AU598724B2 - Triterpenyl esters of organic acids, process for their production, and hypolipidemic agents composed of them - Google Patents

Description

I FORM 10 598724 SPRUSON FERGUSON COMMONWEALTH OF AUSTRALIA PATENTS ACT 1952 COMPLETE SPECIFICATION

(ORIGINAL)

FOR OFFICE USE: Class Int. Class 0000 aoo o0 o0 0 0 0 0 be b en0 o~ at* Complete Specification Lodged: Accepted: Published: Priority: Related Art: ~Hy777772. :ci t i Name of Applicant: Address of Applicant: Actual Inventor(s): 0 000 0 0 00 *r 0 *r p 0000 pr AMANO PHARMACEUTICAL CO., LTD.

2-7, Nishiki 1-chome, Naka-ku, Nagoya-shi, Aichi-ken, Japan GORO KIMURA, YOSHIHIKO HIROSE, KUMI YOSHIDA, FUMIO KUZUYA and KATSUNARI FUJITA Spruson Ferguson, Patent Attorneys, Level 33 St Martins Tower, 31 Market Street, Sydney, New South Wales, 2000, Australia t~- Address for Service: Complete Specification for the invention entitled: "TRITERPENYL ESTERS OF ORGANIC ACIDS, PROCESS FOR THEIR PRODUCTION, AND HYPOLIPIDEMIC AGENTS COMPOSED OF THEM" The following statement is a full description of this invention, including the best method of performing it known to us SBR/JS/0238W

I

1 ABSTRACT OF THE DISCLOSURE A triterpenyl ester of organic acid other than triterpenyl esters of ferulic acid and of monobasic and dibasic saturated fatty acids. A process for producing a triterpenyl ester of organic acid other than esters of ferulic acid and of monobasic and dibasic saturated fatty acids, which comprises the reaction of a triterpenyl alcohol with an acid halide of the corresponding organic acid. A pharmaceutical composition for treatment of 10 hyperlipidemia comprising a pharmaceutical carrier and an effective amount of a triterpenyl ester of organic acid other than triterpenyl esters of dibasic saturated fatty o" acids. A pharmaceutical composition for treating hyperli- O o pidemia which comprises a pharmaceutical carrier and an effective amount of cyclobranol as an active ingredient.

02 o o A o2 02 a -<jS I-i i 1 TITLE OF THE INVENTION Triterpenyl Esters of Organic"Acids,Process for Their Production, and Hypolipidemic Agents Composed of Them BACKGROUND OF THE INVENTION Field of the Invention I I I. 4** I 0* i0 Tn )O The present invention relates to novel compounds having anti-hyperlipidemic activity and to processes for their production, and more particularly, is converned with novel triterpenyl esters of organic acids [except triterpenyl esters of ferulic acid (4-hydroxy-3-methoxycinnamic acid) and of monobasic and dibasic saturated fatty acids] 15 having excellent anti-hyperlipidemic activity and low toxicity and with processes for the production of these esters.

The invention further relates to antiatherosclerotic and hypolipidemic agents which contain the above-mentioned novel compounds and the known compounds cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of ferulic acid, or monobasic saturated fatty acids, and cyclobranol).

More specifically, the present invention relates to excellently active and low toxic agents for treatment of hyperlipidemia or hyperlipoproteinemia, that is, safety and novel hypolipidemic and r P ft tr *ttf I, C It 2 1 antiatherosclerotic agents which contain triterpenyl esters of organic acids, preferably, cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of organic acids including esters from triterpenyl alcohols and ferulic acid or monobasic saturated

C

4

C

20 fatty acids, and cyclobranol alone, as respective active ingredients.

Description of the Prior Art It is well known that hyperlipidemia or hyperlipoproteinemia is one of the most serious factors causing atherosclerosic, a form of arteriosclerosis, especially coronary heart disease.

Miller and Miller Miller and N.E. Miller, Lancet Jan. 4, p. 16 (1975)) have observed a negative correlation between the concentration of high density lipoprotein choresterol (hereinafter referred to as HDL-C) in blood plasma and the cholesterol pool in the body, and no correlation between either the concentration of total cholesterol (hereinafter referred to as TC) or the concentration of other lipoproteins, and the cholesterol pool, and therefrom proposed the theory that the reduction of cholesterol clearance from the arteries caused by a decrease in the HDL-C concentration in blood promotes arteriosclerosis. Since the proposal of this theory, a number of epidemiological studies T. Gordon et al., Am. J. Med., 62. 707 (1977)) have proved CIi 3 1 the presence of a reverse correlation between the onset of ischemic heart- disease and the concentration of HDL-C, and comfirmed that a decrease in the HDL-C concentration in blood is one of the most serious factors causing ischemic heart disease regardless of the presence or absence of an antihyperlipidemic agent.

It has so far been known that phytosterols reduce the content of cholesterol in serum. For 10 example, a mixture of 3-sitosterol and dihydro-3- 1sitosterol (supplied by Lilly Co., U.S.A. under the tradename of Cytellin) and a mixture of soysterol, phytosterol, and tocopherol (supplied by Morishita o Pharaceutical Co., Ltd. Japan, under the trandename of Moristerol) are on the market as anti-hyperlipedemic agents.

Q 06 I° On the other hand, the following is reported 0 o on triterpenyl alcohols.

Japanese Patent Application Laid-Open No.

18617/1982 describes that when a phytosterol (1 part) was used jointly with a cycloartenol or 24methylenecycloartanol (0.01 0.1 part), a stronger action of lowering serum cholesterol than that of phytosterol singly was exhibited by synergism.

Japanese Patent Application Laid-Open No.

116415/1983 also describes that a considerably stronger action depressing serum cholesteroldue to 4 1 synergistic effect was observed in the case of joint use of a phytosterol (100 parts) with cycloartenol, 24-methylencycloartanol, or cyclolandenol (1 parts, particularly about 5 parts) than in the case of single use of the phytosterol. In particular, cycloartenol has synergistic effect on the serum cholesterol lowering action of phytosterol, while 24-methylenecycloartanol and cyclolandenol have weaker effect than that of cycloartenol.

10 Japanese Patent Application Laid-Open No.

27824/1984 reports that when 1% of cycloartenol or 24-methylenecycloartanol was added to a diet S' containing 0.5% of cholesterol, the respective percentages of TC lowering were 13.7% and 10.2% based on a control with high cholesterol diet (calculated by the present inventors from the data *shown in Table 2 of the above patent Laid-Open Gazette).

However, these three patent applications described none of triglyceride (hereinafter referred to as TG), total phospholipid (hereinafter referred "o to as. PL), HDL-C, atherogenic index [(TC HDL-C)/ (HDL-C), hereinafter referred to as AI: some Japanese medical scientists designate the AI as cholesterol ratio or arteriosclerosis index], and lipid peroxide (hereinafter referred to as LPO), though reporting on the action of TC lowering 5 1 in serum.

From the fact that cycloartenol, 24methylenecyclo artanol, and cyclolaudenol, alone or in combination with a phytosterol, depressed TC in serum-, it is not obvious that these triterpenyl alcohols have also the action of lowering the other items of serum lipids TG, PL, and LPO which are important for treating or diagnosing hyperlipidemic conditions, and that these alcohols have the 10 effect of increasing HDL-C, which is currently considered as particularly significant for treating ,I hyperlipidemia, and in addition the effect of lowering AI. It is impossible to predict such overall pharmacological activity from analogy.

The y-orizanol on the market today in Japan as a therapeutic agent for treating whiplash syndrome (head or cervical damage) is not composed of a single component but mixtures of various phytosteryl and I triterpenyl esters of ferulic acid. An example of the y-organol is composed of campesteryl stigmasteryl 8-sitosteryl cycloartanyl cycloartenyl and 24-methylenecycloartonyl esters of ferulic acid but little cyclobranyl ester of ferulic acid.

Recently the following report on the influence of y-orizanol upon cholesterol metabolism in hyperlipidemic rats was published by F. Kuzuya et al.

I 6 S1 (Geriatric Medicine 18, pp 519 524 (1980)).

According to the report; TC was explicitly depressed in rats fed with a high cholesterol diet containing 0.1, 0.5, and 1% of y-orizanol, as compared with TC in control rats fed with the same diet but containing no y-orizanol, while the degree of the lowering depended on the dosage; the degree of TC lowering was greater than that of PL and comparable to that of HDL-C depression; y-orizanol showed no 10 activity an AI, but a tendency to increasing TG and the distinct action lowering LPO.

According to K. Mitani et al. [Domyaku Koka, o 11, No. 2, June, pp 411 416 (1983)], the serum TC values in rats fed with a high chloresterol diet with 0.5, 1,2. and 2.0% of Y-orizanol were lower by 8.1, 23.4, and 30.9%, respectively, than control rats fed with the same diet but containing no Y-orizanol, while no significant depression was observed in the serum TG and PL values.

According to the study of the influence on hyperlipidemia of hypothalamic obesity rats, by Inoue et al. [Domyaku Xoka, 11, No. 2, June, pp.

417 428 (1983)] y-orizanol exhibited the action of lowering TC but not TG in blood and no effect on PL and HDL-C in blood.

As regards organic acids, R.D. Sharma ['Atherosclerosis, 37, pp. 463 468 (1980)] 7 1 describes; that in rats fed with a high cholesterol diet containing 0.2% of an organic acid, TC level was lowered significantly by 10.8% when the acid was ferulic acid, and by 9.4% when the acid was pcoumaric acid, based on TC level in control rats fed with the same diet but containing none of such organic acids; that the degree of TG level lowering was 18.7% with ferulic acid and 19.8% with pcoumaric acid, but these values were not significant; that the PL level lowering was scarcely observed with both the acids; and that no decrease of TC, TG, or PL level was shown with vanillic acid, caffeic acid, or cinnamic acid.

Although an organic acid was not used 15 singly, the following reports on the anti-hyperlipidemic effect of a-methylcinnamic acid derivatives r.t was presented. K. Takashima et al. [Biochemical Pharmacology, 27, 2631 (1978)] describe the antihyperlipidemic effect of a-mono-p-myristyloxy-a'l 20 methylcinnamoyl glycerol. T. Watanabe et al.

4 4 [Journal of Medicinal Chemistry, 23, 50 (1980)] describe in detail synthetic methods of p-alkoxycinnamic acids, p-alkoxy-a-methylcinnamic acids wherein the alkyl moiety in the alkoxy substrituent 25 is 2-propenyl, C 8

C

1 8 alkyl, or phenyl; and m- myristyloxycinnamic acids; m-methoxy-p-alkoxya-methylcinnamic acids wherein the alkyl moiety of

L

8 1 the alkoxy substituent is C 12 or C 1 4 alkyl; p-alkoxycinnamates, and p-alkoxyl-a-methylcinnamates wherein the alkyl moiety of the alkoxy substituent is 2propenyl, methyl, butyl, or C 8

C

18 alkyl and the alcohlic residue of the ester is chloroethyl, metharyloxyethyl, monoglyceride residue, diglyceride residue, etc; and anti-hyperlipidemic activities of these compounds. T. Watanabe et al described also a process for producing p-alkoxy-a-methylcinnamic acids wherein the alkyl moiety of the alkoxy is C-C16 alkyl) (Japanese Patent Publication No.

45582/1976). T. Ota et al. (Japanese Patent Application Laid-Open No. 80370/1982) describes oo0 a° -methyl-p-pyridyloxycinnamic and a-methyl-p- 15 pyridylalkyloxycinnamic acids and (C 1

-C

3 alkyl) a °o esters thereof, processes for producing these compounds, and anti-hyperlipidemic compositions containing these compounds.

Recently, Grill, H. et al. [Japanese Patent S o 20 Application Laid-Open No. 25953/1985); DE, App. No.

a 9 °O 3326164.4 (1983, July, 20)] desribe p-aloxybenzoic acid derivatives such as N-carboxymethyl-4-(2hydroxy-4-phenylbutoxy) benzamide and tert-butylphenyl)-2-oxobutoxy] benzoic acid, 25 processes for producing these derivatives, and anti-hyperlipidemic compositions containing these derivatives.

1 9 1 Also, in the past, attempts have been made to lower the levels of cholesterol, phospholipids, and triglycerides in the blood by the oral feeding of various substances which have been generally -referred to in the art as hypalipidemic agents or hypocholesteremic adjuvants. Several synthetic hypolipidemic agents are now available, namely, clofibrate, D-thyroxine, cholestyramine, and various nicotinic acid-derivatives.

The development of agents capable of reducing elevated blood lipids and of favorably altering blood-lipoprotein patterns is considered by medical authorities to be extremely important for the t a I treatment and prevention of atherosclerosis.

15 The present inventors tested the known compounds cycloartenol, 24-methylenecycloartanol, t°oo* and cyclobranol to ascertain the anti-hyperlipidemic effect thereof. The tests were carried out according to method A (male Wistar strain rats weighing initially 20 100+lg were fed for 2 weeks with the diet limited 9, o* to 10 g/day for each animal but with water given ad libitum) and method B (male Wistar strain rats weighing initially 100+lg were fed for 4 weeks with the diet and water given ad libitum). Details of 25 these test methods will be described later. Results of these tests shown in Tables 1 and 2 (method A) and I- r nr~ 1 Tables 1-1 and 2-1 (method B) were as follows: The hypolipidemic effects according to both methods were fundamentally identical. The decrease of TC in serum was observed in the both group given a hyperlipidemic diet containing cycloartenol and given a hyperlipidemic diet containing cyclobranol, at significance levels (P 0.05 according to method A, P 0.01 according to method as compared with that in the control group given only a hyperlipidemic die't. The TC lowering due to 24-methylenecycloartanol was slight and not significant according to method A but significant (p 0.05) according to method As to HDL-C; cycloartenol depressed it at significant. levels (p 0.05 according to 15 method A, p 0.01 according to method 24methylenecycloartanol lowered it slightly with both method so it was not significant. On the contrary, cyclobranol showed a tendency to increasing HDL-C according to both methods though these increases 20 were not significant. Needless to say, HDL-C level a.

is desired to increase significantly, as shown in the foregoing literature.

4i One of the purposes of the invention is to develop a hypolipidemic agent which significantly o0 25 lowers TC and increases HDL-C in serum. As stated 0 8 above, it was confirmed that one of the triterpenyl alcohols, for instance, cycloartanol, cyclobranol, nr i i, I 11 or 2 4-methylenecycloartanol singly depress TC level in serum significantly. However, no increase in HDL-C content was ascertained in the present inventors' test for hypolipidemic effect according to either methods A or B.

As to AI, cycloartenol and cyclobranol showed tendencies to decrease it according to method A, while 24-methylenecycloartanol showed only a slight tendency to increase. According to method B, the three triterpenyl alcohols showed tendencies to decrease AI. As to TG, PL, and LPO, no significant change was shown with these triterpenyl alcohols according to both methods.

'A 0 Comparing these three triterpenyl alcohols, 15 cyclobranol tended to lower TC, AI, TG, PL, and LPO o but to increase HDL-C, and consequently it was a* different in the action from cycloartenol and 24methylenecycloartanol. That is, it has proved that cyclobranol is superior to cycloartenol and 24- 20 methylenecyloartanol in hypolipidemic effect.

Thus, the present inventors have studied S aiming at the production of an hypolipidemic agent which will decrease the TC, PL, and TG, contents in serum while the HDL-C content is increased, 25 furthermore an agnet which lower AI and LPO contents simultaneously. So our studies have been concentrated on the development of a hypolipidemic agent L 12 1 which has distinctly greater effect in at least 2 3 of 6 items noted above than known triterpenyl alcohols and y-orizanol. As a result, we discovered a number of novel triterpenyl esters of organic acids having excellent hypolipidemic activity. Further we found that each of the three known triterpenyl esters of ferulic acid, certain esters of monobasic saturated fatty acids, and cyclobranol have high hypolipidemic activity singly. It is difficult to predict these facts from properties of each of the known triterpenyl alcohols, organic acids, and y-orizanol.

0 4 0 *04' .SUMMARY OF THE INVENTION 15 It is an object of the present invention to provide novel triterpenyl esters of organic acids except triterpenyl esters of ferulic acid (4hydroxy-3-methoxycinnamic acid), and of monobasic and dibasic saturated fatty acids. More particularly, too k 20 the novel triterpenyl esters of organic acids are organic esters derived from the following alcohols: cycloartenol, cyclobranol, 24-methylenecycloartanol, t ros- Ao\ lanosterol,Kagnosterol, cyclosadol (3 -hydroxy-24methylene-9, 19-cyclo-9-lanosta-23-ene) dihydro- 25 agnosterol, cyclolaudenol, cycloartanol, cycloeucalenol, euphol, butyrospermol, tirucallol, A' N euphorbol, and dammerradienol, except esters of 1

S

i 13 1 ferulic acid, and monobasic and dibasic saturated fatty acids.

Of these esters, preferred are those derived from cycloartenol, 24-methylenecycloartanol, and cyclobranol. Favorable organic acids for the esters are; cinnamic acid, benzoic acid, and a-

(C

1

-C

4 alkyl) cinnamic acid wherein one substituent group on the benzene ring is selected from amino, nitro, hydroxyl, C2-C5 acylamino, C -C4 alkoxy, and

C

2

-C

6 alkylcarboxyl groups; cinnamic acid, benzoic acid, and a-(C-C 4 alkyl) cinnamic acid wherein two substituent groups on the benzene ring are selected from these pairs hydroxyl and C -C4 alkoxy, hydroxyl and C2-C6 alkylcarboxyl, C -C 4 alkoxy and C2-C6 alkylcarboxyl, C -C4 alkoxy and nitro, C1-C4 alkoxy and amino, CI-C4 alkoxy and C2-C5 acylamino, two C1-C alkoxy, two C2-C6 alkylcarboxyl, and two hydroxyl groups; nicotinic acid; and unsaturared fatty acids such as linoleic acid, linolenic acid, 20 arachidonic acid, and eicosapentaenoic acid.

It is another object of the present invention to provide processes for the production of the abovementioned esters of organic acids.

It is still another object of the present 25 invention to provide a hypolipridemic agent which significantly depresses TC and increases HDL-C in serum.

14 r It is still further object of the present invention to provide a pharmaceutical'composition for treating hyperlipidemia, comprising a pharmaceutical carrier and an effective amount of cyclobranol or a triterpenyl ester of organic acid other than dibasic saturated fatty acid.

It is still further object of the present invention to provide a method of treating hyperlipidemia comprising administering to a patient in need of such treatment, a therapeutically effective amount of cyclobranol or a triterpenyl ester of organic acid other than dibasic saturated fatty acid.

According to one aspect of the present invention, there is provided a triterpenyl ester derived from triterpenyl alcohol and organic acid other than ferulic acid and monobasic and dibasic saturated fatty acids, wherein the triterpenyl alcohol is selected from the group consisting of cycloartenol, cyclobranol, 24-methylenecycloartanol, lanosterol, lanostenol, agnosterol, cyclosadol, dihydroagnosterol, cyclolaudenol, cycloartanol, cycloeucalenol, euphol, butyrospermol, tirucallol, euphorbol and dammerdienol and the organic acid is nicotinic acid, linoleic acid, or ;n organic acid of the formula 20 Ar(CH=CR) -COOH n wherein R is hydrogen or a C 1

-C

4 -alkyl group, and Ar is aminophenyl, 0* nitrophenyl, hydroxyphenyl, a C 1

-C

4 -alkoxyphenyl, C 1

-C

4 -alkyl-CONHphenyl,

(C

1

-C

5 -alkyl)COO-phenyl, or Ar is a phenyl substituted by C 1

-C

4 -alkoxy and hydroxy, by C 1

-C

5 -alkyl-COO and hydroxy, by C 1

-C

4 alkoxy and (CI-C 5 alkyl)COO, by C 1

-C

4 -alkoxy and nitro, by C 1

-C

4 -alkoxy and amino, or by

C

1

-C

4 -alkyl-CONH and C 1

-C

4 -alkoxy or Ar is di-(C 1

-C

4 -alkoxy)phenyl, di(Cl-C 5 -alkyl-COO)phenyl or dihydroxyphenyl; and n is 0 or 1.

According to another aspect of the present invention, there is provided a process for producing a triterpenyl ester of organic acid ester 313 other than esters of ferulic acid and of monobasic and dibasic saturated fatty acids, which comprises the reaction of a triterpenyl alcohol with an acid halide of the corresponding organic acid.

According to still another aspect of the present invention, there is S provided a pharmaceutical composition for treatment of hyperlipidemia 3.5 comprising o o g0 0 o 06 a *0*o o 0 00 0 0 0 jA. TMR/866c 15 1 a pharmaceutical carrier and an effective amount of a triterpenyl ester of organic acid other than triterpenyl esters of dibasic saturated fatty acids.

According to still another aspect of the present invention, there is provided a phermaceutical composition for treating hyperlipidemia which comprises a pharmaceutical carrier and an effective amount of cyclobranol as an active ingredient.

According to still another aspect of the present invention, there is provided a method of treating hyperlipidemia comprising administrating to a patient in need of such treatment, a therapentically effective amount of cyclobranol, or of a o triterpenyl ester of organic acid other than dibasic 15 saturated fatty acid.

It tt itt I DESCRIPTION OF THE PREFERRED EMBODIMENTS The novel compounds of the present invention are, in general, white crystalline solids having t e S 20 characteristic melting points and specific rotation, and stabje compounds which, as can be seen from later examples of preparation, that those are not hydrolyzed Sat all even heated at 60 70 0 C for 3 hours in a strongly acidic aqueous solution of pH 0.5 25 with stirring.

Structural formulae of preferred three triterpenyl esters of organic acids are shown below.

r 2 i:~ Ia

CH

3

RO

CH

3 'CH 3

CHR

2 H 3C CH3 3 CH 3 Ib RO C CH 3 CR 3 CH 3 H

C

CH Ic

CHH

RO CH 3

CH

3 6H 3 When R is N in the general formulae Ia, Tb, and Ic, formula Ia represents cycloartenol, formula lb 24-methylenecycloartano1, and formula Ic ,tt' 4t 4 ('1 Lui; 17 1 cyclobranol. These three triterpenyl alcohols are publicly known.

In the present invention, R of formulae Ia, Ib, and Ic herein represents a residue of the abovementioned various monobasic organic acids. The following general formulae II and IIIa IIId represent the compounds of the present invention which have, in the molecule, one of these organic acid residues except nicotinic acid, linoleic acid, linolenic acid, arachidonic acid, and eicosanpentaenoic acid residues.

General formula R -triterpenyl alcohol residue R II R II.

6 6 6 In formula II, R 1 denotes a,-unsaturated carbonyl group carbonyl group or a-(C1-C 4 alkyl) a,6-unsaturated carbonyl group

(-CH=CR

3 and R denotes amino acylamino

(-NHCOR

3 nitro (-NO 2 hydroxyl C 1

-C

4 alkoxy or C 2

-C

6 alkylcarboxyl (-OCOR 4 R is

C

1

-C

4 alkyl, i.e. methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl, and R4 is 25 C1-C 5 alkyl, that is, any of the above alkyls denoted 1 5 by R 3 and of the C 5 alkyls, i.e. pentyl iso-pentyl, sec-pentyl, 3-pentyl, and tert-pentyl.

