolin-1-hemisuccinate is obtained by dissolving it in a 5% aqueous sodium hydrogen carbonate solution, adsorbing it on Amberlite XAD-II, washing it with water, and eluting with methanol.

In the same manner, forskolin-1-hemiglutarate (Compound No. 32) can be produced by using glutaric anhydride instead of succinic anhydride. ¹ H-NMR (sodium salt, CDCl ₃ ) δ: 5.5
7 (1H, br s), 5.48 (1H, d, J = 4H
z), 4.55 (1H, t, J = 3.7Hz), 2.5
~ 2.2 (4H, m), 2.0 to 1.8 (2H, m),
2.18 (3H, s), 1.71 (3H, s), 1.5
4 (3H, s), 1.34 (3H, s), 1.28 (3
H, s), 1.05 (3H, s). MS (EI.70 eV. Methyl ester derivative) m /
z (relative intensity): 520 ([M-18] ⁺ 30), 25
9 (7), 219 (5), 191 (3), 175
(6), 129 (100).

Example 5 Forskolin-1-Hemiadipate (Compound No. 3
3) Pyridine (0.2 ml) was added to a dichloromethane solution (10 ml) of forskolin (0.12 g) and adipic dichloride (0.5 g) under ice cooling, and the reaction solution was stirred at room temperature for 4 hours.
Stir for hours. The reaction solution was poured into an ice-cooled 5% aqueous sodium hydrogen carbonate solution to hydrolyze the acid chloride and
Adjust pH to 3 with 5N hydrochloric acid. Ethyl acetate is added to this and extraction is performed. The organic layer is washed with saturated saline and then dried over anhydrous sodium sulfate. The extract is concentrated under reduced pressure,
The residue obtained is purified by silica gel thin layer chromatography. Chloroform-methanol acetic acid (9: 1: 0.
The solution was developed in 1) and the portion having a pH value of 0.3 to 0.5 was fractionated to obtain forskolin-1-hemiadipate (Compound No. 33, 0.065 g) as a colorless oil.

¹ H-NMR (sodium salt, CDC
l ₃ ) δ: 5.56 (1H, br s), 5.51 (1
H, d, J = 4 Hz), 4.46 (1H, t, J = 3.
2 Hz), 2.5 to 2.0 (4 H, m), 1.8 to 1.
5 (4H, m), 2.18 (3H, s), 1.70 (3
H, s), 1.53 (3H, s), 1.33 (3H,
s), 1.24 (3H, s), 1.05 (3H, s). MS (EI.70 eV. Methyl ester derivative) m /
z (relative intensity): 534 ([M-18] ⁺ 68), 28
2 (7), 259 (14), 219 (13), 191
(61), 143 (100). Then, a sodium salt is prepared in the same manner as in the case of compound No. 31.

Forskolin-1-hemioxalate (Compound No. 30) can be produced in the same manner. MS (FAB) m / z: 483 (M + H), 591 (M
H + 108).

Reference Example 8 : Forskolin-1- (N, N-dimethylaminobutyrate (Compound No. 36) Forskolin (0.20 g), N, N-dimethylaminobutyrate (0.25 g), dicyclohexylcarbodiimide (0.40 g), A mixture of 4-dimethylaminopyridine (0.30 g) and dichloromethane (5 ml) is reacted at room temperature for 2 hours, the precipitated crystals are filtered off, and the filtrate is made alkaline with 5% aqueous sodium hydrogen carbonate solution, Extract with ethyl acetate, wash the organic layer with water,
Dry over anhydrous sodium sulfate. The desiccant is filtered off, the filtrate is concentrated under reduced pressure, and the resulting residue is purified by silica gel chromatography. Elution with ethyl acetate and then ethyl acetate-methanol-triethylamine (95: 5: 1) gave forskolin-1- (N, N-dimethylaminobutyrate) (Compound No. 36, 0.19 g) as a colorless oil. Get as a thing.