I

18 General formula (II) represents any of the triterpenyl'esters of cinnamic acid, benzoic acid, and x- (C 1 -C 4 alkyl) cinnamic acid which have the subs ti tue: R on the ortho-, meta-, or para- position the benzene ring.

General formula R 1-trtrey alcohol residue

OH

OR 3or OCOR 4 IIa It t tv t i xt 0 t R 1-triterpenyl alcohol residue OR3 OCOR 4

NO

2 NH 2 or NHCOR 3 R1- riterpenyl alcohol residue 2 II lb I IIc alcohol residue (OR 3 2 IIId In general formulae Il~a d, R 1 R 3 and Rare as defined above.

The compound of general formula Ila is a triterpenyl ester of cinamic acid, benzoic acid, or c-(C 1 -C 4 alkyl)cinnamic acid each having, on the I~I- 'a 19 1 benzene ring, two different substituents OH and OR 3 groups, or OH and OCOR 4 groups. The compound of general formula IIIb is the same ester but the acid residue of which has on the benzene ring, two different substituents OR 3 and OCOR groups, OR 3 and

NO

2 groups, OR 3 and NH 2 groupos, or OR 3 and NHCOR 3 groups.

Referring to the compounds of formula IIIa or IIIb, detailed description is given below.

The compound of formula IIIa, when an OH group is attached to the benzene ring at theoposition (2-position), is an ester having an OR 3 or OCOR 4 group at the 5- or 6-position.

When an OH group is attached to the benzene ring at 15 the m-position (3-position), the compound is an i« a ester having an OR 3 or OCOR 4 group at the or .6-position. When an OH group is attached to the benzene ring at the p-position (4-position), the compound is an ester having an OR 3 or OCOR 4 group 20 at the 2- or 3-position (hereinafter these compounds I* are referred to as compounds of formula IIIa).

P. i Such bond-structures of the compounds of formula IIIa are shown by the following general formulae IIIa IIIal 0 1 10 L; 20 1General formula R 1 triterpenyl alcohol residue ~OHMa OR 3 or-OCOR 4

R

1 -triterpenyl alcohol residue ~OH IIIa2

O

3 or -OCOR 4 Rj-triterpenyl alcohol residue -OH 111a3 RO0 o-CO or4 CR 4 ltriterpenyl alcohol residue

R

3 0OH 4 t 1 tiepnl loo eiu -21 R 1-triterpenyl alcohol residue OH III a6 OR 3 or -OCOR 4 R 1-triterpenyl alcohol residue 7 III a7 RO3 OH o -OCR 4 R -triterpenyl alcohol residue RO3 0 Ija8 o OCOR4

H

R triterpenyl alcohol residue aOR 3or -OCOR 4IIIa9 3

OH

-triterpenyl alcohol residue f t IIlalO "OR 3 or -OCOR 4

OH

22 1 In formulae IIIal IIIalO, R 1

R

3 and R are as defined above.

The compound of general formula IIIb is a tri-erpenyl ester of cinnamic acid, benzoic acid, or a-(C 1

-C

4 alkyl) cinnamic acid each having, on the benzene ring, two different groups, i.e. an OR 3 group, in place of the OH group of the compounds of formula IIIa, and any of OCOR 4

NO

2

NH

2 and

NHCOR

3 groups. Consequently, the compound of formula IIIbl shown below is a compound resulting from the substitution of an OR 3 group for the OH group of the compound of formula IIIal and the substitution of an OCOR 4

NO

2

NH

2 or NHCOR 3 group e for the OR 3 or OCOR 4 group of the compound of formula 15 IIIal.

a* The compounds of the following formulae a IIIb2 IIIblO have similar bond-structures. In these formulae, X denotes OCOR 4

NO

2

NH

2 or NHCOR 3 group.

4.I 4 tt SR-triterpenyl alcohol residue SOR IIIbl 9R 3 I X2 V r 23 R 1 triterpenyl alcohol residue II1b2 R 1 -triterpenyl alcohol residue

OR

3 II1b3 R -triterpenyl alcohol residue x OR3 II1b4 &~tt 9 0# fs

I

9 R 1-triterpenyl alcohol residue R 1-triterpenyl alcohol residue IIub6 I It

I

24 R 1 triterpenyl alcohol residue IIlb 7 OR 3 R triterpenyl alcohol residue x OR 3 R triterpenyl alcohol residue I x II1b8 ci of 0~ 4 Ck 0*"0 0* 0 0 a 0 a j a 00 O OR 3 alcohol residue IIlb 9 IlIblO In formulae IIIbl IlIblO, R 1 and R 3are as defined above.

The compound of general formula IlIc and the compound of general formula IIId are triterpenyl esters of cinnamic acid, benzoic acid, or a-(C 1 -C 4 alkyl) cinnamic acid having, on the benzene ring, 252 1 two OH groups and two OR 3 groups, respectively.

That is, the compound of formula IIIc has two OH groups at the 2- and 3-positions, 2- and 4-positions, 2- and 5-positions, 2- and 6-positions, 3- and 4positions, or 3- and 5-positions, that is, the following six bond-structures are present for the compounds of formula IIIc.

SRl-triterpenyl alcohol residue

OH

OH [IIIcl

OH

0 0 15 R1-triterpenyl alcohol residue o OH IIIc2

OH

R triterpenyl alcohol residue

OH

IIIc3 1 L°o o HO iHO 0 1 t t t

I

r 26 R -triterpenyl alcohol residue I HO IIIc4 R,-triterpenyl alcohol residue

OH

OHtriterpenyl alcohol residue

R

1 -triterpenyl alcohol residue @oro

C

C

t a at, it a i c a a a at aD I iaa a a i a( a a Ca ar

I

HO

IIIc6

A-"

-7 s- The compound of formula IIId has two OR 3 groups in place of the two OH groups of the compound of formula IIIc, thus including six compounds of 20 formulae IIIdl IIId6 similarly to formulae IIIcl IIIc6.

In the next place, the process for producing the compound of the invention is described.

The aforementioned Y-orizanol is a suitable raw material for cycloartenol, 24-methylenecycloartanol, and cyclobranol. The y-orizanol, as stated before, is not a single compound but a mixture of BI _-i 2~i12::7al

II

27 C r~o i D D 1 various steryl and triterpenyl esters of ferulic acid. The mixture, for example, is composed of campesteryl stigmasteryl -sitosteryl cycloartanyl cycloartenyl and of 2 4-methylenecycloartanyl esters of ferulic acid. With reference to the method of Endo et al.

(Yukagaku, 18, pp. 63 67 (1969)), y-orizanol was recrystallized repeatedly by using acetone-methanol (methanol content 2 acetone, and ethyl acetate to give cycloartenyl ester of ferulic acid, which was then saponified to separate cycloartenol, m.p. 101 102 0 C, specific r ation [a]21.5 49.7° (c 1.01, CHCR 3 This cycloartenol, on gas chromatography, gave a single peak.

Method for isolating 24-methylenecycloartanol: Accordingto the above method of Endo et al., crystals obtained from the mother liquor after separating cycloartenol from y-orizanol were 20 acetylated with phridine-acetic anhydride, the acetylated product was recrystallized repeatedly by using chloroform-ethyl acetate-ethanol 322) and then deacetylated, and the product was recrystallized from acetone-methanol to give 24-methylenecycloartanyl ester of ferulic, acid, which was then saponified to isolate 24-methylenecycloartanol, 24 m.p. 123 124 0 C, specific rotation D 48.10 v 28 1 (c 1.00, CHC 3 This substance, on gas chromatography, gave a single peak.

Method for isolating cyclobranol: y-Orizanol (1.1 Kg, cyclobranol content 0%) was dissolved in acetone (8 After dissolution of iodine (40 g) therein, the mixture was heated for 1.5 hours under reflux. The mixture was then allowed to cool, a 10% aqueous solution (500 ml) of sodium thiosulfate was added, the mixture was stirred for 30 minutes, and further water (550 ml) was added. The formed crystals were filtered, washed wit 2% aqueous solution (700 ml) of sodium thiosulfate and then with water (4 and soo oe.o" dried to give a y-orizanol (1 Kg), which was found oa 9o by gas chromatography to contain about 23% cyclo- ***ooo Sbranol. This crystalline product (1 Kg) was suspended in a 4% ethanolic KOH solution, and the o00 ooB 0 suspension was heated for 3 hours under reflux.

After cooling the resulting mixture, the precipitated S.o 20 potassium salt of y-orizanol was filtered, and suspended in methanol (8 and the suspension was refluxed for 2 hours. After cooling the resulting S mixture, the precipitated yellow crystals were filtered and dried to give the potassium salt of 25 a y-orizanol (260 This crystalline product was Streated similarly with 3% and 2% ethanolic KOH solutions to give yellow crystals (130 This Lx-~ 29 0~40 i 0 I C t S Cii C t 1 product was found to contain 88% cyclobranol.

Further this yellow crystalline product (130 g) was saponified in a 2 N ethanolic KOH solution (2.6 and the residue was extracted with chloroform (1.2 The extract was dried and evaporated under reduced pressure to give crude cyclobranol (80 g, 88% purity), which was then recrystallized 3 times from acetone (1.6 thus giving crystalline cyclobranol (28 m.p. 165 25 166 specific rotation D 47.0° (C 1.00,

CHCZ

3 This cyclobranol, on gas chromatography, gave a single peak.

The triterpenyl esters of organic acids according to the present invention can be readily obtained by known methods of esterification from the above defined alcohols and organic acids. That is, the ester can be prepared by the esterification of the organic acid and the triterpenyl alcohol through dehydration in the presence of a catalyst such as sulfuric acid, p-toluenesulfonic acid, or boron trifluoride (BF 3 the reaction of the anhydride of the organic acid with the triterpenyl alcohol in the presence of a catalyst such as sulfuric acid or zinc chloride, or the reaction of a halide of 25 the organic acid (the corresponding acyl halide; hereinafter referred to as organic acid halide),

-A

3 a

C

C aC a.

a.' Swith the triterpenyl alcohol. Of these methods, the most favorable is the method of reacting the organic acid halide with the triterpenyl alcohol.

That is, when the starting organic acid is a monobasic acid such as nicotinic acid, linoleic acid, linolenic acid, arachidonic acid, eicosanpentaenoic acid, C 6

C

14 saturated fatty acids, or an organic acid corresponding to an ester of formula II, IIIB, or IIId, i.e. a mono- or disubstituted cinnamic, benzoic, or a-(C 1

-C

4 alkyl) cinnamic acid having; one substituent selected from NO 2

OR

3

OCOR

4 and NHCOR 3 groups (formula II); two different substituents OR 3 group and any of OCOR NO 2

NH

2 and NHCOR 3 groups (formula IIIb); or two OR 3 groups (formula IIId); on the benzene ring; the intended triterpenyl esters of organic acid can be obtained with ease and in a high yield by converting the COOH group of the starting organic acid with a halogenating reagent into the CO-halogen group, followed by esterifying the resulting acid halide with the triterpenyl alcohol in the presence of a dehydrohalogenating agent in a solvent at a temperature of 10 to 100 0 C. Favorable halogenating reagents for this purpose are thionyl chloride, sulfuryl chloride, phosphorus pentachloride, phosphorus oxychloride, benzoyl chloride, phthaloyl chloride, hydrogen chloride, and hydrogen bromide.

31 1 Suitable dehydrohalogenating agents for use in the esterification are pyridine, quinoline, trimethylamine, triethylamine, tripropylamine, tributylamine, magnesium, and dimethylaniline.

When the starting organic.acid is one corresponding to an ester of formula II, IIIa, or IIIc a mono- or di-substituted cinnamic, benzoic, or a-(C1-C4 alkyl) cinnamic acid having one substituent OH or NH 2 group (formula II), two different substituents OH and OR 3 groups or OH and

OCOR

4 groups (formula IIIa), or two OH groups (formula IIIc), on the benzene ring], the acylated derivative of triterpenyl esters of organic acids can be obtained with ease and in a high yield by acylating the OH or NH 2 group of the organic acid Sin advance, followed by halogenation and esterification of the resulting acid as stated above.

Then each ester of formula II, IIIa, or IIIc having OH or NH 2 group on the benzene ring can be prepared 20 by deacylating the above acylated derivative, that is, by heating it in a concentrated aqueous solution of ammonia, caustic alkali (NaOH or KOH), or inorganic acid (HC1, H 2 S0 4 or H 3

PO

4 The acylation can be readily accomplished 25 by using an acylating agent such as an acid anhydride 4* or acid halide of such a lower fatty acid as acetic acid, propionic acid, butyric acid or caproic acid.

i 32 1 Further an ester of formula II or IIIb having one substituent NH 2 group or two different substituents NH 2 and OR 3 groups or NH 2 and OCOR 4 groups on the benzene ring can be prepared by reducing the corresponding triterpenyl esters of substituted cinnamic, benzoic, or a-(C 1

-C

4 alkyl) cinnamic acid having one NO 2 group, NO 2 and OR 3 groups, or NO 2 and OCOR 4 groups, on the benzene ring of the acid portion, with iron (or zinc) and acid (HC1, H 2

SO

4 or acetic acid), or with tin (or tin chloride) and conc. H2SO4 to convert the NO 2 group selectively into NH 2 group. This reduction method using metal and acid is best suited since the unsaturated group present in the triterpenyl e alcohol portion is not reduced.

p G The acylation of the above amino derivatives in the usual manner gives readily the corresponding esters of the invention having one NHCOR 3 group or two groups of NHCOR 3 and either OR 3 or OCOR 4 on the 20 benzene ring of the acid portion.

;I Pharmacological action: In the next place, detailed description is given on the results of pharmacological tests of compounds according to the present invention for toxicity and anti-hyperlipidemic activity.

SAcute toxicity test: Acute toxicity tests were conducted using

]I'

-M r I.1 33 male ddy strain mice (30 2 g) and 5 male Wistar strain rats (100 2 g) for each compound by oral administration.

Examples of the compounds according to the present invention subjected to the acute toxicity tests were as follows: Example 29, Cycloartenyl ester of 4-hydroxy- 3-methoxybenzoic acid, Example 37, Cyclobranyl ester of 4-hydroxy- 3-methoxybenzoic acid, Example 86, 24-Methylenecycloartanyl ester of 4-hydroxy-3-methoxybenzoic acid, Example 60, Cycloartenyl ester of 3-ethoxy- 4-hydroxybenzoic acid, Example 62, Cyclobranyl ester of 3-ethoxy- 4-hydroxybenzoic acid, Example 74, Cycloartenyl ester of 4-hydroxy- 3-propoxycinnamic acid, Example 71, Cyclobranyl ester of 3-ethoxy- 20 4-hydroxycinnamic acid, Example 27, Cycloartenyl ester of 3,4t.a dihydroxybenzoic acid, Example 39, Cyclobranyl ester of 3,4dihydroxybenzoic acid, 25 Example 83, 24-Methylenecycloartanyl ester of p-acetoxycinnamic acid, r 34 I Example 2, Cycloartenyl ester of 3,4dihydroxycinnamic acid, Example 32, Cyclobranyl ester of 3,4dihydroxycinnamic acid, Example 82, 24-Methylenecycloartanyl ester of 3,4-dihydroxycinnamic acid, Example 8, Cycloartenyl ester of o-hydroxybenzoic acid, Example 41, Cyclobranyl ester of o-hydroxybenzoic acid, Example 72, 24-Methylenecycloartanyl ester of 4-acetoxy- 3.-ethoxycinnamic acid, Example 10, Cycloartenyl ester of phydroxybenzoic acid, Example 58, Cyclobrany. ester of phydroxybenzoic acid, Example 13, Cycloartenyl ester of o-methoxybenzoic acid, Example 14, Cycloartenyl ester of p- 20 methoxybenzoic acid, Example 24, Cycloartenyl ester of nicotinic acid, Example 50, Cyclobranyl ester of nicotinic f ti Ct '.4 Ce 4 4 444 '9 14 t 4 4$ acid, Ex~ample 20, Cycloartenyl ester of pacetamidobenzoic acid, 1 Example 19, Cycloartenyl ester of p-aminobenzoic acid, Example 43, Cyclobranyl ester of pamiriobenzoic acid, Example 100, 24-Methylenecyloartanyl ester of m-aminobenzoic acid, Example 25, Cycloartenyl ester of linoleic acid, Example 51, Cyclobranyl ester of linoleic acid, Example 93, 24-Methylenecyloartanyl ester of linoleic acid, Example 12, Cycloartenyl ester of mhydroxybenzoic acid, 4 1 C Example 54, Cyclobranyl ester of mhydroxybenzoic acid, Example 16, Cycloartenyl ester of onitrobenzoic acid, Example 47, Cyclobranyl ester of oazninobenzoic acid, Example 23, Cycloartenyl ester of maminobenzoic acid, Example 49, Cyclobrany. ester of maminobenzoic acid, Example 100-1, Cycloartenyl ester of 4hydroxy-3-methoxycinnamic acid (another name: cycloartenyl ester of ferulic acid), -36 1 Example 100-2, Cyclobranyl ester of 4hydroxy-3-methoxycinnamic acid, Example 100-3, 24-Methyleriecycloartanyl ester of 4-hydroxy-3-methoxycinnamic acid, Example 101, Cycloartenyl ester of pnitrocinnamic acid, Example 102, Cycloartenyl ester of paminocinnamic acid, Example 104, Cyclobranyl ester of paminocirinamic acid, Example 108, Cycloartenyl ester of maminocinnamic acid, Example 112, 24-Methylenecyloartanyl ester of m-aminocinnamic acid, Example 114, Cycloartenyl ester of 4hydroxy-3-methoxy-a-methylcinnamic acid, Example 116, Cyclobranyl ester of 4-hydroxy- 3-methoxy-oa-methylcinnamic acid, Example 118, 24-Methylenecycloartanyl ester of 4-hydroxy-3-methoxy-x-methylciinamic acid, 4 Example 120, Cycloartenyl ester of 4-hydroxy- 3-methoxy-ct-ethylcinnamic acid, Example 140, Cyclobranyl ester of 3-ethoxy- 4-hydroxy-ca-methylcinnamic acid, Example 130, Cycloartenyl ester of 4-hydroxyct-ethylcinnamic acid, 37 1 Example 146, Cycloartenyl ester of 4-hydroxy- 3-propoxy-a-methylcinnamic acid, Example 167, Cycloartenyl ester of 4-amino- 3-methoxybenzoic acid, Example 173, Cyclobranyl ester of 2-methoxybenzoic acid, Example 189, Cycloartenyl ester of 4-amino- 3-methoxy-ca-methylcinnamic acid, Example 177, Cycloartenyl ester of 4-amino- 3-methoxycinnamic acid, Example 205, Cycloartenyl ester of p-aminoa-methylcinnamic acid, Example 183, Cycloartenyl ester of o 2-ethoxycinnamic acid, Example 212, Cyclobranyl ester of in-aminoc-methylcinnamic acid, Example 191, 24-Methylenecycloartanyl ester of 4-amino-3-methoxy-ca-methylcinnamic acid, Example 197, 24-Methylenecycloartanyl ester air 1 20 of 5-amino-2-propoxy-a-methylcinnamic acid, to Example 171, Cycloartenyl ester of 2-methoxybenzoic acid, Example 170, Cycloartenyl ester of 2-methoxy- 5-nitrobenzoic acid, 25 Example 178, Cyclobranyl ester of 4-amino- 3-methoxycinnamic acid, 38 1 Example 195, Cycloartenyl ester of 2-propoxy-a-methylcinnamic acid, Example 213, 24-Methylenecycloartanyl ester of m-amino-a-methylcinnamic acid, Example 113, Cycloartenyl ester of 3-methoxy- 4-propionyloxy-a-methylcinnamic acid, Example 117, 24-Methylenecycloartanyl ester of 3-methoxy-4-propionyloxy-a-methylcinnamic acid.

The above 66 compounds of esters, and as 2yc- ,e+yleecycoir- n o\ control drugs, cycloartenol, cyclobranol, and y-orizanol were given forcibly to the above-mentioned mice and rats by means of a throat explorer rod in doses of 0.1 to 5 g/Kg for mice and 2 to 6 g/Kg for rats. During the test, the temperature of the *n 15 animal room was kept at 22 to 23 OC. After administration, the animals were observed for 2 weeks. With these doses none of the animals died. During the observation, no symptom of toxicosis appeared and no difference in behavior as well as in body weight was found between the animals given the test and the normal animals not given the compound. In the inspection conducted after 2-week observation, no macroscopic lesion was found in any part of main organs. Thus the compound of the invention has very 25 low toxicity, so that the LD 50 value could not determined.

I~ 39 Pharmacological test method A for anti-hyper lipidemic activity (the term "method A" is used in the present specification) Male Wistar strain rats (100 Ig, each rats were formed into a group) were used as test animals. Diet for the control group was prepared by thorough mixing of 20% casein, 62.5% glucose, hydrogenated coconut oil, 2% agar powder, 4% vitamin-containing salt mixture, 1% cholesterol, and 0.5% cholic acid (this composition is described by Fukushima et al, in "Yakugaku Zassi", 89, No. 6, pp. 857 862 (1962)). Diet for the test groups was prepared by good mixing of 1% each of cycloartenol, 24-methylenecycloartanol, and cyclobranol with the control group's diet. Each rat was kept in a cage S* at a constant temperature of 23 1C and a constant SR.H. of 55 5% for 2 weeks, during which 10 g/day of prescribed was given. Finally the rats were fast,- from food except water for 16 hours (from 4 p.m. of the 14th test day to 8 a.m. of the test day), and under anesthesia with pentbarbital sodium (tradename: Nembutal), blood was taken from each rat through the descending abdominal arota.

i Then, TC, HDL-C, TG, PL, and LPO in the serum were 4*A 25 measured in the manner that will be described below.

Pharmacological test method B for antihyperlipidemic acitivity (the term "method B"

~I

40 O 0 4099*4 Ir I I 0 ;r I r Ir

I~P

(FI

1 was used in the present specification) Male Wistar strain rats (100 1 g, each 8 rats were formed into a group except that the control group fed with a hyperlipidemic diet was formed of 16 rats) were used as test animals. A powdery diet (CE-2, supplied by Clea Japan, Inc.) was used as ordinary diet. The hyperlipidemic diet was prepared by fortifying the ordinary diet with cholesterol and cholic acid Each test compound for administration was admixed with the hyperlipidemic diet. Each two rats were kept in a cage and given the prescribed diet and water ad libitum. Thus the rats were fed for 4 weeks at a constant temperature of 23 1 0 C and a constant 15 R.H. of 55 Finally, the rats were fasted from food except water for 16 hours (from 4 p.m. of the 28th test day to 8 a.m. of the 29th test day), and under anesthesia with pentbarbital sodium (tradename: Nembutal), blood was taken from each rat through the descending abdominal arota. Then, TC, HDL-C, TG, PL, and LPO in the serum were measured in the following manner.

Method for determination of serum TC A TC kit-K (supplied by Nippon Shoji Kaisha, Ltd.) was used. The principle of this determination is as follows: The ester of cholesterol in the serum is hydrolyzed with cholesterol-ester hydrolase into P 41 1 free cholesterol and fatty acids. All the free cholesterol is oxidized with cholesterol oxydase to form 4 -cholestenone and hydrogen peroxide. Phenol and 4-aminoantipyrin are oxidatively condensed together by the formed hydrogen peroxide and peroxydase. The produced red quinone coloring matter is measured by colorimetry for absorbance at 500 nm using a spectrophotometer, thereby determining the TC: Preparation of color-developing liquid: Color-developing reagent: One bial (components: cholesterol esterase 25 ,000u,cholesterol oxydase 25 u peroxydase 3,554 4-aminoantipyrin 20 mg) 15 Buffer solution: 100 ml of the solution S° contains phenol (33.3 mg), potassium dihydrogenphosphate (489.9 mg), and anhydrous disodium hydrogenphosphate (908.5 mg) in purified water.

Standard solution: 100 ml of the solution 4 .0 20 contains cholesterol (300 mg).

A solution of one bial of the above colordeveloping reagent in 160 ml of the buffer solution is referred to as color-developing liquid.

The color-developing liquid (3.0 ml) is well 25 mixed with the sample serum (0.02 ml). The mixture once heated at 37 0 C for 15 minutes, is measured for

IF

I i: i *4*S 1 4 I S

S

42 1 absorbance at 500 nm with a spectrophotometer.

The found absorbance is denoted by EA. On the other hand, the color-developing liquid (3.0 ml) is well mixed with the standard solution. The mixture is treated and measured for absorbance at 500 nm in the same manner as the above. This found absorbance is denoted by ES. Both EA and ES are determined with reference to the value of the blank test conducted using the color-developing liquid (3.0 ml) alone.

EA

TC value (mg/d) x 300 mg/d

ES

Method for determination of serum HDL-C An HDL-C kit-N (supplied by Nippon Shoji Kaisha, Ltd.) was used. Ultra-high density lipoprotein (VLDL) and low density lipoprotein (LDL) in the serum is precipitated by the action of heparin. The precipitate is separated by centrifugation. High density lipoprotein (HDL) is dissolved in the separated supernatant. Esters of cholesterol in this fraction is hydrolyzed with cholesterol-ester hydrolase into free cholesterol and fatty acids.

All the free cholesterol is oxidized with cholesterol oxydase to form A -cholestenone and hydrogen peroxide. Phenol and 4-aminoantipyrine are oxidatively 25 condensed together by the formed hydrogen peroxide and peroxydase. The produced red quinone coloring matter is measured by colorimetry for absorbance

S.

1 11I 0S

I

11 5

S

i: 1 1- 43 1 at 500 nm using a spectrophotometer, thereby determining the HDL-C.

Method for determination of serum PL: A PL kit-K (supplied by Nippon Shoji Kaisha, Ltd.) was used for the determination. Lecithin, sphingomyelin, and lysolecithin are decomposed by phospholipase D into choline and phosphatidic acid, N-acylsphingosyl phosphate, or lysophotphatidic acid, respectively. The resulting choline is quantitatively decomposed by choline oxydase into hydrogen peroxide and betaine. With this hydrogen peroxide, and aid of peroxydase, phenol and 4o aminoantipyrin are condensed into red quinone pigment, then the absorbance at 500 nm of which is 15 measured with a spectrophotometer, thereby determining the PL.

Method for determination of serum TG: Serum TG level was determined by means of a triglycerlde test kit (supplied by Wako Pure Chemical Industries, Ltd.) in which acetylacetone is used for a reagent, in the following manner: Serum proteins are precipitated, when isopropyl alcohol and the sample serum are mixed. Thereby t serum lipids and sacchalides are extracted into the 25 isopropyl alcohol layer. An adsorbent is added to the isopropyl alcohol solution to adsorb colorationinterferring materials. After centrifugation of IC" i i~i 1 i 44 .t 1 9L i i 4i 4 the mixture, potassium hydroxide is added to a portion of the supernatant, thereby saponifying the triglyceride to liberate glycerol. Then the pH of the mixture is adjusted to 6 by adding a buffer solution, and a sodium metaperiodate solution is added to oxidize the glycerol into formic acid (1 mole from mole of gylcerol) and formaldehyde (2 moles from mole of glycerol). The resulting form aldehyde is allowed to react with acetylacetone and with the ammonia in the buffer solution, forming a cyclic compound 3,5-diacetyl-l,4-dihydrobutidine.

This yellow pigment is determined by measuring the absorbance at 410 nm with a spectrophotometer, thus determining the TG content.

Method for determination of serum LPO: A lipoperoxide test kit (supplied by Wako Pure Chemical Industries, Ltd.) according to the Yagi's thiobarbituric acid method Yagi, Biochem.

Med. 15, p 212 (1976), Vitamin 49, p 403 (1975)] was used for this determination. Physiological saline (1.0 ml) is added to the sample serum (0.05 ml) and the mixture is stirred gently. After centrifugation (3,000 10 minutes) of the mixture, 1/12 N-H 2

SO

4 (4.0 ml) is added and well mixed with the supernatant (0.5 ml). A 10% aqueous phosphotungstic acid solution (0.5 ml) is added 45 a 4L

I

I W %4 1 thereto and the mixture is stirred well, allowed to stand for 5 minutes, and centrifuged at 3,000 r.p.m.

for 10 minutes. The resulting precipitate is suspended thoroughly in a mixture of 1/12 NH2SO4 (2.0 ml) and a 10% aqueous phosphotungstic acid solution by means of a mixer. The suspension is then centrifuged at 3000 r.p.m. for 10 minutes, then the obtained precipitate is suspended in distilled water (4.0 ml) by means of a mixer. Then a TBA reagent (1.0 ml; 50% acetic acid solution containing thiobarbituric acid) is well mixed with the suspension.

The mixture is charged in a centrifuge tube, and heated in a boiling water bath for 60 minutes with a glass ball placed on the top of the tube. After 15 5-minute cooling of the tube in a running water, butanol (5.0 ml) is added to the mixture, and well mixed for 20 seconds by means of a mixer with the tube being stoppered, thereby extracting the reaction product in the butanol. The mixture is centrifuged at 3000 r.p.m. for 10 minutes. The butanol layer is measured for fluorescence. After adjustment of the zero point by a blank test, the fluorescence intensity of 0.1 ml of a standard solution (1,1,3,3-tetraethoxypropane 5 n mole/ml) and that of the sample are measured at 553 nm with an excitation wavelength of 515 nm. That is, in this method, the product of the reaction of LPO I 2- F- li P. I I I I-L- i ~l II I- ll 46 044* 0 4 9 9 44 4 4 (0 4 144 4( 4 1 with thiobarbituric acid is identical with that of malondialdehyde with thiobarbituric acid. Accordingly, the LPO concentration is determined as the amount of malondialdehyde in 1 ml of the serum. The standard solution is a 5n mole/ml aqueous solution of 1,1,3,3tetraethoxypropane, which is converted quantitatively into malondialdehyde. Since 0.1 ml of the standard solution is used in this method, the quantity of l,1,3,3-tetraethoxypropane used is 0.5 n mole.

Consequently the LPO content is calculated according to the following equation: LPO content (n mole/ml serum) f 1.0 1.05 f 0.5 x f x x f x 21 F 0.05 0.5 F Results of phamacological tests for hypolipidemic activity: The hypolipidemic action of typical compounds among the present invention on serum lipids and on serum lipid hydroperoxide are described below. The compound selected here are the same as selected in 20 2the description of the acute toxicity. The tests were carried according to the above methods wherein rats were fed with high cholesterol diet.

Results of the tests for hypolipidemic activities of cycloartenol, cyclobranol, and 24- 25 methylenecycloartanol, which are used as control drugs, are shown in Tables 1 and 2 (according to method A) and Tables 1-1 and 2-1 (according to method B).

47 1 Effects of these compounds were described already.

Hypolipidemic effects of compounds of the present invention, tested according to method A are shown in Tables 3 to 14, 14-1, and 14-2. The effects of compounds according to the present invention and those of cycloartenol, cyclobranol, 24-methylenecycloartanol, and y-orizanol as control drugs, with method B are shown in Tables 3-1 to 8-1. Tables 3-1 to 8-1 indicate that TC, PL, and LPO of the normal diet feeding group (denoted by N) were depressed at high significance levels (p 0.001, marked with without exception while the HDL-C was raised at high o° significance levels (p 0.001), in contrast with o* those of the control group fed with the hyperlipidemic 15 diet (denoted by On the TG in N tendency to o depress was shown in C, but this difference between values of TG in N and C was not significant.

Hypolipidemic effects of improving serum lipids' components were clearly observed in the groups 20 given the hyperlipidemic diet containing each of compounds according to the present invention, or each of the control drugs, as compared with those in the group given the hyperlipidemic diet only. In particular, compounds according to the present 25 invention brought about distinctly better effects on two or more serum lipids components of TC, HDL-C, PL, and LPO than did the control compounds.

48 1 According to method A, the TC level were changed by compounds of the present invention as follows: The compound of Example 49 depressed the TC levels at a high significance level (p 0.001).

The compounds of Examples 37, 62, 71, 27, 39, 8, 41, 58, 13, 14, 19, 43, 100, 25, 51, 93, 100-1, 100-2, 101, 105, and 109 depressed the TC levels at medium significance levels. The compounds of Examples 29, 86, 60, 74, 83, 32, 72, 24, 50, 20, 12, 16, and 23 depressed the TC levels at low significance levels (p 0.05). The compounds of Examples 2, 82, and 47 didn't depress the TC significantly, but tended to j depress apparently.

According to method B, the TC levels were |t 15 depressed by the control drugs, i.e. the three triterpenyl alcohols and y-orizanol, at medium significance levels (p 0.01), as compared with the TC levels in the control group given the hyperlipidemic diet only. In contrast, the compounds of Examples 114, 116, 118, 189, 205, 212, 191, 197, 171, 178, 195, and 23.3 depressed the TC levels at high significance S levels (p 0.001). The compounds of Examples 120, 140, 130, 146, 167, 173, 177, 183, 170, 113 and 117 depressed the TC levels at medium significance S 25 levels (p 0.01).

The HLD-C contents, according to method A, were affected by compounds of the invention as follows: i 49 1 The compound of Example 8 raised the HLD-C contents at a high significance level (p 0.001), the compounds of Examples 37, 62, and 39 at medium significance levels (p 0.01), and the compounds of Examples 71, 27, 83, 41, 51, 100-1, 105, and 109 at low significance levels (p 0.05). The compounds of Examples 86, 74, 10, 12, 47, and 49 scarcely varied or slight depressed the HLD-C contents while the compounds of other Examples showed tendencies to raise them but insignificantly.

According to method B, effects of test compounds on the HLD-C were as follows: The control drug cycloartenol caused the depression at a significance level (p 0.01), while the other control 15 drugs cyclobranol, 24-methylenecycloartanol, and y-orizanol showed tendencies to the raise or the depression but insignificantly. In contrast, the compounds of Examples 114, 116, 118, 140, 146, 167, 173, 189, 177, 205, 212, and 213 raised HDL-C levels at high significance levels (p 0.001) and the compounds of Examples 120, 130, 183, 191, 197, 171, 178, 195, 113, and 117 increased at significance Slevels (p 0.01). The compound of Example 170 caused the rise at a low significance level 0.05).

In particular, the compounds of Examples 116, 118, 140, 167, 173, 170, 178, 195, and 213 caused remarkable rises in the HDL-C contents in comparison

I

L ~.II u L..

50 I 4

SI

IIdb II I t 4* 4 44 4 1 with that in the group given the normal diet.

As to the AI, all the compounds tended evidently to depress the AI levels without exception according to method A or B.

The TG, according to method A, was not depressed at the significance level but unvaried or slightly depressed by any compound tested, except that the depression was caused by the compound of Example 49 at a medium significance level (p 0.01) and by the compounds of Examples 100-1, 100-2, and 101 at low significance levels (p 0.05). According to method B, compounds of the present invention as well as control drugs showed tendencies to slight or some degree depression of the TG but insignificantly.

15 As to the PL, according to method A, the compounds of Examples 62, 71, 27, 39, 8, 41, 72, 24, 20, 19, 43, 25, 51, 93, 49, and 100-2 depressed the PL levels at significance levels (p 0.001), the compounds of Examples 29, 37, 60, 74, 83, 58, 13, 14, 100, and 101 at significance levels (p 0.01), and the compounds of Examples 86', 32, 10, 23, 100-1, 105, and 109 at significance levels (p 0.05). The compounds of Examples 2, 82, 12, 16 and 47 showed apparent tendencies to the depression but insignifi- 25 cantly.

According to method B, the control drugs showed tendencies to slight depression of the PL but -51- 1 insignificantly. In contrast, the compounds of Examples 114, 116, 118, 140, 146, 171, 195, and 213 depressed the PL levels at significance levels (p 0.001), the compounds of Examples 120, 130, 189, 205, 212, 191, 197, 170, 178, 113, and 117 at significance levels (p 0.01), and the compounds of Examples 167, 173, 177, and 183 at significance levels (p 0.05).

As to LPO, according to method A, the compounds of Examples 10, 58, 13, 14, 19, 43, 100, 25, 51, 93, 101, 102, 104, and 112 depressed the LPO levels at significance levels (p 0.001), the compound of Example 50 at a significance level, and the compounds O" of Examples 29, 37, 60, 62, 71, 24, 12, 16, 47, 23, 0 15 and 49 significance levels (p 0.05). Other S* compounds of the invention showed distinct tendencies 4*4 to the depression but insignificantly. According to method B, the control drug y-orizanol depressed the LPO at a significance level (p 0.01) and the control S* 20 drugs triterpenyl alcohols showed apparent tendencies to the depression but insignificantly. In contrast, the compounds of Examples 114, 116, 118, 120, 140, 130, 146, 189, 205, 212, 183, 191, 197, 171, 178, I t 195, and 213 depressed the LPO at significance V 25 levels tp 0.001) and the compounds of Examples 167, 173, 177, 170, 113, and 117 at significance levels (p 0.01).

52 1 As described above, most of the compounds according to the present invention, in the tests according to method A or B, tended obviously to raise the HDL-C contents and depress the TC, AI, PL, and LPO levels. In comparison with effects obtained by the administration of the free triterpenyl alcohols alone, it is evident that these hypolipidemic activities of the compounds of the invention are synergistic effects.

Increases of the body weights of rats used for the hypolipidemic activity tests according to method B are shown in Tables 9-1 and 10-1. These tables shows that the body weights of the rats ogiven the normal diet were increased at significance 15 levels (p 0.001) as compared with those of the S0control group rats given the hyperlipidemic diet.

The groups of rats given the hyperlipidemic diet containing each of typical compounds according to the present invention and the gxoups of rats given r 20 the hyperlipidemic diet contaili:n' each of the control t rt drugs showed slight increases in body weight but insignificantly, as compared with the control group of rats given the hyperlipidemic diet alone.

$b; 4* 01,I c, 0sa~ a a C 4 *r fl 1( C r It C ai~e r, Table 1 TC HDL-C TG PL LPO Sample mg/d SD mg/di SD A mg/d, SD mg/d SD n mol/m, SD C 205.1 +27.4 81.5 +10.1 1.517 34.3 +4.8 173.6 +11.0 2.27 +0.38 1 173.5j +32.1 72.0* +9.4 1.410 38.0 +5.2 173.7 +12.1 2.03 +0.23 2 168.0- +30.4 90.3 +9.3 0.860 30.6 +4.9 168.2 +10.8 1.98 +0.27 200.1 +335 76.8 +8.9 1.605 33.3 +6.5 167.0 +13.0 2.40 +0.25 3 200.1 +33.5 76.8 +8.9 1.605 33.3 +6.5 167.0 +13.0 2.40 +0.25 [Notes] In Sample column, C: io Serum of the control group of 10 diet in Tables 1 to 14-1.

rats given the hyperlipidemic 1: Serum of the group of 10 rats given the hyperlipidemic diet containing 1% of cycloartenol as a control drug in Tables 1 to 2 2: Serum of the group of 10 rats given the hyperlipidemic diet containing 1% of cyclobranol as a control drug in Tables 1 to 2.

3: Serum of the group of 10 rats given the hyperlipidemic diet containing 1% of 24-methylenecyloartanol as a control drug in Tables 1 to 2.

Signicicance level (p 0.05) Standard deviation.

These notations are applied to the following tables.

Mark

SD

Cne 4, cyl tO r n 1 r r r r r r Table 2 TC HDL-C AI TG PL LPO Sample Diff. of Diff. of Diff. of Diff of Diff. of 'Diff. of mg/d diff. mg/di diff. mg/di diff. mg/d diff. mg/dk diff. n mol/mk diff.

1 -31.6 -15.4 -9.5 -11.6 -0.10- -7.1 +3.7 +10.9 +0.1 +0.06 -0.24 -10.6 2 -37.1 -18.1 +8.8 +10.8 -0.657 -43.3 -3.7 -10.9 -5.4 -3.1 -0.29 -12.8 3 -5.0 -2.4 -4.7 -5.8 +0.088 +5.8 -1.0 -2.9 -6.6 -3.8 +0.13 +5.8 [Notes] "Diff." means the difference between Q-Qc and of diff." means (Q-Qc) x 100/Qc, wherein Q: Concentration of the component lipid in the sample serum, Qc: Concentration of the component lipid in the serum of the control group.

These notations apply in the following tables.

N CCC C *0 C

CO

O~C

CC

to cJ1 *oe. 0 COG C CCC 0 C~ 0 0 C C 0 0 0 C C to 0 0 CCC C C C 0 C~C C C C C C C 6 0 000

I-,

ca-i coo 0

C

f Table 3 Sample (Example No.)

C

29 37 86 62 74 71 r r r T I I

TOC

MdIn~/~ S D HDL-C AlI mg1df S D

T

mg/dr S D P L P 0 nmol/me S D I. I I 192.6 162.7 161.2 165.6 164.9 159.5 165. 1 161.4 24.3 23.4 22.8 ±25.0 ±24.0 23.2 23.3 23.6 96.2 101.0 110.4 91.9 102.5 108.2 92.4 106.0 8.2 8.2 ±9.4 ±9.7 ±9.2 ±9.3 ±8.9 ±9.4 1.002 0. 611 0.460 0.802 0.609 0.474 0.787 0.523 33.2 32.7 31.0 33.1 32.8 0 33.1 ±8.5 7.8 ±8.3 ±8.8 ±9.2 ±7.4 160.6 155.4 163.2 161.4 156.5 162.1 157.7 14.7 14.9 ±14.7 ±17.2.

±15.2 ±13.8 14.5 15.6 2.33 1.95 1.91 2.06 1.98 2.09 0.35 0.30 ±0.32 ±0.28 0.33 0.32 ±0.33 0.30 32.9 8.3 1.96 [Notes] Mark ~Significance level (p <0.01) Significance level (p 0. 001) Example No.: Serum of the group of 10 rats given the hyperipidemic diet containing 1% of compounds of example number each in Tables 3 to 14-2.

These notations apply in the following tables.

'3 01 4 fl flaw.

0 0~ ho 4.4 t~D 0 flea C 4. 0fl C C C a 0 0,~ C 04* CC C C C *0 C C Table 4 Sample TC HDL-C AI TG PL LPO (Example Diff. of Diff. of Diff. of Diff. of Diff. of Diff. of _mg/i diff. mg/dl diff.. diff. mng/1 diff. mg/d diff. nmol/midiff.

29 -29.9 -15.5 4.8 5.0 -0.391 -39.0 0.5 1.5 -20.3 -11.2 -0.38 -16.3 37 -31.4 -16.3 +14.2 +14.8 -0.542 -54.1 2.2 6.6 -25.5 -14.1 -0.42 -18.0 86 -27.0 -14.0 4.3 4.5 -0.200 -20.0 0.1 0.3 -17.7 9.8 -0.27 -11.6 -27.7 14.4 6.3 6.5 -0.393 -39.2 0.4 1.2 -19.5 -10.'8 -0.35 -15.0 62 -33.1 17.2 +12.0 +12.5 -0.528 -52.7 1.2 3.6 -24.4 -13.5 -0.39 -16.8 74 -27.5 14.3 3.8 4.0 -0.215 -21.5 0.1 0.3 -18.8 -10.4 -0.24 -10.4 71 -31.2 16.2 9.8 +10.2 -0.479 -47.8 0.9 2.8 -23.2 -12.8 -0.37 -15.9 A C on 0 0** 0 0 0 *0 A A 0 @0 *3* A A A o4 0; 8 8 a 4a a oa o 80 8 0 0 0 8 88 8 0 Table Sample T C HDL-C A I T G P L L P. 0 (Example No.) mg/dt SD rg/dt SD mn/dI SD mg/df SD nmol/me SD C 217.1 ±25.2 80.9 ±10.2 1.684 42.8 6.4 179.7 ±10.9 2.31 ±0.40 27 186.1 ±22.7 95.2 ±12.3 0.955 40.4 5.7 160.3 ±11.0 1.99 ±0.38 IKX MX XXXK 39 176.9 ±23.3 98.1 ±11.4 0.803 -9.8 7.2 155.1 ±12.2 1.99 ±0.35 83 192.1 ±23.1 93.3 ±10.8 1.059 41.1 6.5 162.1 9.7 2.02 27 2 194.5 ±23.1 89.2 ±10.7 1.180 41.3 7.1 170.2 ±10.3 2.05 ±0.34 32 190.4 ±23.2 89.2 ±11.2 1.135 40.7 5.5 168.2 ±11.4 2.06 ±0.29 82 195..8 ±25.0 87.9 ±11.3 1.228 42.3 5.3 173.9 ±12.7 2.08 ±0.29 WK X YA. XnX 8 177.8 ±24.5 101.1 ±10.8 0.759 41.3 6.6 148.8 ±11.8 2.01 ±0.29 41 175.6 ±23.8 92.9 ±10.6 0.890 41.5 6.3 156.7 ±10.9 2.01 ±0.35 72 186.5 ±22.7 89.2 ±10.2 1.091 41.8 6.1 160.3 9.8 2.07 ±0.44

V

-V

0*0 0 0 0 0

OP

0 ~p 0 0 0 .4 .40 4 0 0 4 0 0 0 0 0 *4 0 p *60 4 0 Table 6 TC HDL-C AI TG PL LPO Sample Diff. of Diff. of of Diff. of Diff. of Diff. of (Example No.) o /d diff n V/ diff D diff .ng/d diff ng/ diff. nmol/mf diff.