¹ H-NMR (CDCl ₃ ) δ: 5.55
(1H, s), 5.53 (1H, d, J = 4.8H
z), 4.47 (1H, br s), 2.5 to 2.1 (4
H, m), 2.18 (6H, s), 2.17 (3H,
s), 1.71 (3H, s), 1.85 to 1.65 (2
H, m), 1.54 (3H, s), 1.35 (3H,
s), 1.28 (3H, s), 1.06 (3H, s) MS (FAB) m / z (relative intensity): 524 (M + H,
100), 632 (MH + 108, 78).

Reference Example 9 Forskolin-1-glycinate (Compound No. 34) (1) Forskolin (0.3 g), t-butoxycarbonylglycine (0.3 g), dicyclohexylcarbodiimide (0.4 g), 4-dimethylaminopyridine ( A mixture of 0.3 g) and dichloromethane (5 ml) is stirred at room temperature for 2 hours. After separating the precipitated crystals by filtration, the filtrate is extracted with ethyl acetate. The organic layer is washed with water and then dried over anhydrous sodium sulfate. After the desiccant is filtered off, the filtrate is concentrated under reduced pressure and the residue is subjected to silica gel column chromatography. Elute with ethyl acetate-benzene (5:95), then ethyl acetate-benzene (10:90),
Forskolin-1-BOC-glycinate (0.31
g) is obtained as a colorless oil.

¹ H-NMR (CDCl ₃ ) δ: 5.61
(1H, br s), 5.42 (1H, d, J = 4H
z), 4.45 (1H, s), 3.80 (2H, d, J
= 5.3 Hz), 2.17 (3H, s). 1.74 (3
H, s), 1.55 (3H, s), 1.45 (9H,
s), 1.34 (3H, s), 1.29 (3H, s),
1.05 (3H, s).

(2) Next, this forskolin-1-B
OC-glycineate (0.31 g) was mixed with acetic acid (0.5 m
l) and dissolved under ice cooling in a hydrobromic acid-acetic acid solution (0.5
ml) is added and the mixture is allowed to stand at room temperature for 10 minutes. After neutralizing the excess hydrobromic acid with 5% sodium hydrogen carbonate, it is adsorbed on Amberlite XAD-2 resin. The column was washed with water and then eluted with MeOH to obtain forskolin-1-glycineate (Compound No. 34, 0.12 g) as a colorless oil. MS (FAB) m / z (relative intensity): 468 (MH),
576 (MH + 108).

Forskolin-1 was prepared by the same method.
-(6-aminocaproate) (Compound No. 37), forskolin-1- (4-aminobutyrate) (Compound
No. 35) can be manufactured. Forskolin-1- (6-aminocaproate) (Compound No. 37) oil MS (FAB) m / z: 524 (M + H), 632 (M
H + 108). Forskolin-1- (4-aminobutylate) (Compound No. 35) oil MS (FAB) m / z: 496 (M + H), 604 (M
H + 108).

Example 6 6- (4-aminobutyryl) forskolin (Compound N
o. 38) (1) 7-deacetylforskolin (3.00 g),
To a mixture of dichloromethane (100 ml) under ice cooling,
4- (t-butoxycarbonylamino) butyric acid (8.20
g), 4-dimethylaminopyridine (6.40 g) and dicyclohexylcarbodiimide (12.60 g) are added in 4 portions, and the mixture is stirred at room temperature overnight. After completion of the reaction, water (0.5 ml) is added and the reaction solution is concentrated under reduced pressure. Dichloromethane is added to the concentrated solution, and the lower solution is filtered off. The filtrate is concentrated, and the obtained residue is purified by silica gel column chromatography. Dichloromethane-ethyl acetate (1
Elution at 0: 1) gave 7- [4- (t-butoxycarbonylamino) butyryl] -7-deacetylforskolin as an oil (Compound No. 38a: 1.60 g, 36%).
To get MS m / z (relative intensity): 535 [MH ₂ O] ⁺ ,
23), 479 (23), 104 (34), 86 (8)
1), 57 (100).