27 -31.0 -14.3 +14.3 +17.7 -0.729 -43.3 2.4 5.5 -19.4 -10.8 -0.32 -14.0 39 -40.2 -18.5 +17.2 +21.2 -0.881 -52.3 3.0 6.9 -24.6 -13.7 -0.32 -14.0 83 -25.0 -11.5 +12.4 +15.3 -0.625 -37.1 1.7 4.0 -17.6 9.8 -0.29 -12.7 2 -22.6 -10.4 8.3 +10.2 -0.504 -29.9 1.5 3.5 9.0 5.0 -0.26 -11.2 32 -26.7 -12.3 8.3 +10.2 -0.549 -32.6 2.1 4.8 -11.5 6.4 -0.25 -10.7 82 -21.3 9.8 7.0 8.7 -0.456 -27.1 0.5 1.2 5.8 3.2 -0.23 9.8 8 -39.3 -18.1 +20.2 +25.0 -0.925 -54.9 1.5 3.5 -30.9 -17.2 -0.30 -13.0 41 -41.5 19.1 +12.0 +14.8 -0.794 -47.1 1.3 3.0 -23.0 -12.8 -0.30 -13.0 72 -30.6 -14.1 8.3 +10.2 -0.593 -35.2 1.0 2-4 -19.4 -10.8 -0.24 -10.4

L

Table 7 Sample T C HDL-C A I T G P L L P 0 (Example No.) mgldt S D ffeldt S D mgldl S D re/di S D nmol/me S D c 166.6 ±14.1 60.2 8.5 1.767 27.2 4.2 136.7 ±12.0 2.53 0.31 146.5 ±15.3 57.6 9.4 1.543 27.3 4.1 123.3 t ±11.9 1. 7737 0.24 58 143.7 ±17.9 62.0 9.6 1.318 26.8 +4.0 121. ]Kx +11.8 1-7 ±-0.13 XX OWN 13 1441 ±16.8 60.5 8.6 1. 388.* 26.2 3.8 122.8 ±12.7 1.94 0.25 14 145.8 ±15.2 64.1 9.2 1.275 25.4 +3.1 120.9 ±11.3 1. 1" ±0.18 V-11- p. 4 40 a a 0 0 a 4 S S 0 4 4 4 0 SOt a 054 4 4 04 0 0 0 0 4 0 5 0 *44 ft a a 4 5 S 0404 o 4 400 0 0 Table 8 r U T Sample (Example

TC

)iff. of ing11di diff.

HDL-C

Diff. of w/,duf f

AI

Df.% of if.diff.

TG

Duff.

nigldl No.) of diff

PL

Diff.

nig/df of di if

LPO

Diff.

,nmo 1/me dif f.

-t i -20.1 -22.9 -22.1 -20.8 -12.1 -13.7 -13.3 -12.5 -2.6 1.8 0.3 3.9 4.3 3.0 0.5 6.5 -0.224 -0.449 -0.379 -0.492 -12.7 -25.4 -21.4 -27.8 0.1 -0.4 -1.0 -1.8 0.4 -1.5 -3.7 -6.6 -13.4 -15.4 -13.9 -15.8 91.8 -11.3 -0.80 -0.71 -0.59 -0.72 -31. 6 -28.1 -23.3 -28.5 I I i L I

~A

pa a if p p a a 0 a a

C

Table 9 Sample T C HDL-C A I T G P L L P O (Example No.) rg/dt SD mg/dt SD mn/dI SD nf/df SD nmol/me SD C 192.2 ±24.0 86.5 9.7 1.232 36.3 5.3 172.6 ±10.5 2.24 ±0.26 24 164.3 ±23.1 94.0 9.8 0.748 34.7 6.4 133.8 1±10.1 1.93 ±+-0.30 162.4 ±22.4 95.7 ±10.4 0.697 34.7 5.1 138.8 ±11.3 1.87 ±0.31 168.8 ±24.5 90.4 ±10.8 0.867 35.2 5.8 143.3 ±11.4 2.02 ±0.35 19 156.1 ±24.5 91.9 9.4 0.699 31.6 5.6 153.6 ±10.2 1.63 ±0.21 43 157.4 ±23.6 91.9 9.6 0.713 33.3 6.4 146.0 ±10.1 1.54 ±0.24 ON ONX KXXo 100 159.1 ±22.6 90.1 ±10.3 0.766 34.3 6.2 155.6 ±11.4 1.68 ±0.23 157.4 ±23.4 92.3 9.6 0.705 34.0 5.4 144.6 ±11.5 1.67 ±0.30 OW* 0 NO W 51 156.5 ±23.5 96.4 9.8 0.623 34.7 4.9 146.0 ±12.0 1.69 ±0.27 93 161.1 23.3 91.5 9.5 0.761 35.1 5.0 150.2 11.0 1.70 0.28XXX 93 161.1T ±23.3 91.5 9.5 0.761 35.1 5.0 150700 ±11.0 1.70 +-0.28

I

2; .09 4' 4'* a e* a aS a 0 4' *0 0e 0 a a 4' 00 6 0 9 a a a 9 4' a a a 4' a a a a a C a Table TC HDL-C AI TG PL LPO (Example No.) Diff. of Diff. of Diff. of Diff. of Diff. of Diff. of m( N /d diff r/g d f diff. diff, wm/ diff.rm/di diff.nmol/me diff.

24 -27.9 -14.5 7.5 8.7 -0.474 -38.8 1.6 4.3 -38.8 -22.5 -0.31 -13.7 -29.8 -15.5- 9.2 +10.6 -0.525 -43.0 1.6- 4.3 -33.8 -19.6 -0.37 -16.7 -23.4 -12.2 3.9 4.5 -0.355 -29.1 1.1 3.0 -29.3 -17.0 -0.22 9.7 19 -36.1 -18.8 5.4 6.2 -0.523 -42.8 4.7 -12.9 -19.0 -11.0 -0.61 -27.2 43 -34.8 -18.1 5.4 6.2 -0.509 -41.7 3.0 8.2 -26.6 -15.4 -0.70 -31.3 100 -33.1 -17.2 3.6 4.2 -0.456 -37.3 -2.0 5.5 -17.6 -10.2 -0.56 -25.2 -34.8 -18.1 5.8 6.7 -0.517 -42.3 -2.3 6.2 -28.0 -16.2 -0.57 -25.6 51 -35.7 -18.6 9.9 +11.5 -0.599 -49.0 1.6 4.3 -26.6 -15.4 -0.55 -24.4 93 -31.1 -16.2 5.0 5.8 -0.461 -37.7 1.2 3.3 -22.4 -13.0 -0.54 -24.3 p 0 0.4 *t4 0 a #0 C 0 0 C a a a 4 t0 4 C 00 4 000 C 0 c-fl Table 11 Sample (Example No.)

C

12 16 47 23 49

T

ng/dLt

HDL-C

we/df SD AlI T

SDI

P, L ff/ctt SD LPO0 nmol/me S D S D t I i- I 167.1 150.4 150.2 150.1 147.6 132.7 ±16.4 15.2 ±15.3 ±15.4 ±15.0 17.0 58.7 58.5 59.4 53.4 58.9 58.0 1.847 1.571 1.529 1.811 1.506 1.288 24.8 24.0 22.0 20.4 20.2 18.3 148.4 140.4 140.1 140.3

EK

133.1 128.4 13.6 12.8 12.7 12.3 ±+12.6 13.0 2.34 2.03 2.04 2.03 2.06 2.02 0.29 +0.22 10.23 0.22 +0.27 ±0.26 *4

S

4*.

a a, Table 12 Sample (Example No.)

TC

Diff.

gr/~Lt of diff.

HDL-C

Diff. of Irg/d, diff.

AI

Diff. of diff

TG

Diff. Of ig/dI diff.

JPL

Dif f. of ng/cC dif f

LPO

Diff. of nrol/mediff 12 -16.7 -10.0 0.2 0.3 -0.276 -14.9 0.8 3.2 8.0 5.4 -0.31 -13.2 16 -16.9 -10.1 0.7 1.2 -0.318 -17.2 2.8 -11.3 8.3 5.6 -0.30 -12.8 47-17.0 -10.2 5.3 9.0 -0.036 1.9 4.4 -17.7 8.1 5.5 -0.31 -13.2 23 -19.5 -11.7 0.2 0.3 -0.341 -18.5 4.6 -18.5 -15.3 -10.3 -0.28 -12.0 49 -34.4 -20.6 0.7 1.2 -0.559 -30.3 6.5 -26.2 -20.0 -13.5 -0.32 -13.7 S 00 00 9 9 4 o 000 9 9 9 .0 0 0 *9 a 00 0 9 0 *0 O 9 04,0 ce oz 0 0 Table 13

LPO

Sample (Example No.)

C

100-1 10C-2 100-3

HDL-C

rrg/ct S D AlI T G me/d S D P !L rrgldt nmol/md S D S D S D 205.1 174.3 166.7 176.6 27.4 ±30.4 ±28.1 ±32.3 81.5 79.1 94.1 73.8 ±10.1 9.4 9.3 10.3 1.517 1.204 0.772 1.393 34.3 28.3 21.6 35.1 173.6 150.5 143.0 152.9 10.6 10.6 2.27 1.51 1.59 1.89 0.38 0.30 0.33 0.26 nnc, 0 C) ('I Table 14 Sample TC HDL-C AI TG PL LPO (Example NSo.) Diff. of Diff. of Diff. of Diff. of Diff. t% o-FiDiff. of mg/dk. diff. -ng/dP. diff. diff rng/dk diff. mg//d k di ff .n mol/Mdjf f 100-1 -30.8 -15.0 2.4 3.0 -0.313 -20.6 6.0 -17.6 -23.1 -13.3 -0.76 -33.5 100-2 -38.4 -18.71+15.1 +18.5 -0.745 -49.1 -12.7 -37.0 -30.6 -17.6 -0.68 -29.8 100-3 -28.5 -13.9 7.7 9.4 -0.124 8.2 0.8 2.3 -20.7 -11.9 -0.38 -16.7 ~L 4 ,n '7 Table 1-1 Sample

C

1 2 3 T C wg/di SD 307.3 ±95.9 190A4 ±51.3 191.5 ±-46.8 191.2 ±-148.4 H1 D L, C ing/df SD 13.5 3.8 10.3 1.4 Al I T G J ig/dl SD 21.8 I56.5 ±20.6 17.5 I55.1 ±12.4 12.0 I53.4 ±17.4 14.8 I53.3 ±19.2 P L L PO 0 Ing/di SD Inmol/inQ SD 142.5 ±30.9 I2.80 ±0.38 140.2 ±40.2 I2.68 ±0.45 138.6 ±38.2 I2.60 ±0.40 133.3 ±25.2 I2.71 ±0.32 1 15.0 +±5.0 12.8 3.2 (Notes] In Sample column, C: Serum of the control group of 16 rats given the hyperlipidemic diet in Tables 1-1 to 10-1.

1: Serum ot the group of 8 rats given the hyperlipidemic diet containing 1% of cycloartenol as a control drug in Tables 1-1 to 4-2 and 9-1.

2: Serum of the group of 8 rats given the hyperlipidemic diet containing 1% of cyclobranol as a control drug in Tables 1-1 to 4-2 and 9-1.

3: Serum of the group of 8 rats given the hyperlipidemic diet containing 1% of 24-methylencycloartanol as a control drug in Tables 1-1 to 4-2 and 9-1.

4. 4, 4, 4 Table 2-1 (Example No.) Kiff. of Diff. of Diff. of Diff. of Diff. of Diff. of w/42 diff. rie/d-P diff. diff. -r,/cld diff. rng/de diff. nmo1/mgdiff-I 1 -117.2 -38.1 13.2 -23.7 -4.3 -19.7 I-1.4 2.5 12.3 1.6 1-0.12 4.3 2 1-115.8 -37.7 I+ 1.5 +11. 1 I-9.8 -45.0 I-3.1 5.5 I-3.9 2.7 1-0.20 7.1 3 1-116.1 -37.8 -0.7 5.2 -7.0 -32.1 -3.2 5.7 19.2 6.5 1-0.09 3.2 .~iA2.

.9, 7

V

Table 3-1 Sample I T C Inag/d SD C 307.3 95.9 N 43.4 8.3 1 202.4 48.3 2 197.5 45.7 3 198.3 45.6 4 190.3 45.0 Example 114 1 138.2 40.3 116 140.1 38.3 118 139.5 39.3 120 203.2 47.4 140 197.4 45.2 130 201.3 44.2 146 19'0.4 47.5

T

4- HDL-C I 13.5 3.81 29.9 6.71 10.3 1.41 15.2 5.61 15.1 5.01 15.3 4.81 24.2 5.6 61.4 16.4 A; IM 47.4 17.01 24.6 8.71 54.8 17.01 2 ±6I 21.0 6.8! A I 21.76 0.452 18.65 11.99 12. 13 11.44 4.711 1. 282 1.943 7.260 2.602 8.586 7. 479 T G n,/di S D 56.5 ±20.6 54.0 +12.1 54.2 14.4 53.3 13.6 54.2 15.2 52.8 18.4 39.0 14.6 54.0 19.0 51.4 19.4 53.0 18.0 52.4 18.4 53.4 18.0 51.4 17.4 140.1 67.3 127.6 130.3 12804 127.6 0 am 96.7 W 99 97.4 98.3 105.3 98.0l 107.2 97.6 28.5 P L I L P. 0 i/f S D I nmol/nf S D 2.82 0.40 10.0 25.8 25.2 24.8 26.0 20.0 19. 1 16.9 20. 1 17.3 19.0 1.82 2.62 2.60 2.58 2.25 R WX 2.04 2WE 2"o9 2.11 WE W ra 2.08 2 WA 2.06 0 WO 2.02 2.07 0.25 0.35 0.32 0.34 0.35 0.22 0.20 0.18 0.20 0.21 0.23 0 39 23.4 5.8 18 2 2.02 0.20 [Notes] N: Serum of the group of 8 rats given the ordinary diet.

The same applies in the following tables.

4: Serum of the group of 8 rats given the hyperlipidemic diet containing 1% of y-orizanol.

Example N i. 3erum of the group of 8 rats given the hyperlipidemic diet contraining 1% of compounds of example numbers each in Tables 3-1 to 10-1.

These notations are applied to the following tables.

r t, Table 4-1 1 I Sample

N

1 2 3 4 Example

TC

Diff. of mgldt diff.

-263.9 -85.9 -104.9 -34.1 -109.8 -35.7 -109.0 -35.5 -117.0 -38.1 -169.1 -55.0 -167.2 -54.4 -168.2 -S4.6 -104.1 -33.9 -109.9 -35.8 -106.0 -34.5 -108.9 -38.1 Diff. of I Diff. of rng/dk. diff.

+16.4 +121.5 3.2 -23.7 1.7 +12.6 1.6 +11.9 1.8 +13.3 +10.7 +79.3 +47.9 +354.8 +33.9 +251.1 +11. 1 +82.2 +41.3 +305.9 7.5 +55.6 dif f -21.31 -97.9 3.11 9.77 9.63 -10.32 -17.05 -20. 48 -19.82 -14.5 -19.16 -13.17 -14.3 -44.9 -44.3 -47.4 -78.4 -94. 1 -91.9 -66.6 -88.0 -60.5

HDL-C

TG

Diff. of mg/dk diff.

-2.5 4.4 -2.3 4.1 -3.2 5.7 -2.3 4.1 3.7 6.5 -17.5 -31.0 6. 1 -10.8 5.1 9.0 3.5 6.2 4.1 7.3 3.1 5.5 5.1 9.0 IDiff nig/di -72.: -12.

-12.

-43 -42 -41.: -34 -42 -32.

PL

%of Y, diff.

8 -52.0 5 8.9 8 7.0 7 -8.4 5 -8.9 4 -31.0 7 -30.5 8 -29.8 8 -24.8 1 -30.0 9 -23.5 T LPO Diff. of n inol/niL. diff I-1.0 -35.5 I I-0.2 7.1 I -0.22 7.8 -0.24 I-0.57 -20.2I -0.78 -27.7I I-0.71 -25.2I -0.74 -26.2I I-0.76 -27.0I -0.80 -28.4I 1'-0.75 -26.6 1 -28.4 1 9.9 +73.3 -14.28 -65.6 -42.5 -30.3 -r 0

N

S S a a

S

4,, .4 #4 0 01 Table 5-1 SaIl *T C Samle uvdt S D c 364.6 ±113.8 N 1 7' 1~ 1 q A 9 19.6 ±5.4 Example 167 173 189 177 205 212 183 191 197 271.0 272.8 *0N 179.5± as 245.5+ !11 MR 19 2.4+ N MM 19 5.0+ 244.0+ 18 8.4+ 2 TN 221.6 43.4 40.5 45.5 43.0 41.3 41.9 42.0 32.6 38.5 M AW 39.7 79.6 At NW 58.4 w NR 35.4 W NW 32.5 37.4 N "W 29.2 28.4 27. 1M ±9.3 ±16.5 ±9.0 ±6.0 4 ±9.0 8.5 ±8.0~ ±6.41 A I 17.60 0.842 2. 405 3.671 4.071 6.554 4.144 4.242 7.356 5.634 7. 177

TOG

Mgd S D 71. 4 29. 5 71.5 19. 4 67. 1 18.6 65.2 27.5 58.4 17.5 63.1 26.4 60.3 21.2 59.7 18.9 65.2 22.5 60.4 23.4 61:'2 24.4~

P

dtc~ 158.7w 90. 3

M

126. 4 127.27 115.0 125.8 113.1 112. 9 125.7 117.3

L

S DI ±31. 7 ±9.41 ±16.4 ±16.3 ±15.61 ±16.51 ±17.01 ±17.21 ±18.11 16.91 N NIK 1.75

NEW

2.28

MW

2.23 WN VW 2.05

NW

2.29

AM

2.02 N, NW 2.01 A NW, 2.08 N NW 2.08 N NEW 2.05 L P 0 !nmo I/ mf S D 2.84 ±0.41 0. 0.29 0.28 0.22 0.30 ±0.21 ±0.22 ±0.23 ±0.21 115.5 14.9 'A1 i Table 6-1 -71 T C HDL-C A I T G P L L P O Sample Diff. Z of Diff. of Diff. of Diff. 2 of Diff. 2 of Diff. of mg/dt diff. mg/dt diff. diff. mg/dt diff. mg/di diff. n mol/mb diff.

N -291.5 -79.9 +20.1 +102.6 -16.8 -95.2 0.1 0.1 -68.4 -43.1 -1.09 -38.4 Example 167. 93.6 -25.7 +60.0 4306.1 -15.2 -86.3 4.3 6.0 -32.3 -20.4 -0.56 -19.7 173 81.8 -22.4 +38.8 +198.0 -13.9 -79.1 6.2 8.7 -31.5 -19.8 -0.61 -21.5 189 -185.1 -50.8 +15.8 80.6 -13.5 -76.9 -13.0 -18.2 -43.7 -27.5 -0.79 -27.8 177 -119.1 -32.7 +12.9 65.8 -11.0 -62.8 8.3 -11.6 -32.9 -20.7 -0.55 -19.4 205 -172.2 -47.2 +17.8 90.8 -13.5 -76.7 -11.1 -15.5 -45.6 -28.7 -0.82 -28.9 212 -169.6 -46.5 +17.6 89.8 -13.4 -75.9 -11.7 -16.4 -45.8 -28.9 -0.83 -29.2 183 -120.6 -33.1 9.6 49.0 -10.2 -58.2 6.2 8.7 -33.0 -20.8 -0.76 -26.8 191 -176.2 -48.3 8.8 44.9 -12.0 -68.0 -11.0 -15.4 -41.4 -26.1 -0.76 -26.8 197 -143.0 -39.2 7.5 4 38.3 -10.4 -59.2 -10.2 -14.3 -43.2 -27.2 -0.79 -27.8

V

A

4 Tak le 7-1 Sample T C HDL C A I *T P L L P0 of q/id S D q/i S D a/i S D /t S D nmol/-e S D C 418.4 t 176.2 16.0 6.1 25.15 81.3 t 46.0 176.8 52.1 3.47 0.34 N 53.0 7.3 27.4 4.1 0.934 53.2 21.1 94.4 12.2 1.15 0.12 Example 171 196.8 51.7 24.2 7.3 7.132 65.6 21.6 118.4 20.4 2.22 0.18 170 261.4 .7 35.7 18.4 6.322 63.5 15.6 125.7 23.8 2.98 0.28 178 191.5 52.8 50.4 21.1 2.800 58.5 12.7 127.0 23.2 2.20 0.25 n ir mm 195 195.3 51.9 49.7 19.7 2.930 57.8 12.4 116.4 12.3 2.18 0.20 213 189.4 55.5 68.4 15.4 1.769 57.6 12.1 115.3 12.5 2.1 0.19 113 253.7 71.8 24.9 8.9 9.189 60.4 20.1 124.3 23.4 3.05 0.30 117 260.5 74.2 26.1 9.2 8.981 63.7 21.0 126.8 24.0 2.62 0.24

I

Table 8-1 Sample

N

pSamp 171 170 178 195 213 113 117 TG HDL-C Diff. of Diff. of mg/di diff. mg/dk diff.

-365.4 8.7 +11.4 71.3 -221.6 -53.0 8.2 51.3 -157.0 -37.5 +19.7 +123.1 -226.9 -54.2 +34.4 +215.0 I -223.1 -53.3 +33.7 +210.6 -229.0 -54.7 +52.4 +327.5 -164.7 -39.4 8.9 55.6 -157.9 -37.7 I +10.1 63.1 LJ1

AI

Diff. of diff.

-t -24.22 -96.3 -18.02 -71.6 -18.83 -74.9 -22.35 -88.9 -22.22 -88.3 -23.38 -93.0 -15.96 -63.5 -16.17 -64.3

TG

Diff. of mg/dt diff.

-28.1 -34.6 -i5.7 -19.3 -17.8 -21.9 -22.8 -28.0 -23.5 -28.9 -23.7 -29.2 -20.9 -25.7 -17.6 -21.6

PL

Diff. of mg/dY diff.

-82.4 -46.6 -58.4 -33.0 -51.1 -28.9 -49.8 -28.2 -60.4 -34.2 -61.5 -34.8 -52.5 -29.7 -50.0 -28.3

LPO

Diff. of n nol/my diff._ -2.32 -66.9 -1.25 -36.0 -0.49 -14.1 S-1.27 -36.6 -1.29 -37.2 S-1.35 -38.9 -0.42 -12.1 -0.85 -24.5 a

N)

U'

C)

a S C a a a a 0*4 0 a

I-

C"

Table 9-1 Sample

C

1 2 3 4 Example 114 116 118 120 140 130 146 Body weight during feed period (g SD) Body weight just before blood sampling (g S D) 1st day 105 4.-0 106 3.9 105 3.6 104 3. 7 106 4. 2 8th day 143 9.0

WX

1 0 2 5.5 144 9.8 147 8.4 147 9.6 147 8.1 145 9.5 146 10.0 147 ±t 9. 2 144 ±9.4 147 ±9.5 144 9.3 149 ±11.0 15th day 181 20.8 26± 9.0 191 18.0 191 12.8 192 16.2 191 12.8 194 18.0 193 17.5 192 17.8 191 18.2 194 17.2 194 18.1 193 17.5 22nd day 231 21. 8 28th day 266 ±t 22.2 284 241 240 240 241 247 244 243 242 246 245 14.7 20.4 16.9 16.7 16.8 16.6 15.8 15.4 15. 7 15.8 15.7 319 272 268 267 269 277 273 274 272 273 271 15.2 20.7 20.4 13.2 13.4 14.9 20.4 18.4 20.1 18.7 18.0 29th day 252 21.7 1 E W 297 17.4 258 20.2 258 20.4 258 19.2 258 18.4 265 14.5 260 19.0 259 20.4 258 23.0 261 15.6 258 20.2 106 5. 9 243 15.8 271 20.2 258 19.1 a a a fleas... a S S a e.g S 01 Table 10-1 Sample

C

N

Example 167 173 189 177 205 212 183 191 Body weight during feed period (g SD) Body weight just before blood sampling (9-±SD)

I-

1st day 105 4.0 105 34 105 3.4 105 3.6 104 3.8 104 3.9 105 3.7 104 3.3 105 3.9 104 3.7 8th day 144 9.5 163 5.8 143 10.4 142 10.5 142 8.1 142 9.6 142 9.8.

143 9.2 142 8-8 142 9.6 142 8.4 15th day 182 20. 9 22nd day 28thiday 233 22.4 268 22. 3 227 191± 190± 187± 188± 190± 189± 188± 8.9 18.4 18.2 12.6 12.8 13.2 18.6 11.9 12.9 286, 241 240 239 14.0 21.7 23.5 20.6 32 2' 2' 238 21.2 21 238 19.8 I 21 239 ±15.8 I 21 239 ±16.5 2 239 U 14.9 2 239 ±16.2 2 ~0± 72± 71± 38± 67± 67± 68± 68± 68± 14.8 20.8 24.2 22.7 25.2 20.4 21.4 20.9 21.7 29th day 253 22.0 29 16.8 259 20.6 258 23.6 256 20.2 256 27.1 256 19.2 257 19.0 257 18.8 257 20.6 257 20.4 188 13. 6 68 20. 8 ~1 C C a C a C 0 a C ate a a Table 14-1 F I Sample I T C I HDL-C A I T G I P L I L P O (Example No. )I /D g S D I SDD mg/d S D nmol/m SDI 1 I I I I c 1326.2 101.4 36.9 8.0 7.840 1 34.0 6.4 165.6 ±13.9 2.80 ±0.421 I NI I I I 1 1 3I 1 101 1216.6 ±63.3 46.4 7.5 3.668 I 28.2 5.7 150.7 ±15.7 2.12 ±0.25 I XX I X I N I Iix.K 102 1205.0 ±58.4 48.1 7.2) 3.262 I 26.7 5.9 140.9 ±11.2 2.02 ±0.18 I K .1 I I I AIxx 104 1204.8 ±57.81 48.7 7.3 3.205 1 27.4 6.2 143.7 ±12.0 2.01 ±0.20 108 1215.6 +59.51 46.6 9.0 3.627 1 29.0 6.6 151.4 ±15.4 2.20 ±0.18 11 I I I I 1 KI x 112 1216.4 ±60.41 46.8 8.51 -3.624 I 28.5 6.3 150.4 ±15.7 I 2.16 ±0.20 1 L I I i II.64- 604 4 4 4 4 ~0 0R~ C *00 see .4 OA- 0 01n Table 14-2 I I 101 102 104 108 112 Diff.

I-109.6 -121.2 1-121.4 I-110.6 1 -109.8

HDL-C

of Ji ff. of diff Img/d2 diff.

Dif f

AT

%of di ff.

TG

)iff.

rng/d, -33.6& -37.2 -37.2 -33.9 -33.7 9.5 +11.2 9.7 9.9 +25.7 +30.4 +32.0 +26.3 +26.8 -4.172 -4.578 635 4. 213 -4.216 -53.2 -58.4 -59. 1 -53.7 -53.8 5.8 7.3 6.6 5.0 5.5 of diff.

-17.1 -21.5 19.4 -14.7 -16.2

PL

)iff. of mg/d2 diff.

LPO

Diff. of nmol/Ine diff -14.9 -24.7 -21.9 -14.2 -15.2 9.0 -14.9 -13.2 8.6 9.2 -0.68 -0.78 -0.79 -0.60 -0.64 -24.3 -27.9 -28.2 -21.4 -22.9 J 77 1 In the anti-hyperlipidemic assay according to method A, the daily dosage of each compound according to the present invention was 1% of 10 g/day of the hyperlipidemic diet, i.e. 100 mg. For instance, cyclobranyl esters of and paminobenzoic acid (Examples 49, 47, and 43), "Iyotrany-L ester of linoleic acid (Example 51), and cyclobranyl ester of nicotinic acid (Example (100 mg each) contain the bound and p-aminobenzoic acids (24.5 mg each), linoleic acid (39.9 mg), and nicotinic acid (22.6 mg), respectively.

*1 These organic acids in free form were added each to the hyperlipidemic diet, and the resulting diets (10 g/day each) were administered to rats in 1 15 the same manner as in the above anti-hyperlipidemic assay, but the anti-hyperlipidemic effect was not observed with the above doses (22.6 39.9 mg/day) of the free acids. Thus, it has been confirmed that the effect of the compounds of Examples 49, 47, 43, 20 51, and 50 is not attributable to the or S, p-aminobenzoic acid, linoleic acid, or nicotinic acid combining with cyclobranol.

72 K In method B, the dosage of each compound or according to the present invention is roughtly calculated as 210 mg/day for each rat at most from the amount of the fed diet containing the compound.

For example, cycloartenyl ester of 4-hydroxy-3- 78 1 methoxy-a-methylcinnamic acid (Example 114), cyclobranyl ester of 3-ethoxy-4-hydroxy-a-methylcinnamic acid (Example 140), cylobranyl ester of 5-amino-2-methoxybenzoic acid (Example 173), cycloartenyl ester of p-amino-a-methylcinnamic acid (Example 205), and cyclobranonyl ester of m-aminoa-methylcinnamic acid (Example 212) (210 mg each) contain the bound 4-hydroxy-3-methoxy-a-methylcinnamic acid (70.8 mg), 3-ethoxy-4-hydroxy-a-methylcinnamic acid (72.5 mg), 5-amino-2-methoxybenzoic acid (59.4 mg), p-amino-a-methylcinnamic acid (63.4 mg), and m-aminoa-methylcinnamic acid (62.0 mg), respectively.

ec These organic acids in free form were added each to the hyperlipidemic diet, and the resulting diets were administered to rats in the same manner as in the above anti-hyperlipidemic assay, but the anti-hyperlipidemic effect was not observed with the above doses (59.4 72.5 mg/day) of the free acids. Thus, it has been proved that the effect of 20. the compound according to the present invention is ,not attributable to the organic acid liberated by the hydrolysis of the triterpeny ester of organic acid.

Of the compounds according to the present invention, the most favorable as anti-hyperlipidemic agents are esters from combinations of the triterpenyl alcohols with substituted cinnamic, a-(Cl-C 4 alkyl) cinnamic, and benzoic acid which have one or two 79 1 substituents on the benzene ring, the one substituent being a hydroxyl or amino group and the two being C1-C 4 alkoxy and hydroxyl groups or C -C 4 alkoxy and amino groups.

For clinical use, the compounds of the invention can be administered parenterally or preferably orally. Suitable forms of the compounds for oral dosage are of tablets (uncoated or coated with sugar or the like), granules, powders, coated tablets, sugar-coated tablets, capsules, emulsions, etc. which additionally contain pharmaceutically acceptable carriers. For example, the carriers t «include lactose, white sugar, mannitol, anhydrous o dextrose, starch, sorbitol, glycine, potassium 15 phosphate, and microcrystalline cellulose as excipients; starch, gelatin, gum arabic, anhydrous dextrose, white sugar, sorbitol, mannitol, traganth, hydroxypropylcellulose, hydroxypropoxymethylcellulose, carboxymethylcellulose, methacrylic acid-methyl acrylate copolymer, polyvinylpyrrolidone, and sodium aliginate as binders; t v stearic acid, hardened oil, magnesium stearate, calcium stearate, polyoxyethylene monostearate, talc, silicon oxide, and polyethylene glycol as lubricants; potato starch and starch containing a surfactant or the like, as disintegrating agents; and sodium 80 1 laurylsulfate as a wetting agent. For parenteral dosage, the present compounds can be used in the form of intramuscular-injectable or suppository composition.

Base materials for the suppository include cacao butter, Witepsol, Subanal, polyethylene glycol, polypropylene glycol, glycerogelatin, gelatin capsules, etc. In addition, the suppository may contain a known safety preservative such as methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, or butylhydroxyanisole, and pharmaceutically acceptable coloring matter.

While depending upon the mode of administration, the age, weight, and conditions of the patient, and the kind of disease, daily doses of the present

I

15 compounds for man are generally from 0.01 to 5 g, preferably from 0.02 to 1.5 g.

Triterpenyl alcohols for the novel esters of the present invention include also lanosterol, lanostenol, agnosterol, cyclosadol (38-hydroxy-24methylene-9,19-cylo-93-lanosta-23-ene), dihydroagno- 11terol, cyclolaudenol, cycloartanol, cycloeucalenol, euphol, butyrospermol, tirucallol, euphorbol, and Ct dammerrdienol, besides cycloartenol, cyclobranol, O 0* and 24-methylenecycloartanol mentioned above.

Further, anti-hyperlipidemic effect can also be expected for the esters of the organic acids, i defined above, combined with sterols, e.g. dihydro-

L

81 1 -sitosterol, dihydro-y-sitosterol, campesterol, 1-sitosterol, y-sitosterol, stigmasterol, 24iethylenecholesterol, episterol, and 22-dihydroergost -rol, which are analogous in structure to triterpenyl alcohols.

t t 82J 1 Example 1 Preparation of cycloartenyl-3,4-diacetoxycinnamate Toluene (20 ml) was added to 3,4-diacetoxycinnamic acid (4.65 g, 0.018 mole) and cooled to 0°C.

Thionyl chloride (10.0 ml, 10 equivalents) was added dropwise thereto and further pyridine (0.5 ml) was added. The reaction mixture was conducted at 60 0 C for 3 hours. Then, the resulting mixture was evaporated to dryness under reduced pressure and cycloartenol (5.0 g, 0.012 mole) and pyrydine (50 ml) were added thereto and the mixture was stirred at 60 0 C for 1 hour.

Thereafter, the solvent was removed by distillation under diminished pressure. The residue was dissolved 'in chloroform, and the solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with chloroform (3x50 ml). The combined chloroform layer was dried, concentrated under reduced pressure, and purified by silica gel column chromatography (solvent:ethyl acetate-hexane giving cycloartenyl-3,4-diacetoxycinnamate S" (5.6 g) in a 71 yeild, m.p. 125.5 126.5 0

C.

24 Specific rotation [a] 2 4 +36.60 (C 1.00, CHC1) Analysis, Calcd. for C 43

H

60 0 6 672.91): SC, 76.75; H, 8.99. Found: C, 76.82; H, 9.04.

IRY, KBr (cm 2930, 2860, 1773, 1710, 1637, 1502, 1370, 1257, 1205, 1176 PMR (CDCI 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 1 0.60 (1H, 1/2 ABq, 4.2 Hz), 0.70-2.40 (27H, m), 0.90 (6H, 0.96 (6H, 1.61 (3H, bs), 1.68 (3H, bs) 2.31 (6H, 4.40-5.40 (2H, m), 6.40 (1H, 1/2 ABq, 16 Hz), 7.00-7.60 (3H, m), 7.58 (1H, 1/2 ABq, 16 Hz) Example 2 Preparation of cycloartenyl-3,4-dihydroxycinnamate Dioxane (540 ml) was added to cycloartenyl-3,4diactoxycinnamate (27.0 g, 0.040 mole) prepared according to the procedure of Example 1. To the mixture cooled to 0°C was added dropwise 25 aqueous ammonia (27 ml), and this reaction mixture was stirred at for 1 hour. Then the resulting mixture was evaporated to dryness under reduced pressure to give crude crystals, which were then washed with water and recrystallized from acetone-water thereby giving cycloartenyl-3,4-dihydroxycinnamate (21.1 g) in a 89 yield m.p. 230 231 0

C.

23 Specific rotation [a]D +44.70 (C 0.19, CHC1 3 Analysis, Calcd.for C 3 9

H

5 6 0 4 588.84): C, 79.54; H, 9.59. Found: C, 79.62; H, 9.52.

IRv, KBr 3470, 3300, 2910, 2850, 1680, 1602, 1525, 1440, 1275, 1180, 972 S! PMR (CDC1 3 -DMSO-d 6 0.20-0.70 (2H, 0.40-2.40 i 25 (27H, 0.90 (6H, 0.94 (6H, 1.58 (3H, bs), 1.65 (3H, bs), 4.30-4.80 (1H, m), 4.80-5.30 (1H, 6.17 (1H, 1/2 ABq, 15 Hz), 84 1 6.60-7.20 (2H, 6.99 (1H, bs), 7.40 (1H, 1/2 ABq, 15Hz), 8.87 (1H, bs), 9.25 (1H, bs).

Example 3 Preparation of cycloartenyl-3,4-dipropionyloxycinnamate Toluene (26 ml) and thionyl chloride (34ml, 4.6 equivalents) were added to 3,4-dipropionyloxycinnamic acid (29.0 g) at 0°C. This reaction mixture was stirred at 60 0 C for 20 minutes. Then, the resulting mixture was evaporated to dryness under reduced pressure, and the residue was dissolved again in toluene (50 ml).

To the mixture cooled to 0°C was added a solution of cycloartenol (30.0 g, 0.070 mole) in pyridine (60 ml) and the whole was stirred at 200C for 1 hour. Then, the solvent was removed by distillation under diminished pressure. The resulting residue was dissolved in chloroform, and the'solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with chloroform (3x500 ml). The combined chloroform layer was dried and evaporated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (solvent: methylenechloride), giving cycloartenyl-3,4-dipropionyli I oxycinnamate (35.7 g) in a 72 yield.

Example 4 Preparation of cycloartenyl-3,4-dihydroxycinnamate Dioxane (20 ml) was added to cycloartenyl-3,4dipropionyloxycinnamate (1.0 g, 1.43 mmoles) prepared '*l 8 i 1 according to the procedure of Example 3 and was stirred at 0°C. Thereto was added dropwise 25 aqueous ammonia (2.0 ml) and the whole was stirred at 20 0 C for 1 hour. The resulting mixture was evaporated to dryness under reduced pressure. The crude crystals were washed with water and recrystallized from acetone-water (1:1, V/V) giving cycloartenyl-3,4-dihydroxycinnamate (756 mg) in a 90 yield, m.p. 230 231 0

C.

23 Specific rotation [ajD +44.70 (C 0.19, CHCl 3 Analysis, Calcd. for C39H560 588.84): C, 79.54; H, 9.59. Found: C, 79.59; H, 9.63.

IRV, KBr (cm 1 3470, 3300, 2910, 2850, 1680, 1602, 1525, 1440, 1275, 1180, 972.

PMR (CDCl 3 -DMSO-d 6 0.20-0.70 (2H, 0.40- 2.40 (27H, 0.90 (6H, 0.94 (6H, s), 1.58 (3H, bs), 1.65 (3H, bs), 4.30-4.80 (1H, m), 4.80-5.30 (1H, 6.17 (1H, 1/2 ABq, 15 Hz), 6.60-7.20 (2H, 6.99 (1H, bs), 7.40 (1H, 1/2 ABq, 15 Hz), 8.87 (1H, bs), 9.25 (1H, bs).

Example Preparation of cycloartenyl-p-acetoxycinnamate Toluene (18 ml) was added to p-acetoxycinnamic Sacid (18.1 g, 0.088 mole) and the mixture was cooled Sto 0°C. Thionyl chloride (31 ml, 5 equivalents) and pyridine (1.0 ml) were added dropwise thereto, and the whole was heated at 60 0 C for 15 minutes. After concentration of the resulting mixture, toluene (35 ml) r 1 and pyridine (50 ml) were added, and the whole was cooled to 0 C. Thereto was added dropwise a solution of cycloartenol (25.0 g, 0.059 mole) in pyridine ml). The mixture was refluxed for 40 minutes and then evaporated to dryness under reduced pressure.

The resulting residue was dissolved in chloroform, and the solution was washed with saturated aqueous solution of Sodium bicarbonate. The aqueous layer was extracted with chloroform (3x500 ml). The combined extracts was dried, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (solvent: methylene chloride- 1 hexane, giving cycloartenyl-p-acetoxy- N cinnamate (32.0 g) in a 89 yield, m.p. 153 156 0

C.

4. 21.5 Specific rotation [a]D +42.60 (C 1.02, CHCl 3 Analysis, Calcd.for C 41

H

58 0 4 614.87): C, 80.08; H, 9.51. Found: C, 80.13; H, 9.42.

IRv, KBr 2920, 2850, 1765, 1695, 1500, 1370, 1270, 1195, 1160.

PMR (CDC1 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.60 (1H, 1/2 ABq, 4.2 Hz), 0.70-2.40 (27H, m), 0.90 (6H, 0.96 (6H, 1.60 (3H, bs), 1.67 (3H, bs), 2.30 (3H, 4.50-5.30 (2H, m), 6.40 (1H, 1/2 ABq, 15 Hz), 6.90-7.80 (4H, m), 7.60 (1H, 1/2 ABq, 15 H Example 6 Preparation of cycloartenyl-p-hydroxycinnamate 87 1 Cycloartenyl-p-acetoxycinnamate (28.0 g, 0.046 mole) prepared according to the procedure of Example was dissolved in dioxane (280 ml). Then 25 aqueous ammonia (28 ml) was added dropwise to the solution at 0°C. The reaction mixture was stirred at 50 0 C for 2 hours, and then evaporated to dryness under reduced pressure. The residue was recrystallized from acetonewater giving cycloartenyl-p-hydroxycinnamate (23.4 g) in a 90 yield, m.p. 248 248.5 0

C.

26 Specific rotation [a]D +45.90 (C 0.98, CHC13 D IRV, KBr (cm 3190, 2930, 2850, 1705, 1670, 1605, 1582, 1512, 1440, 1280, 1170, 830.

PMR (CDCl 3 -DMSO-d 6 0.20-0.80 (2H, 0.50- 2.40 (27H, 0.88 (6H, 0.95 (6H, s), 1.57 (3H, bs) 1.64 (3H, bs), 4.30-4.84 (1H, m), 4.84-5.30 (1H, 6.28 (1H, 1/2 ABq, 15 Hz), 6.60-7.00 (2H, m) 7.20-7.60 (2H, m) 7.44 (1H, 1/2 ABq, 15 Hz), 9.75 (1H, bs).

Example 7 Preparation of cycloartenyl-o-acetoxybenzoate Thionyl chloride (39.0 ml, 5 equivalents) and S'pyridine (2.0 ml) were added dropwise to acetylsalicylic I *acid (19.0 g, 0.106 mole) in benzene (95 ml) with C C C t a t stirring at 20 0 C. This reaction mixture was stirred at 500C for 1 hour, and then evaporated to dryness under reduced pressure. Benzene (100 ml) was added thereto and the whole was stirred at 0°C, and cycloartenol m Ill I ,i 88 1 (30.0 g, 0.070 mole) dissolved in pyridine (100 ml) was added and the whole was stirred at 500 for 20 minutes.

After the reaction mixture was concentrated under reduced pressure, the residue was dissolved in methylene chloride, and the solution was washed with saturated aqueous solution of Sodium binarbonate.

The aqueous layer was extracted with methylene chloride (3x500 ml). The combined extracts were dried, evaporated under reduced pressure to remove methylene chloride, and the residue was purified by silica gel column chromatography (solvent: toluene-methylene chloride, giving cycloartenyl-o-acetoxybenzoate (30.9 g) in a 75 yield, m.p. 138- 139 0

C.

re 25 Specific rotation [a]25 +61.40 (C 0.99, CHCl) S 15 Analysis, Calcd.for C 39

H

56 0 4 588.84): C, 79.54; H, 9.59. Found: C, 79.47; H, 9.63.

IRV, KBr 2993, 2850, 1768, 1715, 1605, 1447, 1288, 1260, 1190, 1120.

PMR (CDC1 3 0.38 (1H, 1/2 ABq, 4.2 Hz), 0.59 (1H, 1/2 ABq, 4.2 Hz), 0.70 2.30 (27H, m), t i 0.81 (6H, 0.97 (3H, 1.00 (3H, s), 9 L 1.60 (3H, bs), 1.67 (3H, bs), 2.35 (3H, s), 4.50-5.30 (2H, m) 6.90-8.20 (4H, m).

Example 8 25 Preparation of cycloartenyl-o-hydroxybenzoate Sixty milliliter of 25 aqueous ammonia was added dropwise to cycloartenyl-o-acetoxybenzoate (30.0 g, m i 1 0.051 mole) in dioxane (600 ml) with stirring at 0 0

C.

The reaction mixture was stirred at 55 0 C for 4 hours.

Then the solvent was removed by distillation under reduced pressure, the residue was dissolved in methylene chloride, and the solution was washed with saturated aqueous solution of Sodium bicarbonate.

The aqueous layer was extracted with methylene chloride (3x500 ml). The combined extracts were dried and evaporated to dryness under reduced pressure. The residue was recrystallized from methylene chloridemethanol giving cycloartenyl-o-hydroxybenzoate (26.2 g) in a 95 yield, m.p. 132 133 0

C.

25 Specific rotation [D +72.90 (C 1.01, CHC 3 Analysis, Calcd. for C 3 7

H

5 4 0 3 546.80): 15 C, 81.27; H, 9.95. Found: C, 81.36; H, 9.90.

IRV, KBr (cm 3130, 2910, 2850, 1663, 1610, 1480, 1295, 1245, 1210, 1155, 1090, 965, 760.

PMR (CDC1 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.61 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, m), t 20 0.91 (6H, 0.98 (3H, 1.04 (3H, s), t 1.60 (3H, bs), 1.68 (3H, bs), 4.60-5.30 (2H, S 6.60-8.00 (5H, m).

Example 9 Preparation of cycloartenyl-p-acetoxybenzoate I 25 Thionyl chloride (52 ml, 5 equivalents) and Pyridine (1.0 ml) was added to p-acetoxybenzoic acid (25.4 g, 0.141 mole) in benzene (100 ml) with stirring 1 1 at 0VC. The reaction mixture was stirred at 55 0 C for minutes, and evaporated under reduced pressure, then the residue was cooled to 5 0 C. After addition of benzene (200 ml) thereto, cycloartenol (40.0 g, 0.094 mole) dissolved in pyridine (200 ml) was added and the mixture was stirred at 60 0 C for 30 minutes. Then the resulting mixture was concentrated under reduced pressure to remove the solvents. The residue was dissolved in chloroform, and the solution was washed with saturated aqueous solution of Sodium bicarbonate.

The aqueous layer was extracted with chloroform (3x600 ml). The combined chloroform layer was dried and evaporated to dryness. The crude crystals were recrystallized from methylene chloride-methanol V/V) giving cycloartenyl-p-acetoxybenzoate (52.5 g) in a 95 yield, m.p. 141 142 0

C.

25 Specific rotation [aD +58.60 (C 1.00, CHC 3 Analysis, Calcd. for C 3 9

H

5 6 0 4 588.84): C, 79.54; H, 9.59. Found: C, 79.52; H, 9.68.

IRv, KBr (cm 2930, 2850, 1760, 1720, 1600, C I C t 1360, 1272, 1189, 1159, 1120.