(2) This oily substance (No. 38a, 1.50)
g), acetonitrile (90 ml) and water (60 ml) were added to a 1N aqueous sodium hydroxide solution (6 ml), and the mixture was stirred at room temperature for 40 minutes. After completion of the reaction, the reaction solution is immediately concentrated, water is added, and the mixture is extracted with ethyl acetate. The extract is dried over magnesium sulfate, the desiccant is filtered off, and the filtrate is concentrated. The resulting residue (1.43 g) is purified by silica gel column chromatography. Elute with dichloromethane-ethyl acetate (4: 1 to 2: 1) to give 6- [4- (t
-Butoxycarbonylamino) butyryl] -7-deacetyl-forskolin (Compound No. 38b, 1.12
g, 75%). mp 188-190 ° C (ethyl acetate) IR (nujol) ν: 3350, 1735, 1710
cm ^-1

(3) This compound (No. 38b: 1.12)
g) was dissolved in dichloromethane (80 ml), and pyridine (1.52 g) and acetyl chloride (675 mg) were dissolved in this solution.
Is added in 3 portions. After stirring at room temperature for 8 hours, water is added to complete the reaction. Extract with dichloromethane and dry the organic layer over magnesium sulfate. The desiccant is filtered off, the filtrate is concentrated, and the resulting residue (1.97 g) is purified by silica gel column chromatography. Elute with dichloromethane-ethyl acetate (10: 1 to 5: 1) to give 6- [4- (t
-Butoxycarbonylamino) -butyryl] forskolin (Compound No. 38c: 414 mg, 34%) is obtained. Compound No. 38c: mp 162-165 ° C. (hexane-
toluene)

(4) This compound (No. 38c, 123m
g) is dissolved in 85% formic acid under ice cooling, and the mixture is stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution is concentrated, water is added and the mixture is washed with ethyl acetate. The aqueous layer is made basic with 3N aqueous sodium carbonate solution and extracted with ethyl acetate. The organic layer is dried over magnesium sulfate and the desiccant is filtered off. The filtrate was concentrated, and the obtained residue was recrystallized from ethyl acetate-methanol to obtain 6- (4-aminobutyryl) forskolin (Compound No. 38, 31 mg, 30%). mp 191-192 ° C. IR (Nujo - ^{Le) ν: 3360,1725cm -1 1 H-} NMR (CD 3 OD) δ: 5.76 (1H, q,
J = 4.7 Hz, J = 3.3 Hz), 5.39 (1H,
d, J = 4.3 Hz), 4.42 (1H, brs), 2.
59 (2H, t, J = 6.0 Hz), 2.2-2.4.
(2H, m), 1.88 (3H, s), 1.8 to 1.6
(3H, m), 1.56 (3H, s), 1.37 (3
H, s), 1.23 (3H, s), 0.91 (6H,
s). MS m / z (relative intensity): 495 (M ⁺ , 14), 1
46 (32), 104 (100), 99 (13), 86
(74).

Example 7 6-Pyrrolidinoacetylforskolin (Compound No. 3
9) (1) Example 6 7-deacetyl-7-pyrrolid obtained in the same manner as in Example 6 (1) except that pyrrolidinoacetic acid was used in place of 4- (t-butoxycarbonylamino) butyric acid in (1). Dinoacetylforskolin (4.20
g) is a mixed solvent of acetonitrile-water (4: 1) (15)
0 ml) and 1N aqueous sodium hydroxide solution (9.63 ml) was added dropwise under ice cooling. The mixture is stirred at room temperature for 6 hours, and water is added to terminate the reaction. The reaction solution is extracted with ethyl acetate and the organic layer is concentrated. The concentrate is dissolved in ether and extracted with 1N hydrochloric acid. Aqueous ammonia is added dropwise to the aqueous layer under ice cooling to make it basic, and the mixture is extracted with ether. The organic layer is dried over magnesium sulfate and the desiccant is filtered off. The filtrate is concentrated, and the obtained residue is purified by silica gel column chromatography. Elution with dichloromethane-methanol (20: 1) gave 7-deacetyl-6-pyrrolidinoacetylforskolin (Compound No. 39a, 900 mg, 21%). IR (KBr) ν: 3460, 3210, 1745,
1710 cm ^-1 MS m / z (relative intensity): 479 (M ⁺ , 1), 13
0 (8), 84 (100).