S PMR (CDC1 3 0.40 (1H, 1/2 ABq,, 4.2 Hz), 0.61 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, m), I 0.91 (6H, 0.98 (3H, 1.03 (3H, s), 25 1.60 (3H, bs), 1.69 (3H, bs), 2.32 (3H, s), 4.60-5.30 (2H, 6.96-7.40 (2H, m) 7.80- 8.22 (2H, m).

the amount of the fed diet containing the compcund.

For example, cycloartenyl ester of 4-hydroxy-3- 91 1 Example Preparation of cycloartenyl-p-hydroxybenzoate Dioxane (500 ml) was added to cycloartenyl-pacetoxybenzoate (27.0 g, 0.046 mole) prepared according to the procedure of Example 1. Then 25 aqueous ammonia (50 ml) was added dropwise to the solution, and the reaction mixture was stirred at 20 0 C for hours. Then the mixture was evaporated under reduced pressure to remove the solvent. The resulting residue was purified by silica gel column chromatography (solvent: methylene chloride-ethanol, (98:2, giving cycloartenyl-p-hydroxybenzoate containing a 9. molecular ethanol in a 96 yield, m.p. 180 182 0

C.

**Specific rotation [a Srotation +66.10 (C 1.00, CHCl 3 Analysis, Calcd. for C37H 5403 2H5H 592.87): C, 79.00; H, 10.20. Found: C, 79.11; H, 10.14.

IRv, KBr (cm 3450, 3150, 2950, 2850, 1715, 1689, 1612, 1600, 1515, 1310, 1280, 1160.

PMR (CDC 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.60 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, m), 0.91 (6H, 0.97 (3H, 1.03 (3H, s), 1.60 (3H, bs), 1.68 (3H, bs), 4.50-5.30 (2H, 6.60-7.05 (2H, 6.88 (2H, 1/2 ABq, 8.1 Hz), 7.11 (1H, bs), 7.60-8.20 (2H, m), 7.82 (2H, 1/2 ABq, 8.1 Hz).

Example 11 a Preparation of cycloartenyl-m-acetoxybenzoate 92 1 Thionyl Chloride (52 ml, 5 equivalents) and Pyridine (0.5 ml) were added to m-acetoxybenzoic acid (25.0 g, 0.139 mole) in benzene (100 ml) with stirring at 0°C. The reaction mixture was stirred at 60 0 C for 30 minutes and evaporated under reduced pressure.

Benzene (200 ml) was added to the concentrate and the solution was stirred at 0°C. Cycloartenol (40.0 g, 0.094 mole) dissolved in pyridine (200 ml) was added dropwise thereto, and the reaction mixture was stirred at 60 0 C for 1 hour. The resulting mixture was evaporated under reduced pressure, to give a crystalline residue. This residu was dissolved in chloroform, and the solution was washed with saturated aqueous t a .solution of sodium bicarbonate. The aqueous layer 15 was extracted with chloroform (3x500 ml). The combined extracts were dried, concentrated under reduced pressure and the residue was purified by silica gel column chromatography (solvent: methylene chloridehexane, giving cycloartenyl-m-acetoxy- 20 benzoate (49.0 g) in a 89 yield, m.p. 122 123 0

C.

cc Specific rotation []25.5 +60.80 (C,0.99, CHCl 3 3 Analysis, Calcd. for C 39

H

5 6 0 4 588.84): 79.54; H, 9.59. Found: C, 79.60; H, 9.55.

-1 IRV, KBr (cm 2930, 2850, 1769, 1715, 1585, S, 25 1440, 1370, 1230, 1275, 1212.

PMR (CDC1 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.61 (1H, 1/2 ABq, 4-2 Hz), 0.60-2.30 (27H, m), g i 93 1 0.91 (6H, 0.98 (3H, 1.03 (3H, s), 1.60 (3H, bs), 1.68 (3H, bs), 2.32 (3H, s), 4.60-5.30 (2H, 7.10-8.05 (4H, m).

Example 12 Preparation of cycloartenyl-m-hydroxybenzoate Dioxane (400 ml) was added to cycloartenyl-macetoxybenzoate (27.0 g, 0.036 mole) prepared according to the procedure of Example 11. While stirring the mixture at 0 C, 25 aqueous ammonia (40 ml) was added dropwise. Then the reaction mixture was stirred at 0 C fDr 1.5 hours. The resulting mixture was evaporated to dryness under reduced pressure. Then the residual crystals were dissolved in chloroform, a, o and the solution was washed with saturated aqueous S 15 solution of Sodium bicarbonate. The aqueous layer 9 was extracted with chloroform (3x300 ml). The combined extracts were dried and evaporated under reduced pressure, separating out crude crystals which were then recrystallized from methylene chloride-hexane (1:4, 20 giving cycloartenyl-m-hydroxy benzoate (22.7 g) in a 91 yeild. m.p. 176 177.5 0

C.

25.5 a Specific rotation [a] 2 5 +65.40 (C 1.01, CHC1 3 D 3 Analysis Calcd. for C 3 7

H

5 4 0 3 546.80): C, 81.27; H, 9.95. Found: C, 81.21; H, 9.99.

i t 25 IRV, KBr (cm1) 3380, 2950, 2930, 2850, 1710, 1692, 1600, 1450, 1310, 1290, 1110, 970, 758.

PMR (CDC1 3 0.3© (1H, 1/2 ABq, 4.2 Hz), it 94 1 0.60 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, m), 0.90 (6H, 0.97 (3H, 1.02 (3H, s), 1.60 (3H, bs) 1.68 (3H, bs) 4.52-5.30 (2H, m) 6.90-7.70 (4H, m) Example 13 Preparation of cycloartenyl-o-methoxybenzoate Cycloartenol (16.0 g, 0.038 ml) was dissolved in pyridine (160 ml). While the solution was stirred at 0°C, o-methoxybenzoyl chloride (7.0 ml, 1.2 equivalents) was added dropwise thereto and the mixture was allowed to react at 400C for 1 hour. Then the resulting mixture was evaporated under reduced pressure, and the crystalline residue was dissolved in chloroform, and the solution was washed with saturated aqueous t 15 solution of Sodium bicarbonate. The aqueous layer was extracted with chloroform (3x100 ml). The Scombined extracts were dried and evaporated to dryness under reduced pressure. The resulting crude crystals were recrystallized from methylene chloride-methanol giving cycloartenyl-o-methoxybenzoate SI (18.0 g) in a 86 yield. m.p. 141 1420C.

r 25 Specific rotation +47.50 (C 1.02, CHC3) Analysis, Calcd. for C 38

H

56 0 3 560.83): C, 81.38; H, 10.07. Found: C, 81.33; H, 10.15.

(i 25 IRv, KBr (cm 2930, 2850, 1720, 1696, 1598, 1460, 1298, 1250, 1130.

PMR (CDC13)6: 0.38 (1H, 1/2 ABq, 4.2 Hz), i1 1 0.60 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, m), 0.91 (3H, 0.96 (6H, 1.00 (3H, s), 1.60 (3H, bs), 1.67 (3H, bs), 3.88 (3H, s), 4.50-5.30 (2H, 6.70-8.00 (4H, m).

Example 14 Preparation of cycloartenyl-p-methoxybenzoate To a solution of cycloartenol (20.0 g, 0.047 mole) in pyridine (150 ml) cooled to 0°C, there was added dropwise p-methoxybenzoic chloride (9.5 ml, 1.2 equivalents), and the whole was stirred at 50 0 C for 3 hours. The resulting mixture was evaporated to dryness under reduced pressure. The residual crystals

C

F

were dissolved in chloroform, and the solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with Schloroform (3x200 ml). The combined extracts were c dried and evaporated to dryness under reduced pressure.

The crude crystals were recrystallized from methylene chloride-methanol V/V) giving cycloroartenyl-prI 20 methoxybenzoate (25.0 g) in a 95 yield. m.p. 129.5 1300C.

Se i r o Specific rotation [a]D +62.90 (C 0.99, CHC1 3 Analysis, Calcd. for C3 H5603 560.83): 81.38; H, 10.07. Found: C, 81.31; H, 10.15.

IRv, KBr (cm 2910, 2850, 1711, 1605, 1508, 1270, 1250, 1165, 1115, 1100.

PMR (CDC1 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.61 (1H, Sr i V Ia i i II1II 1 I~--CIIIIC III~ 1 1/2 ABq, 4.2 Hz), 0.70-2.30 (27H, 0.91 (6H, 0.98 (3H, 1.03 (3H, 1.60 (3H, bs), 1.68 (3H, bs), 3.84 (3H, 6.70-7.05 (2H, m), 7.80-8.12 (2H, .m) Example Preparation of cycloartenyl-m-methoxybenzoate Thionyl Chloride (24 ml, 5 equivalents) and pyridine (0.5 ml) were added to m-methoxybenzoic acid (9.6 g, 0.063 mole) in benzene (50 ml) with stirring at 0°C. The reaction mixture was stirred at 40 0

C

for 30 minutes. The resulting mixture was evaporated under reduced pressure and benzene (100 ml) was added to the residue. Cycloartenol (18.0 g, 0.042 mole) dissolved in pyridine (100 ml) was added dropwise thereto at 0°C and the whole was stirred at 40 0 C for 1 hour and at 50 0 C for 1 additional hour. Then the solvent was removed by distillation under reduced pressure. The residue was dissolved in chloroform, and the solution was washed with saturated aqueous solution of Sodium bicarbonate. The aqueous layer was extracted with chloroform (3x200 ml). The combined extracts were dried, and evaporated under reduced pressure. The crude crystals were recrystallized from methylene chloride-methanol giving cycloartenyl-m-methoxybenzoate (23.0 g) in an almost quantitative yield. m.p. 127 128 0

C.

25.5 Specific rotation [a]D +63.70 (C 1.00, CHC1 3 1 -1 1 Analysis, Calcd. for C38H5603 560.83): C, 81.38; H, 10.07. Found: C, 81.44; H, 10.01.

IRV, KBr (cm-1) 2930, 2850, 1715, 1700, 1584, 1450, 1285, 1275, 1100, 1045, 755.

PMR (CDC1 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.60 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, 0.91 (6H, 0.98 (3H, 1.04 (3H, 1.60 (3H, bs), 1.68 (3H, bs), 3.84 (3H, 4.60-5.30 (2H, m), 6.90-7.80 (4H, m).

Example 16 Preparation of cycloartenyl-o-nitrobenzoate o-Nitrobenzoic acid (4.7 g, 0.028 mole) was dissolved in dioxane (50 ml). Thionyl chloride (10.0 ml, 5 equivalents) and dimethylformamide (0.2 ml) was added dropwise to the former solution with stirring at r' 0 0 C. The whole was stirred at 50 0 C for 30 minutes.

The resulting mixture was concentrated under reduced pressure to remove the excess thionyl chloride.

Dioxane (50 ml) was added to the residue and the soluc 20 tion was stirred at 0 C. Cycloartenol (10.0 g, 0.023 mole) dissolved in pyridine (50 ml) was added dropwise thereto, and the mixture was stirred at 70 0 C for U minutes. The resulting mixture was concentrated under V t reduced pressure, and dissolved in chloroform (100 ml).

V t t c 1%t 25 The solution was washed with saturated aqueous solution of Sodium bicarbonate. The aqueous layer was extracted with chloroform (3x100 ml). The combined extracts were i i. I i U I r 9°.

1 dried and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (solvent: chloroform-hexane giving cycloartenyl-o-nitrobenzoate (13.0 g) in a 96 yield.

m.p. 166 167 0

C.

22 Specific rotation [a]D +94.80 (C 1.01, CHC1 3 Analysis, Calcd. for C37H5304N 575.80): C, 77.17; H, 9.28; N, 2.43. Found: C, 77.28; H, 9.21; N, 2.42.

IR\, KBr (cm 1 2930, 2850, 1710, 1535, 1375, 1300, 1130.

PMR (CDC1 3 0.38 (1H, 1/2 ABq, 4.2 Hz), 0.59 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, m), 0.89 (3H, 0.91 (3H, s) 0.94 (3H, 0.97 (3H, 1.60 (3H, bs), 1.68 (3H, bs), 4.60- 5.30 (2H, 7.50-8.00 (4H, m) Example 17 Preparation of cycloartenyl-o-aminobenzoate Cycloartenyl-o-nitrobenzoate (20.0 g, 0.035 mole) prepared according to the procecure of Example 16 was suspended in acetic acid (800 ml). By adding zinc powder (20.0 the mixture was stirred at 50 0

C

for 3 hours. Then, zinc powder was removed by filtrae" tion and the filtrate was concentrated under reduced 25 pressure. The residue was dissolved in chloroform, and the solution was washed with saturated aqueous solution of Sodium bicarbonate. The aqueous layer was 1 extracted with chloroform (3x500 ml). The combined extracts were dried, concentrated under reduced pressure and the residue was purified by silica gel chromatography (solvent: methylene chloride-hexane, giving cycloartenyl-o-aminobenzoate (15.0 g) in a 80 yield. m.p. 185 186 0

C.

25.5 Specific rotation [a]D +77.50 (C 1.02, CHC1 3 Analysis, Calcd. for C 37

H

55 0 2 N 545.82): C, 81.41; H, 10.16; N, 2.57. Found: C, 81.48; H, 10.18; N, 2.53.

-1 IRv, KBr 3470, 3350, 2930, 2850, 1670, 1620, 1290, 1248, 1155, 760.

PMR (CDCl 0.40 (1H, 1/2 ABq, 4.2 Hz), 0.61 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, m), 0.91 (6H, 0.97 (3H, 1.03 (3H, s), 1.60 (3H, bs), 1.67 (3H, bs), 4.60-5.32 (2H, m), ti 5.50 (2H, bs), 6.40-8.00 (4H, m).

Example 18 Preparation of cycloartenyl-p-nitrobenzoate t* t t 20 Dioxane (30 ml) was added to p-nitrobenzoic acid (2.95 g, 0.018 mole). Thionyl chloride (6.5 ml, equivalents) and dimethylformamide (0.3 ml) were added dropwise to the former solution at 20°C and the reaction mixture was continued to stir at 50°C for "25 30 minutes. Then, the resulting mixture was distilled under reduced pressure to remove the excess thionyl chlorice completely. Dioxane (25 ml) was added to the m 100 1 resulting acid-chloride and cycloartenol (5.0 g, 0.012 mole) dissolved in pyridine (25 ml) was added thereto at 0°C. The mixture was stirred at 60 0 C for minutes, then the resulting mixture was concentrated under reduced pressure, the residue was dissolved in chloroform (50 ml), and the solution was washed with saturated aqueous solution of Sodium bicarbonate. The aqueous layer was extracted with chloroform (3x50 ml).

The combined extracts were dried and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (solvent: chloroformhexane giving cycloartenyl-p-nitrobenzoate (6.3 g) in a 93 yield. m.p. 221 222 0

C.

22.5 Specific rotation [a]225 +62.30 (C 1.01, CHCl 3 Analysis, Calcd. for C 3 7

H

5 3 04N 575.80): C, 77.17; H, 9.28; N, 2.43. Found: C, 77.25; H, 9.21; N, 2.50.

-1 IR), KBr (cm 2930, 2850, 1715, 1520, 1350, 1290, 1120, 1100.

PMR (CDC13)6: 0.41 (1H, 1/2 ABq, 4.2 Hz), 0.61 (1H, 1/2 ABq, 4.2 Hz), 0.70-2.40 (27H, m), 0.91 (3H, 0.98 (3H, 1.05 (3H, s), 1.60 (3H, bs), 1.68 (3H, bs), 4.60-5.30 (2H,m), 8.00-8.50 (4H, m).

S' 25 Example 19 Preparation of cycloartenyl-p-aminobenzoate Cycloartenyl-p-nitrobenzoate (5.0 g, 8.70 mmoles) i 101 1 prepared according to the procedure of Example 18 was suspended in acetic acid (250 ml). By adding zinc powder (10.0 the mixture was stirred at 20 25 0

C

for 2.5 hours. Then, zinc powder was removed by filtration and the filtrate was concentrated under reduced pressure. The residual solid was dissolved in chloroform, and the solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with chloroform. The combined extracts were dried, and distilled to remove the solvent. The resulting residue was purified by silica gel column chromatography (solvent: methylene 't chloride), giving cycloartenyl-p-aminobenzoate (4.0 g) in a 84 yield. m.p. 168 169 0

C.

25 15 Specific rotation [D +62.20 (C 1.00, CHC1 3 Analysis, Calcd. for C 37

H

55 0 2 N 545.82): C, 81.41; H, 10.16, N,.2.57, Found: C, 81.52; H, 10.12; N, 2.53.

IRV, KBr 3470, 3350, 2930, 2850, 1705, 20 1685, 1625, 1600, 1515, 1310, 1275, 1170, 1115.

PMR (CDC1 3 0.36 (1H, 1/2 ABq, 4 Hz), 0.58 (1H, 1/2 ABq, 4 Hz), 0.70-2.30 (27H, 0.95 (3H, 0.98 (6H, 1.00 (3H, 1.59 (3H, bs), t 1.66 (3H, bs), 4.50 (2H, bs), 4.50-5.30 (2H, m), St C S25 6.40-6.80 (2H, 7.60-8.10 (2H, m).

Example Preparation of cycloartenyl-p-acetamidobenzoate 102 1 Dioxane (110 ml) was added to p-acetamidobenzoic acid (5.5 g, 0.031 mole). Thionyl chloride (21.0 ml, equivalents) and pyridine (0.5 ml) were added dropwise to the former solution at 20 0 C and the reaction mixture was stirred at 50°C for 5 minutes. The resulting mixture was concentrated under reduced pressure to remove the unreacted thionyl chloride. Dioxane (50 ml) and a solution of cycloartenol (10.0 g, 0.023 mole) in benzene (50 ml) were added to the concentrate and then pyridine (20 ml) was added at 20 0 C. This reaction mixture was heated at 70 0 C for 3 hours. Then the solvent was removed by distillation under reduced r' t €pressure the resulting residue was dissolved in chloroform (100 ml), and the solution was washed with tittle c c/ 15 saturated aqueous solution of sodium bicarbonate.

C r t C r tt The aqueous layer was extracted with chloroform (5x100 ml). The combined extracts were dried, concentrated under reduced pressure and the residue was purified by t ft silica gel column chromatography (solvent: chloroform- 20 ethyl acetate, giving cycloartenyl-pacetamidobenzoate (11.0 g) in a 80 yield. m.p. 202 204 C.

24 Specific rotation (a]D +59.60 (C 0.99, CHC13) Analysis, Calcd. for C 39

H

57 0 3 N 587.85): 25 C, 79.68; H, 9.77; N, 2.38. Found: C, 79.59; H, 9.82; N, 2.34.

IRV, ,Br (cm 3310, 2930, 2850, 1705, 1680, h.

h L i 1 103 1 1598, 1520, 1310, 1285, 1260, 1180, 1135.

PMR (CDC1 3 0.39 (1H, 1/2 ABq, 4 Hz), 0.60 (1H, 1/2 ABq, 4 Hz), 0.6C-2.30 (27H, 0.91 (6H, 0.98 (3H, 1.03 (3H, 1.60 (3H, bs), 1.68 (3H, bs), 2.20 (3H, 4.50-5.30 (2H, m), 7.51 (1H, bs), 7.59 (2H, 1/2 ABq, 8.4 Hz), 7.97 (2H, 1/2 ABq, 8.4 Hz).

Example 21 Preparation of cycloartenyl-p-aminobenzoate Tetrahydrofuran (200 ml) and 30 hydrochloric acid (100 ml) were added to cycloartenyl-p-acetamidobenzoate (10.0 g, 0.017 mole) prepared according to the procedure of Example 20. The mixture was refluxed Sfor 2 hours. Then the solvent was removed by distillation under reduced pressure. The residue was dissolved in chloroform (300 ml), and washed with 1N aqueous sodium hydroxide (200 ml) followed by saturated brine.

The aqueous layer and brine were extracted with chloroform (3x200 ml). The combined extracts were dried and 20 concentrated under reduced pressure, and the residue was purified by silica gel column chromatography t (solvent: ethyl acetate-hexane, giving cycloartenyl-p-aminobenzoate (3.2 g) in a 34 yield.

m.p. 168 169 0

C.

S* c( 25 Specific rotation [a]D +62.20 Analysis, Calcd. for C 37

H

55 0 2 N 545.82): C, 81.41; H, 10.16; N, 2.57. Found: C, 81.48; 4' 104 1 H, 10.23; N, 2.54.

Example 22 Preparation of cycloartenyl-m-nitrobenzoate Cycloartenol (15.0 g, 0.035 mole) was dissolved in pyridine (150 ml). m-Nitrobenzoyl chloride (8.5 g, 1.3 equivalents) was added dropwise to the solution at 0 C and the mixture was stirred at 40 0 C for 30 minutes.

Then, the resulting mixture was concentrated under reduced pressure and the residue was dissolved in methylene chloride (100 ml). The solution was washed with saturated aqueous solution of sodium bicarbonate.

The aqueous layer was extracted with methylene chloride **(3x100 ml). The combined extracts were dried and o concentrated under reduced pressure. The concentrate S 0 0 0 15 was dissolved in methylene chloride (50 ml), and f 0 crystals separated out by adding methanol (100 ml), *0*O giving cycloartenyl-m-nitrobenzoate (19.5 g) in a 96 yield. m.p. 162.5 163.5 0

C.

22 o Specific rotation [a]22 +60.80 (C 0.99, CHC1 3 D *3 20 Analysis, Calcd. for C37H5304N 575.80): 0« 37 53 4 C 77.17; H, 9.28; N, 2.43. Found: C, 77.10; H, 9.33; N, 2.37.

S-1 i IR), KBr (cm-1) 2920, 2850, 1712, 1532, 1350, 1290, 1145, 980, 715.

25 PMR (CDCl 3 0.42 (1H, 1/2 ABq, 4.2 Hz), 0.64 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.40 (27H, m), 0.92 (6H, 0.98 (3H, 1.07 (3H, s),

J

105 1 1.60 (3H, bs), 1.67 (3H, bs), 4.60-5.30 (2H, m), 7.50-8.50 (4H, m) Example 23 Preparation of cycloartenyl-m-aminobenzoate Cycloartenyl-m-nitrobenzoate (15.0 g, 0.026 mole) prepared according to procedure of Example 22 was suspended in acetic acid (750 ml). By adding zinc powder (30.0 the mixture was stirred at 40 0 C for 2 hours.

Then, stirred the zinc powder was removed by filtration and the filtrate was concentrated under reduced pressure.

The residue was dissolved in chloroform, and the solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with chloroform (5x300 ml). The combined extracts 15 were dried and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (solvent: chloroform), giving cycloartenyl-m-aminobenzoate (12.6 g) in a 89 yield.

r, rm.p. 172.5-173.5 0

C.

St 25.5 20 Specific rotation [a]D +62.60 (C 0.99, CHC1 3 Analysis. Calcd. for C 37

H

55 0 2 N 545.82): C 81.41; H 10.16; N 2.57. Found: C 81.53; H 10.11; N 2.53.

IR), KBr (cm 3450, 3350, 2900, 2850, 1700, 1627, 1460, 1287, 1240, 1100, 975, 755.

PMR (CDC1 3 0.40 (1H, 1/2 ABq, 4.2 Hz), 0.61 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.30 (27H, m), .4 1 0.91 (6H, 0.98 (3H, 1.03 (3H, s), 1.61 (3H, bs), 1.68 (3H, bs) 3.70 (2H, bs), 4.60-5.30 (2H, m) 6.70-7.50 (4H, m).

Example 24 Preparation of cycloartenyl nicotinate Toluene (10 ml) was thionyl chloride (50 ml, equivalents) were added to nicotinic acid (8.7 g, 0.071 mole) at 0°C, and the whole was stirred at for 30 minutes. The reaction mixture was concentrated under reduced pressure, and cycloartenol (20.0 g, 0.047 mole), pyridine (100 ml) and toluene (50 ml) were added thereto at 0°C, and the reaction mixture S* e0 was stirred at 100°C for 30 minutes. Then, the solvent r a was removed by distillation under reduced pressure, S 15 the resulting residue was dissolved in methylene chloride and the solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with methylene (3x300 ml). The combined extracts was concentrated to dryness under reduced pressure, and the crude crystals were recrystallized from methylene chloride-methanol (1:4, giving cycloartenyl-nicotinate (24.4 g) in a 98 yield, m.p. 170.5 1710C.

St 25 Specific rotation [a] 5 +67.80 (C 1.00, CHC1 3 Analysis, Calcd. for C 36

H

53 0 2 N 531.79): C 81.30; H 10.05; N 2.63. Found: C 81.22, H 10.09, N 2.57.

107 1 IRv, KBr 2920, 2850, 1718, 1590, 1282, 1122, 965, 740.

PMR (CDCl 3 0.40 (1H, 1/2 ABq, 4.2 Hz), 0.62 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.40 (27H, m), 0.91 (6H, 0.98 (3H, 1.04 (3H, s), 1.60 (3H, bs), 1.67 (3H, bs), 4.50-5.30 (2H, m), 7.20-9.30 (4H, m).

Example Preparation of cycloartenyl linolate Benzene (61 ml) was added to linoleic acid (23.0 g, 0.082 mole) and the solution was stirred at 0°C. Thionyl chloride (61 ml, 10 equivalents) was added dropwise thereto, and the reaction mixture was stirred at 40 0 C for 30 minutes. Then the resulting mixture was concentrated under reduced pressure, and benzene (125 ml) was added with stirring at 0°C.

Cycloartenol (25.0 g, 0.059 ml) dissolved in pyridine (125 ml) was added thereto and the whole was stirred S" at 40 0 C for 10 minutes. Then the solvent was removed 20 by distillation under reduced pressure. The resulting ,residue was dissolved in methylene chloride, and the solution was washed with saturated aqueous solution of Ssodium bicarbonate, and the aqueous layer was extracted with methylene chloride (3x500 ml). The combined extracts were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting oily material was purified by silica gel column 108 1 chromatography (solvent: methylene chloride-hexane giving cycloartenyl linolate (36.1 g) in a 90 yield. m.p. (oily matter).

21.5 Specific rotation +39.30 Analysis, Calcd. for C48H8002 689.12): C 83.65; H 11.70. Found: C 83.52; H 11.83.

-1 IRv, neat (cm- 2910, 2850, 1730, 1460, 1373, 1170, 980.

PMR (CDC13)6: 0.33 (1H, 1/2 ABq, 4.2 Hz), 0.56 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.50 (52H, m), 0.84 (3H, 0.87 (6H, 0.96 (3H, s), 1.60 (3H, bs) 1.67 (3H, bs) 2.76 (2H, bt, t 5.0 Hz), 4.40 4.80 (1H, 4.90-5.64 (5H, m) Example 26 Preparation of cycloartenyl-3,4-diacetoxybenzoate Toluene (50 ml) was added to 3,4-diacetoxybenzoic acid (20.9 g, 0.088 mole) and stirred at 0°C.

Thionyl chloride (33 ml, 5 equivalents) was added drocwise to 3,4-diacetoxybenzoic acid (20.9 g, 0.088 20 mole) and the mixture was stirred at 70° for 10 minutes.

t 6l SThe resulting mixture was concentrated under reduced pressure. Toluene (125 ml) was added to the concentrate at 0°C. Cycloartenol (25.0 g, 0.059 mole) j *dissolved in pyridine (60 ml) was added thereto and the mixture was stirred at 22'C for 1 hour. Then the solvent was removed by distillation under reduced pressure. The resulting residue was dissolved in 109 methylene chloride, and the solution was washed with saturated aqueous solution of sodium bicarbonate.

The aqueous layer was extracted with methylene chloride (3x400 ml). The combined extracts were dried and concentrated to dryness under reduced pressure. The residue was recrystallized from chloroform-methanol V/V) giving cycloartenyJ.-3,4-diacetoxybenzoate (33.8 g) in a 89 yield. m.p. 148 148.5 0

C

23 Specific rotation [a]23 +53.3 (C 0.99, CHC1 3 Analysis, Calcd. for C 4 1

H

58 0 6 646.87): C 76.12, H 9.04. Found: C76.01, H 9.17.

-1 IRy, KBr (cm 1 2910, 2850, 1775, 1710, 1610, M 1498, 1420, 1370, 1280, 1195, 1160.

PMR (CDCI 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.61 15 (1H, 1/2 ABq, 4.2 Hz), 0.70-2.40 (27H, m), ~0.90 (6H, 0.97 (3H, 1.02 (3H, s), 1.60 (3H, 1.68 (3H, bs), 2.29 (6H, s), 4.60-5.30 (2H, 7.10-7.50 (2H, 7.78o" 8.20 (3H, m).

20 Example 27 Preparation of cycloartenyl-3,4-dihydroxybenzoate Dioxane (460 ml) was added to cycloartenyl- S3,4-diacetoxybenzoate (23.0 g, 0.036 mole) prepared according to procedure of Example 26 and the solution was cooled to 0°C. After addition of 25 aqueous ammonia (46 ml) dropwise thereto, the mixture was stirred at 200C for 30 minutes and at 400C for

C

110 1 minutes to complete the reaction. Then the solvent was removed by distillation under reduced pressure.

The resulting residue was recrystallized from chloroform-hexane giving cycloartenyl-3,4dihydroxybenzoate (18.6 g) in a 93 yield. m.p.

199 199.5 0

C.

Specific rotation [aj2 6 +63.70 (C 0.97, CHC1 3 Analysis, Calcd. for C37H5404 562.80): C 78.96; H 9.67. Found: C 78.82; H 9.63.

-1 IRv, KBr (cm- 1 3470, 3330, 2900, 2850, 1705, 1679, 1605, 1525, 1435, 1280, 1230, 1098, 975.

PMR (CDC1 3 0.38 (1H, 1/2ABq, 4.2Hz), 0.50 (1H, 1/2ABq, 4.2Hz), 0.50-2.40 (27H, 0.88 (6H, 0.96 (3H, 1.00 (3H, 1.60 (3H, bs), 1.67 (3H, bs), 15 4.50-5.30 (2H, 5.80-6.80 (1H, bs), 6.70-7.90 (3H, m).

Example 28 Preparation of cycloartenyl-4-acetoxy-3-methoxybenzoate ,Toluene (26 ml) and thionyl chloride (23 ml) S 20 were added to 4-acetoxy-3-methoxybenzoic acid (13.0 g, 0.062 mole). The reaction mixture was stirred at 0 C for 20 minutes. Then the resulting mixture was concentrated to dryness under reduced pressure. To the residue were added pyridine (25 ml) and toluene (25 ml), and the mixture was stirred at 0°C. Cycloartenol (20.0 g, 0.047 mole) dissolved in pyridine ml) was added dropwise thereto, and the mixture 1i.

t' *r 1 was heated with stirring at 600C for 1.5 hours, at 800C for 1 hour, and at 1000C for 2 hour to complete the reaction. The resulting mixture was concentrated under reduced pressure, and the residue was dissolved in chloroform (300 ml). The solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with chloroform (3x300 ml).

The combined extracts was dried and concentrated under reduced pressure and the residue was purified by silica gel column chromatography (solvent: ethyl acetatehexane, giving cycloartenyl-4-acetoxy- 3-methoxybenzoate (26.0 g) in a 90 yield. m.p.

157 158 0

C.

23 Specific roatation [a]D +58.90 (C 0.99, CHC13 Analysis, Calcd. for C 4 0

H

5 8 0 5 618.86): C 77.63; H 9.45. Found: C 77.75; H 9.40.

-1 IRv, KBr (cm- 2920, 2850, 1770, 1710, 1600, 1500, 1410, 1280, 1210, 1195, 1170, 1100, 1030.

PMR (CDCl 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.60 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.40 (27H, m), 0.89 (6H, s) 0.96 (3H, 1.02 (3H, s), 1.60 (3H, bs), 1.68 (3H, bs), 2.32 (3H, s), 3.86 (3H, 4.60-5.30 (2H, 6.80-7.90 (3H, m).

Example 29 Preparation of cycloartenyl-4-hydroxy-3-methoxybenzoate 11I 1 Dioxane (480 ml) was added to cycloartenyl- 4 -acetoxy-3-methoxybenzoate (24.0 g, 0.039 mole) prepared according to the procedure of Example 28.

Thereto was added dropwise 25 aqueous ammonia (48 ml) at 0°C and the whole was stirred at 35 0 C for 1 hour.

Then the resulting mixture was concentrated under reduced pressure and the residue was dissolved in chloroform (200 ml), and the solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with chloroform (3x200 ml).

The combined extracts was dried and concentrated, and crystallized by adding ethanol, giving cycloartenyl- I t 4-hydroxy-3-methoxybenzoate containing a molecular E 4 t. ethanol (21.0 g) in a 87 yield. m.p. 132 133 0

C.

26 15 Specific rotation +61.30 (C 1.00, CH C1) cD I" Analysis, Calcd. for C 3 8

H

5 6 04-C 2

H

5 OH 622.90): C 77.12; H 10.03. Found: C 77.21; H 10.12.

-1 IRV, KBr 3380, 2920, 2850, 1705, 1683, 1607, 1590, 1510, 1280, 1225.

c C S 20 PMR (CDCl 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.60 t C (1H, 1/2 ABq, 4.2 Hz), 0.70--2.40 (27H, m), 0.91 (6H, 0.97 (3H, 1.03 (3H, s), S~ 1.60 (3H, bs), 1.68 (3H, bs), 3.93 (3H, s), 4.60-5.30 (2H, 6.08 (1H, bs), 6.78-7.80 (3H, m).

Example Preparation of cyclobranyl-3,4-diacetoxycinnamate n .si o .08 mole) 1 Thionyl chloride (30 ml) was added to a suspension of 3,4-diacetylcaffeic acid (21.44 g, 0.081 mole) in toluene (100 ml). The mixture was stirred at for 3 hours. The resulting mixture was distilled under reduced pressure to remove the solvent. The residue was suspended in a mixture of toluene (150 ml) and pyridine (30 ml). Cyclobranol (25 g, 0.0567 mole) was added to the suspension, and the mixture was stirred at 60 0 C for 2 hours. Then the solvents were removed by distillation under reduced pressure. The residue was extracted with chloroform (300 ml). The extract was washed, dried, and evaporated to remove the chloroform. The residue was purified by silica t gel column chromatography (solvent: toluene), giving cyclobranyl-3,4-diacetoxycinnamate (23.7 g) in a 61 yield. m.p. 174 175 0

C.

Specific rotation [a21 5 +37.00 (C 1.00, CHCl 3 Analysis. Calcd. for C44H6206 686.94): C 76.93; H 9.10. Found: C 76.87; H 9.15.

IRy, KBr 3400, 2850, 1775, 1705, 1200, tt** 1170.

SPMR (CDC1 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.62 1 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.50 (27H, m), 0.88 (6H, 0.96 (6H, 1.62 (9H, s), 1.28 (3H, 4.50-4.88 (1H, 6.39 (1H, 1/2 ABq, 15.6 Hz), 7.08-7.43 (3H, 7.62 (1H, 1/2 ABq, 15.6 Hz).

Thionyl chloride (17.43 ml, 2 equivalents) was added to a suspension of 3,4-dipropionylcaffeic acid (35.07 g, 0.12 mole) in toluene (150 ml), and the reaction mixture was stirred at 60 0 C for 2 hours. The resulting mixture was distilled under reduced pressure to remove the solvent. The residue was suspended in a mixture of toluene (240 ml) and pyridine (50 ml).

Cyclobranol (40 g, 0.0908 mole) was added to the suspension, and the mixture was stirred at 60°C for 2 hours. Then the solvents were removed by distillation under reduced pressure. The residue was extracted i with chloroform (300 ml). The extract was dried and it evaporated to remove the chloroform. The resulting residue was purified by silica gel column chromatography (solvent: toluene), giving cyclobranyl-3,4dipropionyloxycinnamate (41.9 g) in a 64 yield.

m.p. 163 165 0

C.

21.5 Specific rotation [a]D +34.70 (C 1.00, CHC13) 8, 0 Analysis, Calcd. for C 46

H

66 0 6 714.99): C 77.27; H 9.30. Found: C 77.34; H 9.23.

*i IRV, KBr (cm 2920, 2850, 1770, 1710, 1250, 1170.

PMR (CDC1 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.62 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.20 (27H, m), 0.90 (6H, 0.96 (6H, 1.25 (6H, t, 7.2Hz), 115 1 1.62 (9H, 2.57 (4H, q, 7.2 Hz), 4.50-4.88 (1H, 6.36 (1H, 1/2ABq, 15.6Hz), 7.05 (3H, 7.60 (1H, 1/2 ABq, 15.6 Hz).

Example 32 Preparation of cyclobranyl-3,4-dihydroxycinnamate Cyclobranyl-3,4-dipropionyloxycinnamate (35 g, 0.049 mole) prepared according to the procedure of Example 31 was dissolved in dioxane (600 ml). To the solution was added 25 aqueous ammonia (70 ml) and the mixture was stirred at 50 0 C for 2 hours. Then the solvent was removed by distillation under reduced pressure. The crude crystals were recrystallized from acetone-water giving cyclobranyl-3,4dihydroxycinnamate (22.3 g) in a 75 yield.

m.p. 246 247.5 0

C.

Specific rotation [ac] +33.60 (C 1.10, CHC13) Analysis, Calcd. for C 40

H

58 0 4 602.86): C 79.69; H 9.70. Found: C 79.62, H 9.68.

-1 ~IRV, KBr (cm1) 3400, 2920, 2850, 1680, 1600, 1520, 1440, 1275, 118C, 970.

S. PMR (CDC1 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.59 (1H, 1/2 ABq, 4.8 Hz), 0.69-2.20 (27H, m), S* 0.85 (3H, 0.90 (3H, s) 0.96 (6H, s), 1.60 (9H, 4.36-4.80 (1H, 6.16 (1H, 1/2 ABq, 15.6 Hz), 6.60-7.16 (5H, 7.74 (1H, 1/2 ABq, 15.6 Hz).

i

"I'

11i 4 4 4 4

S

144o*4 II 4$ 4 44 *i t a.

14 4 4 SExample 33 Preparation of cyclobranyl-3,4-dihydroxycinnamate The title compound was prepared according to following the procedure of Example 32 where cyclobranyl-3,4-diacetoxycinnamate (35 g, 0.051 mole) was used in place of cyclobranyl-3,4-dipropionyloxycinnamate. The yield was 22.1 g (64 m.p. 246 247 0

C.

25 Specific rotation [a]D +33.60 (C 0.11, CHCl 3 Analysis, Calcd. for C 40

H

58 0 4 602.86): C79.69; H 9.70. Found: C 77.78; H 9.62.

Example 34 Preparation of cyclobranyl-p-acetoxycinnamate p-Acetyl coumaric acid (18.2 g, 0.0885 mole) suspended in toluene (100 ml) and thionyl chloride (12.82 ml, 2 equivalents) was heated at 60 0 C for 2 hours with stirring. The resulting mixture was distilled under reduced pressure to remove the solvent. The residue was dissolved in a mixture of 20 toluene (150 ml) and pyridine (30 ml), and to the mixture was added cyclobranol (30 g, 0.068 mole).

The whole was heated at 60 0 C for 2 hours with stirring.

Then the solvents were removed by distillation under reduced pressure, and the residue was extracted with chloroform (250 ml). The extract was dried and evaporated to remove the chloroform. The residue was purified by silica gel column chromatography (solvent: 1 toluene) giving cyclobranyl-p-acetoxycinnamate (32.1 g) in a 75 yield. m.p. 164 165'C.

Specific rotation [a] 21 5 +40.80 (C 1.00, CHCl 3 Analysis, Calcd. for C 42

H

60 0 4 628.90): C 80.21; H 9.62.. Found: C 80.14; HI 9.67.

IRv, KBr (cm )2920, 2850, 1765, 1700, 1630, U 1370, 1200, 1165.

PMR (CDCl 3 0 .35 (1H, 1/2 ABq, 4 .8 HZ) 0 (1H, 1/2 ABg, 4.8 Hz), 0.70-2.40 (27H, m), 0 .90 (6H, 0 .98 (6H s) 1 .6 2 (9H, s), 2.28 O3H, s) 4.50-4.88 (1H, mn), 6.37 (11i, 1/2 ABq, 15.6 Hz) 6 .90-7 .20 (2H, 7.35tI 7.64 (2H, mn), 7.62 (1H, 1/2 ABg, 15.6 Hz).

Example_ Preparation of cyclobranyl-p-hydroxcinnamate Cyclobranyl-p-acetxycihnria.te (27 g, 0.043 mole) prepared according to the procedure of Example 34 was dissolved in Letrhdrofuran (400 ml) and Iheated with 25 aqt'eoas, vrmonia (50 ml) at 40-C for 2 hours with stL~ring. The rt~su2 ting mixture was evaporated to dryness under reduced pressure to remove the solvent. The crystals separating out were recrystallized from acetone-water giving cyclobranyl-p-hydroxycinnamate (22.2 g) in a 88 yield, m.p. 243 244*C.

Specific rotation (a 5+41.30 (C 1.09, CHC1 3 Analysis, Calcd. for C 40

H

58 0 3 586.86): ~LULilllll III~LLU l C I1CIC- CI L ;;3i P* 118 1 C 81.86; H 9.96. Found: C 81.77; H 9.99.

-1 IRv, KBr (cm 1 3370, 2920, 2850, 1670, 1605, 1585, 1510, 1280, 1170, 830.

PMR (CDC1 3 0.37 (1H, 1/2 ABq, 4.8 Hz), 0.62 (1H, 1/2 ABq, 4.8 Hz), 0.68-2.15 (27H, m), 0.88 (3H, 0.90 (3H, s) 0.97 (6H, s), 1.59 (9H, 4.39-4.78 (1H, 6.11 (1H, 1/2 ABq, 15.6 Hz), 6.55-6.83 (2H, 7.10- 7.41 (3H, 7.42 (1H, 1/2 ABq, 15.6 Hz).

Example 36 Preparation of cyclobranyl-4-acetoxy-3-methoxybenzoate To 4-acetylvanillic acid (18.60 g, 0.0885 mole) t dissolved in toluene (100 ml) was added thionyl chloride (12.86 ml, 2 equivalents) and the mixture was stirred at 60 0 C for 2 hours. The resulting mixture was evaporated under reduced pressure and the residue was dissolved in toluene (150 ml) and i I i pyridine (30 ml). To the solution was added cyclobranol (30 g, 0.068 mole) and the mixture was stirred at 60 0 C for 2 hours. Then the mixture was evaporated under the reduced pressure, and the residue was extracted with chloroform (300 ml). The extract was dried, concentrated in vacuo, and purified by silica gel column chromatography (solvent: 'thylacetatehexane, giving cyclobran)l-4-acetoxy-3methoxybenzoate ester (33.38g) in a 77 yield.

F- -aorsr-Pa~- -u_ 11) S m.p. 177 178 0

C.

21.5 Specific rotation [a]D +54.10 (C 1.02, CHC13) An&alysis, Calcd. for C 4 1 H6005 632.89): C 77.80; H 9.56. Found: C 77.71; H 9.64.

IRv, KBr (cm 2920, 2850, 1770, 1710, 1285, 1190, 1170.

PMR (CDCl 3 0.38 (1H, 1/2 ABq, 4.8 Hz), 0.63 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.40 (27H, m), 0.92 (6H, s) 0.98 (3H, s) 1.04 (3H, s), 1.62 (9H, 2.32 (3H, 3.88 (3H, s), 4.60-5.00 (1H, m) 6.92-7.20 (1H, m) 7.51- 7.80 (2H, m).

Example 37 Preparation of cyclobranyl-4-hydroxy-3-methoxy- 15 benzoate ':4 Cyclobranyl-4-acetoxy-3-methoxybenzoate (30 g, 0.0474 mole) prepared according to the procedure of Example 36 was dissolved in tetrahydrofuran (300 ml), and to" the solution was added 25 aqueous ammonia (60 ml). The mixture was stirred at 50 0 C for 2 hours, c then the resulting mixture was evaporated to dryness under reduced pressure. The residual crystals were recrystallized from acetone-water giving S' cyclobranyl-4-hydroxy-3-methoxybenzoate (23.1 in a 82 yield. m.p. 191 1930C.

26 Specific rotation [a]D +55.80 (C 1.02, CHC1 3 Analysis, Calcd. for C 39

H

58 0 4 590.85): i i 120 1 C 79.27; H 9.89. Found: C 79.35; H 9.80.

-1 IRy, KBr (cm 3400, 2920, 2850, 1700, 1590, 1510, 1275, 1220.

PMR (CDC13)6: 0.38 (1H, 1/2 ABq, 4.8 Hz), 0.64 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.2 (27H, m), 0.92 (6H, s) 1.0 (3H, s) 1.05 (3H, s), 1.64 (9H, 3.95 (3H, 4.6-5.0 (1H, m), 6.06 (1H, bs), 6.74-7.04 (1H, 7.44-7.77 (2H, s).

Example 38 Preparation of cyclobranyl-3,4-diacetoxybenzoate To diacetylprotocatechuic acid (21.08 g, 0.0885 mole) suspended in toluene (100 ml) was added thionyl S°chloride (12.86 ml, 2 equivalents), and the mixture o Swas stirred at 60 0 C for 2 hours. Then the resulting mixture was evaporated under reduced pressure. The 0 residue was suspended in toluene (150 ml) and pyridine ml), and to the suspension was added cyclobranol g, 0.068 mole). The mixture was stirred at 60 0

C

for 2 hours. Then the solution was evaporated under reduced pressure. The residue was extracted with chloroform (300 ml). And the extract was dried, concentrated in vacuo, and purified by silica gel Vat- column chromatography (solvent: toluene), giving S cyclobranyl-3,4-diacetoxybenzoate (24.2 g) in a 54 yield. m.p. 165 166 0

C.

Specific rotation [a 21.5 +51.3 (C 1.01, CHCl 3 Analysis, Calcd. for C 42

H

60 0 6 660.90): i i 12.1.

C 76.32; H 9.15. Found: C 76.45; H 9.10.