(2) This compound (No. 39a, 100m
g) and dichloromethane (10 ml) under ice-cooling, pyridine (72 mg), acetyl chloride (72 mg)
Is added in 4 portions over 6 hours, each added. Stir at room temperature overnight and extract with dichloromethane. The organic layer is washed with saturated saline and dried over magnesium sulfate. The desiccant is filtered off, and the residue (120 mg) obtained by concentrating the filtrate is purified by silica gel column chromatography. Chloroform-methanol (20: 1) was used to elute 6-pyrrolidinoacetylforskolin (Compound No. 39, 53).
mg, 50%). mp 166-168 ° C (methylene chloride-hexane) IR (KBr) ν: 3450, 3230, 1750, 1
715 cm ^-1

Reference Example 10 7-deacetyl-7- (4-dimethylaminobutyryl)
Forskolin (Compound No. 40) Of 7-deacetylforskolin (8.00 g), 4-dimethylaminobutyric acid / hydrochloride (14.56 g), 4-dimethylaminopyridine (21.21 g), dichloromethane (150 ml) Dicyclohexylcarbodiimide (35.82 g) and dichloromethane (1
20 ml) is added dropwise and the mixture is stirred at room temperature for 4 hours.
After completion of the reaction, the reaction solution is extracted with dilute hydrochloric acid. The aqueous layer is made basic with concentrated aqueous ammonia and extracted with diethyl ether.
The organic layer is washed with water and dried over anhydrous magnesium sulfate. The desiccant was filtered off, and the obtained filtrate was concentrated under reduced pressure to give 7-deacetyl-7- (4-dimethylaminobutyryl) forskolin (5.41 g, 52% from 7-deacetylforskolin) as an oil. %). MS m / z: 484 (M ⁺ ).

Example 8 6- (4-Dimethylaminobutyryl) -14,15-dihydroforuscholine (Compound No. 41) 6- (4-Dimethylaminobutyryl) forskolin (30 mg), 5% palladium-carbonic acid catalyst (6 mg),
A mixture of methanol (8 mg) is stirred under a hydrogen atmosphere at room temperature for 5 hours. After completion of the reaction, the catalyst was filtered off and the obtained filtrate was concentrated to give 6- (4-dimethylaminobutyryl)-
14,15-Dihydro-forskolin (Compound No. 4
1, 30 mg, 100%). mp 210-212 ° C (methanol) IR (nujol) ν: 3160, 1730, 1710
cm ^-1 (NMR and MS data are shown in Table 7) N as shown in Tables 6 and 7 in the same manner as in this example.
o. 42 to 45 compounds are obtained.

[0137]

[Table 19] The starting material of Reference Example compound No. 42 was the same as in Reference Example 10 in place of 4-dimethylaminobutyric acid / hydrochloride.
Other than that, N, N-dimethylglycine hydrochloride can be used to obtain the same. In addition, the starting materials for the compounds of Nos. 43, 44, and 45 were 4- (t-) in Example 6 (1).
Butoxycarbonylamino) butyric acid instead of N, N-
Dimethylglycine / hydrochloride, pyrrolidinoacetic acid or 3-dimethylaminopropionic acid may be used, and otherwise the same can be obtained as in Example 6 .

Example 9 13-cyclopropyl-6-dimethylaminoacetyl-
14,15-Dinorforskolin (Compound No. 46) 6-Dimethylaminoacetylforskolin (100 m
g), palladium acetate (14 mg) and anhydrous tetrahydrofuran (5 ml) are added to an ether solution of diazomethane under ice cooling, and the mixture is further stirred under ice cooling for 3 hours. After the reaction was completed, water was added and the pH was adjusted to 1 with 3N sodium carbonate
Set to 0 and extract with ethyl acetate. The organic layer is dried over magnesium sulfate and the desiccant is filtered off. The obtained filtrate is concentrated, and the residue is purified by silica gel column chromatography. Elution with dichloromethane-ether (4: 1 to 2: 1) gave 13-cyclopropyl-6-dimethylaminoacetyl-14,15-dinorforskolin (compound N
o. 46, 16 mg, 16%). IR (nujol) v: 3570, 3180, 173
^{5,1715cm -1 1 H-NMR (CDCl} 3) δ: 0.30 (2H,
m), 0.41 (1H, m), 0.75 (1H, m),
0.91 (1H, m), 0.95 (3H, s), 1.0
3 (3H, s), 1.32 (3H, s), 1.44 (3
H, s), 1.54 (3H, s), 2.01 (3H,
s), 2.43 (6H, s), 3.23 (2H, s),
4.51 (1H, br s), 5.51 (1H, d, J = 5
Hz), 5.83 (1H, t, J = 4Hz). MS m / z (relative intensity): 510 (M ⁺ , 23), 1
09 (49), 104 (100), 81 (49), 59
(100), 58 (100),

In a similar manner, 13-cyclopropyl-7-deacetyl-7-dimethylaminoacetyl- was obtained from 7-deacetyl-7-dimethylaminoacetylforskolin.
14,15-Dinorforskolin (Compound No. 47,
60%). IR (nujol) v: 3250, 1740, 1705
^{cm -1 1 H-NMR (CDCl} 3) δ: 0.18 (2H,
m), 0.49 (1H, m), 0.89 (1H, m),
0.96 (1H, m), 1.01 (3H, s), 1.2
8 (3H, s), 1.30 (3H, s), 1.50 (3
H, s), 1.66 (3H, s), 2.38 (6H,
s), 3.27 (2H, s), 4.43 (1H, br)
s), 4.47 (1H, br s), 5.43 (1H, d,
J = 4.5 Hz). MS m / z (relative intensity): 467 (M ⁺ , 4), 10
9 (26), 104 (58), 102 (60), 85
(41), 58 (100).

Reference Example 11 7-Deacetyl-7- (3-dimethylaminopropionyl) forskolin (Compound No. 7) 4-Dimethylaminopyridine (3.00 g) in a dichloromethane solution (60 ml) of 7-deacetylforskolin (2.00 g), 3-Dimethylaminopropionic acid
Hydrochloride (3.80 g) and dicyclohexylcarbodiimide (7.00 g) are added in 3 portions over 47 hours, and the mixture is stirred at room temperature. After completion of the reaction, the solvent is concentrated under reduced pressure, ethyl acetate is added to the obtained residue, and the mixture is filtered. Concentrate the filtrate,
The residue obtained (6.05 g) is purified by silica gel chromatography. Dichloromethane-methanol (2
Elution with 0: 1 to 10: 1) and 7-deacetyl-7-
(3-Dimethylaminopropionyl) forskolin (Compound No. 7, 1.72 g, 68%) is obtained.

[0141]

[Table 20]

[0142]

[Table 21]

Continuation of front page (51) Int.Cl. ⁶ Identification number Office reference number FI technical display location A61K 31/35 AED examiner Shinichi Naito (56) References JP-A-61-210081 (JP, A)

Claims (1)

[Claims] 1. A compound represented by the general formula ( I ′ ): [In the formula, I. When R ¹ is a hydrogen atom and R ⁴ is a vinyl group, an ethyl group or a cyclopropyl group, (1) R ² is a partial structural formula CO (CH ₂ ) mNR ⁵ R ⁶ (wherein R ⁵ and R ⁶ are hydrogen. Atom, lower alkyl group or R
⁵ is a lower alkylene group in which R ⁶ is bonded and an oxygen atom or a nitrogen atom may be contained in the bonding chain thereof, and m is 1 to
5 is an integer of 5), and R ³ is a lower alkylcarbonyl group.
Le group (R ³ is an acetyl group, R ² when R ⁴ is a vinyl group is a group other than dimethylamino propionyl group.) Or shown or, (2) R ² is a hydrogen atom, R ³ is -COCH (O
H) CH ₂ OH or (3) R ² represents —COCH ₂ CH ₂ CO ₂ H and R ³
Is _{-COCH 3, -COCH 2 CH 2 CH} 2 CO 2 H,
Or II showing a -COCH (OH) CH ₂ OH. When R ¹ represents a group represented by the partial structural formula CO (CH ₂ ) pCO ₂ H (where p is an integer of 0 to 5), R ²
Is a hydrogen atom, R ³ is an acetyl group, and R ⁴ is a vinyl group], and a physiologically acceptable salt thereof.