-1 IRy, KBr (cm 1 2920, 2850, 1770, 1715, 1280, 1195, 1160.

PMR (CDCl 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.62 (1H, 1'/2 ABq, 4.8 Hz), 0.70-2.40 (27H, m), 0.90 (6H, 0.96 (3H, 1.00 (3H, s), 1.60 (9H, 2.26 (6H, 4.60-4.98 (1H, m), 7.08-7.40 (1H, 7.73-8.06 (2H, m).

Example 39 Preparation of cyclobranyl-3,4-dihydroxybenzoate Cyclobranyl-3,4-diacetoxybenzoate (24.0 g, 0.0363 mole) prepared according to the procedure of Example 38 was dissolved in dioxane (480 ml) and to the solution was added 25 aqueous ammonia (48 ml).

The mixture was stirred at 40 0 C for 1 hour, then the resulting mixture was evaporated to dryness under reduced pressure. The residual crystals were recrystallized from ethanol, giving cyclobranyl-3,4dihydroxybenzoate (17.5 in a 84 yield. m.p.

215 216 0

C.

26 Specific rotation [a]D +59.40 (C 0.98, CHC1 3 Analysis, Calcd. for C 38

H

5 6 0 4 576.83): C 79.12; H 9.79. Found: C 79.03; H 9.87.

IRy, KBr (cm 3350, 2920, 2850, 1680, 1605, 1440, 1280, 1230, 1100, 975.

PMR (CDC13)S: 0.38 (1H, 1/2 ABq, 4.8 Hz), 0.61 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.20 (27H, m), 1 1i: i.

122 1 0.90 (6H, 0.98 (3H, 1.02 (3H, s), 1.62 (9H, 2.72-3.20 (2H, bs), 4.60-4.90 (1H, 6.70-6.96 (1H, 7.40-7.64 (2H, m).

Example Preparation of cyclobranyl-o-acetoxybenzoate To acetylsalicylic acid (15.94 g, 0.088 mole) suspended in toluene (100 ml) was added thionyl chloride (40 ml, 6.3 equivalents) and the mixture was stirred at 80 0 C for 3 hours. The resulting mixture was evaporated under reduced pressure, and the residue was dissolved in toluene (180 ml) and pyridine (40 ml), followed by addition of cyclobranol (30 g, 0.068 mole).

The mixture was continued to stir at 60 0 C for 2 hours, then the resulting mixture was evaporated under reduced pressure. The residue was extracted with chloroform (300 ml), and the extract was washed with 3 aqueous sodium bicarbonate, water, and saturated brine. The extract was dried, concentrated in vacuo, and the residue was purified by silica gel column chromatography (solvent: hexane-toluene, giving cyclobranyl-o-acetoxybenzoate (29.6 in a 72 S-yield. m.p. 165 166 0

C.

41 .23 i* Specific rotation [ajD +58.40 1.01, CHC13 I 1' Analysis, Calcd. for C40H5804 602.86): C 79.69; H 9.70. Found: C 79.64; H 9.78.

IRv, KBr (cm 1 2920, 2850, 1770, 1720, 1260, 1190, 1080 1 PMR (CDCl 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.60 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.40 (27H, m) 0.89 (6H, s) 0.96 (3H, s) 1.00 (3H, 1.62 (9H, 2.33 (3H, 4.60- 4.95 (1H, 6.88-7.68 (3H, m) 7.82- 8.10 (1H, m).

Example 41 Preparation of cyclobranyl-o-hydroxybenzoate Cyclobranyl-o-acetoxybenzoate (24 g, 0.0398 mole) prepared according to the procedure of Example was dissolved in dioxane (400 ml) and to the solution was added 25 aqueous ammonia (60 ml) dropwise. The mixture was stirred at 50 0 C for 2 hours and evaporated to dryness under reduced pressure. The residual crystals were recrystallized from acetone-water giving cyclobranyl-ohydroxybenzoate ester (20.2 in a 90 yield.

m.p. 200 201 0

C.

23 Specific rotation [a]D +69.30 (C 1.00, CHCl 3 Analysis, Calcd. for C 38

H

56 0 3 560.83): C 81.38; H 10.07. Found: C 81.29; H 10.02.

-1 A IR KBr (cm 3120, 2920, 2850, 1670, 1615, 1300, 1250, 1220, 1165, 1095.

PMR (CDCl 3 0.37 (1H, 1/2 ABq, 4.8 Hz), 0.62 (1H, 1/2 ABq, 4.8 Hz), 0.7-2.20 (27H, m), 1.05 (3H, 0.90 (6H, 0.97 (3H, s), 1.60 (9H, 4.60-5.00 (1H, 6.67-7.93 12,1 (4H, 10.92 (1H, s).

Example 42 Preparation of cyclobranyl-p-nitrobenzoate To p-nitrobenzoic acid (1 g, 0.006 mole) dissolved in dioxane (20 ml) was added thionyl chloride (3 ml, 7 equivalents) and the mixture was stirred at 0 C for 2 hours. The resulting mixture was evaporated under reduced pressure, then dioxane (20 ml) and pyridine (3 ml) were added to the resulting residue.

Cyclobranol (2 g, 0.0045 mole) was added to the solution and the mixture was stirred at 60 0 C for 2 hours to complete the reaction. The mixture was evaporated under reduced pressure, and the residue was extracted with chloroform (30 ml). The extract was washed successively with 3 aqueous sodium carbonate, water, and saturated brine, dried, and evaporated to dryness under reduced pressure. The crude crystals were washed with ethanol (50 ml), and recrystallized from ethyl acetate, giving cyclobranol-p-nitrobenzoate (2.28 g) in a 90 yield. m.p. 244 245 0

C.

24 Specific rotation +60.10 (C 1.00, CHC1 3 D 3 Analysis, Calcd. for C 3 8

H

5 5 0 4 N 589.83): C 77.37; H 9.40; N 2.37. Found: C 77.45; S' H 9.33; N 2.42.

IRv, KBr 2920, 2850, 1715, 1525, 1350, 1290, 1120, 1100, 720.

PMR (CDCl 3 0.39 (1H, 1/2 ABq, 4.8 Hz), 0.64

C

12b 1 (1H, 1/2 ABq, 4.8 Hz), 0.72-2.20 (27H, m), 0.92 (6H, 0.98 (3H, s) 1.06 (3H, s) 1.62 (9H, 4.59-5.02 (1H, 8.00-8.43 (4H, m).

Example 43 Preparation of cyclobranyl-p-aminobenzoate Cyclobranyl-p-nitrobenzoate (2 g, 0.0034 mole) prepared according to the procedure of Example 42 was suspended in acetic acid (80 ml), and was added zinc powder (2 g, 9 equivalents). The mixture was refluxed for 4 hours, then cooled and the zinc powder was O, separated by filtration. The filtrate was evaporated Sunder reduced pressure, and extracted with chloroform (50 ml). The extract was dried, concentrated in vacuo, a Do 15 and the purified by silica gel column chromatography (solvent: chloroform). The thus obtained crystals were recrystallized from ethanol, giving cyclobranylp-aminobenzoic acid ester (1.37 in a 72 yield.

o m.p. 190 191°C.

26 20 Specific rotation +58.40 (C 1.01, CHCl 3 93 Analysis, Calcd. for C 3 8

H

5 7 0 2 N 559.84): C 81.52; H 10.26; N 2.50. Found: C 81.49;

S

V H 10.21; N 2.57.

'o c -1 o* IRV, KBr (cm 3450, 3350, 2920, 2850, 1685, 1620, 1600, 1510, 1275, 1170, 1110.

PMR (CDC13)6: 0.37 (1H, 1/2 ABq, 4.8 Hz), 0.61 (1H, 1/2 ABq, 4.8 Hz), 0.72-2.30 (27H, m), i i i 12G 9980 .86 8 6t p 0*0 00 O0 p o 0&0 .8 "0406 pb 04* 4 0 00 6 .8 0 1 0.89 (6H, 0.96 (3H, 1.00 (3H, s), 1.60 (9H, s) 3.60-4.40 (2H, b) 4.50-4.90 (1H, 6.44-6.76 (2H, 7.64-8.00 (2H,m).

Example 44 Preparation of cyclobranyl-p- acetamidobenzoate To p-acetamidobenzoic acid (15.85 g, 0.088 mole) dissolved in dioxane (150 ml) was added thionyl chloride (25.7 ml, 4 equivalents) and the mixture was stirred at 600C for 2 hours. The resulting mixture was evaporated under reduced pressure and dioxane (150 ml) and pyridine (50 ml) were added to the residue. Cyclobranol (30 g, 0.068 mole) was added to the solution, and the mixture was stirred at 600C for 2 hours. The reactioh mixture was evaporated under reduced pressure, and to the residue was added ethyl acetate (300 ml). The crude crystals which separated out were filtered, and purified by silica gel column chromatography (solvent: chloroform). The resulting crystals were further recrystallized from ethanol, 20 giving cyclobranyl-p-acetamidobenzoate (32 in a 78 yield. m.p. 197 1980C.

24 Specific rotation[a]D 54.70 (C 1.01, CHC13) Analysis, Calcd. for C40H5903N 601.88): C 79.82; H 9.88; N 2.33. Found: C 79.75; H 9.83; N 2.41.

IRy, KBr 3420, 2920, 2850, 1710, 1690, 1680, 1600, 1535, 1280, 1175.

127 PMR (CDOD-CDC13)6: 0.38 (1H1, 1/2 ABq, 4.8 Hz), 0.62 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.28 (27H, m), 0.90 (6H, 0.98 (3H, 1.04 (3H, s), 1.62 (9H, 2.16 (3H, s) 4.50-4.96 (1H, m), 7.37 (1H, 7.44-8.12 (4H, m).

Example Preparation of cyclobranyl-p-aminobenzoate Cyclobranyl-p-acetamidobenzoate (32 g, 0.053 mole) prepared according to the procedure of Example 44 was dissolved in tetrahydrofuran (300 ml) and conc.

hydrochloric acid (60 ml) was added to it, and stirred at 70°C for 2 hours. Then the mixture was evaporated under reduced pressure. The residue was extracted with chloroform (400 ml), then the extract was dried and concentrated. The residue was purified twice by silica gel column chromatography (solvent: toluene-hexane-ethyl acetate, giving cyclobranyl-p-aminobenzoate (17 g) in a 57 yield.

m.p. 190 191 0

C.

Specific rotation [aj26 +58.40 (C 1.01, CHC1 3 Analysis, Calcd. for C 3 8

H

5 7 0 2 N 559.84): C 81.52; H 10.26; N 2.50. Found: C 81.57; H 10.34; N 2.44.

Example 46 Preparation of cyclobranyl-o-nitrobenzoate To o-nitrobenzoic acid (12.32 g, 0.074 mole) dissolved in dioxane (200 ml) was added thionyl rt 0 Q la~ 128 1 chloride (30 ml, 5.6 equivalents) and the mixture was stirred at 60 0 C for 2 hours. After the reaction was finished, the mixture was evaporated to dryness under reduced pressure. The resulting residue was dissolved in dioxane (150 ml) and pyridine (50 ml). To the solution was added cyclobranol (25 g, 0.0567 mole) and this mixture was stirred at 60 0 C for 2 hours.

Then the mixture was evaporated to dryness under reduced pressure, and the residue was extracted with chloroform (300 ml). The extract was washed successively with 3 aqueous sodium carbonate, water, and saturated brine, dried, and evaporated to dryness under 0 0 reduced pressure. The crude crystals were washed with o. ethanol (100 ml), and recrystallized from acetone- *0 15 water giving cyclobranyl-o-nitrobenzoate Po (30.8 in a 92 yield. m.p. 217 218 0

C.

Specific rotation [a] 2 1 +90.80 (C 0.98, CHC13 Analysis, Calcd. for C 38

H

55 0 4 N 589.83): C 77.37; H 9.40; N 2.37. Found: C 77.42, H 9.34; N 2.47.

1! cm -1 I. KBr (cm 2930, 2850, 1710, 1535, 1380, 1300.

PMR (CDC1 3 0.36 1/2 ABq, 4.8 Hz), 0.62 (1H, 1/2 ABq, 4.8 Hz), 0.75-2.20 (27H, m), 0.90 (3H, 0.92 (6H, 0.98 (3H, s), 1.64 (9H, s) 4.63-5.00 (1H, 7.46-7.98 (4H, m).

129 1 Example 47 Preparation of cyclobranyl-o-aminobenzoate Cyclobranyl-o-nitrobenzoic acid ester (30 g, 0.05 mole) prepared according to the procedure of Example 46 was suspended in acetic acid (1.2 And to the suspension was added zinc powder (30 g, 9 equivalents) which was washed with dilute hydrochloric acid before it. The mixture was refluxed for 5 hours, then cooled and the zinc powder was removed by filtration. The filtrate was evaporated under reduced pressure and the residue was extracted with chloroform a (300 ml). The extract was dried, concentrated to dryness in vacuo, and purified by silica gel column o"a chromatography (solvent: chloroform), giving cycloa a S° 15 branyl-o-aminobenzoate (20 g) in a 68 yield. m.p.

207 208 0

C.

Specific rotation []24 +74.9° (C 1.00, CHC1 3 Analysis, Calcd. for C38H5702N 559.84): e o C 81.52; H 10.26; N 2.50. Found: C 81.59; H 10.28; N 2.43.

IRV, KBr (cm-1) 60, 3350, 2940, 2850, 1670, 1620, 1290, 1245, 1155, 755.

l* PMR (CDC1 3 0.37 (1H, 1/2 ABq, 4.8 Hz), 0.62 r'o" 1 H 1/2 ABq, 4.8 Hz), 0.7-2.20 (27H, m), 0.92 (6H, 0.98 (3H, 1.04 (3H, s), 1.64 (9H, 4.50-4.95 (1H, 5.73 (2H, bs), 6.40-6.78 (2H, 7.00-7.40 (1H, m), I 130 7.61-7.98 (1H, m).

:-1 #4tS I CI r Cr C C 011 Example 48 Preparation of cyclobranyl-m-nitrobenzoate Cyclobranol (18 g, 0.04 mole) dissolved in S pyridine (200 ml) was stirred and cooled in a bath of ice water, and to it was added m-nitrobenzoyl chloride (9.85 g, 0.053 mole). Then the mixture was warmed to 0 C and continued to stir for 12 hours. After the reaction is complete the mixture was evaporated under reduced pressure. Ice cold water (200 ml) was added to the residue and crystals begans to appear. The crude crystals were separated by filtration and recrystallized from acetone-water giving cyclobranol-m-nitrobenzoate (20.1 in a 83 yield.

m.p. 206 207 0

C.

21 Specific rotation +60.80 (C 0.99, CHCl 3 Analysis, Calcd. for C 38

H

55 0 4 N 589.83): C 77.37; H 9.40; N 2.37. Found: C 77.44; H 9.33; N 2.45.

IRV, KBr 2930, 2850, 1715, 1530, 1350, 1290, 1140, 715.

PMR (CDC13)6: 0.40 (1H, 1/2 ABq, 4.8 Hz), 0.66 (1H, 1/2 ABq, 4.8 Hz), 0.80-2.20 (27H, m), 0.92 (6H, 0.98 (3H, 1.08 (3H, s), 1.64 (9H, 4.65-5.05 (1H, 7.44-7.80 (1H, 8.20-8.52 (2H, 8.72-8.89 (1H, m).

C

131 1 Example 49 Preparation of cyclobranyl-m-aminobenzoate Cyclobranyl-m-nitrobenzoate (24 g, 0.04 mole) prepared according to the procedure of Example 48 was suspended in acetic acid (1.3 Zinc powder .(24 g, 9 equivalents) was added to the suspension, and the mixture was refluxed for 2 hours. After the reaction was finished mixture was cooled and the zinc powder was separated by filtration. The filtrate was evaporated under reduced pressure, and the residue was extracted with chloroform (300 ml). The extract was dried, evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (solven t chloroform), giving cyclobranyl-m-aminobenzoate (19 g) in a 83 yield. m.p.

187 1880C.

24 Specific rotation [a]D +61.70 (C 1.03, CHC1 3 Analysis, Calcd. for C 38

H

57 0 2 N 559.84): C 81.52; H 10.26; N 2.50. Found: C 81.44; H 10.29; N 2.56.

Tc -1 C .4 IRv, KBr (cm 3450, 3350, 2940, 2850, 1700, 1460, 1320, 1290, 1245, 755.

S PMR (CDC13)6: 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.61 D (1H, 1/2 ABq, 4.8 Hz), 0.70-2.28 (27H, r 0.90 (6H, 0.98 (3H, 1.04 (3H, s), 1.64 (9H, s) 3.53 (2H, b) 4.59-4.90 (1H, m) 6.60-7.55 (4H, m) 132 *004 o tI *4 r a f2 I 41>4r 1 Example Preparation of cyclobranylnicotinate Thionyl chloride (30 ml, 4.6 equivalents) was added to nicotinic acid (10.9 g, 0.0885 mole) and the mixture was stirred at 80 0 C for 2 hours to complete the reaction. Then the excess thionyl chloride was removed under reduced pressure. To the residue dissolved in a mixture of toluene (120 ml) and pyridine (50 ml) was added cyclobranol (30 g, 0.068 mole) and the mixture was stirred at 60 0 C for 2 hours.

Then the mixture was evaporated to dryness under reduced pressure, and the residue was extracted with chloroform (300 ml). The extract was dried, concentrated in vacuo, and the residue was purified by silica 15 gel column chromatography (solvent: toluene), giving cyclobranylnicotinate (32.3 in a 87 yield.

m.p. 176 177 0

C.

26 Specific rotation a26 +63.40 (C 1.04, CHC1 3 Analysis, Calcd. for C37H5502N 545.82): C 81.41; H 10.16; N 2.57. Found: C 81.49; H 10.13; N 2.64.

IRV, KBr (cm 1 2920, 2850, 1720, 1590, 1285, 1120, 965.

PMR (CDC1 3 0.37 (1H, 1/2 ABq, 4.8 Hz), 0.62 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.27 (27H, m), 0.90 (6H, 0.93 (3H, 1.03 (3H, s), 1.62 (9H, 4.60-5.00 (1H, 7.13-7.50

I

r ic

C

CCC $2 1>440 4* 4 i 1 133 1 (1H, 8.06-8.40 (1H, 8.60-8.82 (1H, m), 9.07-9.27 (1H, m).

Example 51 Preparation of cyclobranyllinolate Thionyl chloride (20 ml, 4.8 equivalents) was added to linoleic acid (16.5 g, 0.0588 mole) and the mixture was stirred at 40 0 C for 1.5 hours. After the reaction was completed excess thionyl chloride was removed under reduced pressure, and the residue was dissolved in a mixture of toluene (100 ml) and pyridine ml). Cyclobranol (20 g, 0.0454 mole) was added to the solution and the mixture was stirred at 40 0 C for 0o tR 2 hours, then evaporated under reduced pressure. The oily residue was extracted with chloroform (300 ml), and the extract was dried and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (solvent: toluene-hexane, giving cyclobranyllinolate (22.2 in a 69 yield. ii.p. oily matter.

21.5 Specific rotation [a]D +36.50 (C 0.95, CHC1 3 Analysis, Calcd. for C 49

H

82 0 2 703.15): C 83.69; H 11.76. Found: C 83.61; H 11.88.

S^ IRv, KBr (cm) 2910, 2850, 1730, 1460, 1372, 1175.

PMR (CDC1 3 0.34 (1H, 1/2 ABq, 4.8 Hz), 0.58 (1H, 1/2 ABq, 4.8 Hz), 0.68-2.43 (52H, m), 0.83 (3H, 0.89 (6H, 0.95 (3H, s), 4< J 1 !i 134 0 0 0 48 0 0 00 00 44

CI

0f e C L 1 1.60 (9H, 2.52-2.91 (2H, 4.40-4.80 (1H, 5.04-5.62 (4H, m).

Example 52 Preparation of cyclobranyl-m-methoxybenzoate m-Anisic acid (8.1 g, 0.053 mole) was added to thionyl chloride (20 ml), and the mixture was stirred at 60°C for 2 hours. After the reaction is completed the excess thionyl chloride was removed under reduced pressure. The residue was dissolved in pyridine ml) and cyclobranol (18 g, 0.041 mole) was added to the solution. The mixture was stirred at 600C for 2 hours, then evaporated to dryness under reduced pressure. The resulting residue was purified by silica gel column chromatography, and recrystallized from ethanol, giving cyclobranyl-m-methoxybenzoate (20.7 g), in a 88 yield. m.p. 163 1640C.

21.5 Specific rotation [al 215 +61.6 (C 1.00, CHC1 3

D

Analysis, Calcd. for C39H 5 8 03 574.85): C 81.48; H10.17. Found: C 81.40; H 10.25.

IRy, KBr (cm- 1 2920, 2850, 1715, 1585, 1270, 1220, 1100, 750 PMR (CDC1 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.60 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.20 (27H. m), 0.90 (6H, 0.98 (3H, 1.02 (3H, s), 1.61 (9H, 3.82 (3H, 4.60-4.95 (1H, m), 6.84-7.72 (4H, m).

I),

133 1 Example 53 Preparation of cyclobranyl-m-acetoxybenzoate m-Acetoxybenzoic acid (19 g, 0.105 mole) and thionyl chloride (40 ml) was mixed and stirred at 60 0 C for 2 hours. After the reaction is complete, the excess thionyl chloride was removed by distillation, and the residue was dissolved in a mixture of toluene (220 ml) and pyridine (60 ml). Cyclobranol g, 0.0794 mole) was added to the solution and the mixture was stirred at 60 0 C for 2 hours. After that the solution was evaporated under reduced pressure, °oooo° and the residue was extracted with chloroform (300 ml).

94 The extract was dried, concentrated in vacuo, and the o I S residue was purified by silica gel column chromatott sa S 15 graphy (solvent: chloroform), giving cyclobranyl-macetoxybenzoate (46.3 in a 97 yield. m.p.

142 143 0

C.

21.5 SSpecific rotation [a]215 +59.00 (C 0.97, CHC1 3 Analysis, Calcd. for C 40H5804 602.86): C 79.69; H 9.70. Found: C 79.58; H 9.73.

IRv; KBr 2920, 2850, 1770, 1720, 1290, 1270, 1200, 1100.

PMR (CDCl 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.61 S(1H, 1/2 ABq, 4.8 Hz), 0.70-2.38 (27H, m), 0.90 (6H, 0.96 (3H, 1.02 (3H, s), 1.62 (9H, 2.31 (3H, 4.60-4.96 (1H, 7.11-8.02 (4H, m).

136 1 Example 54 Preparation of cyclobranyl-m-hydroxybenzoate Cyclobranyl-m-acetoxybenzoate (24.5 g, 0.0406 mole) prepared according to the procedure of Example 53 was dissolved in tetrahydrofuran (300 ml). To the solution was added 25 aqueous ammonia (40 ml) and the mixture was stirred at 40 0 C for 2 hours. The reaction mixture was evaporated to dryness under reduced pressure, and the resulting residue was recrystallized from ethanol, giving cyclobranyl-mhydroxybenzoate (20.3 in a 86 yield. m.p.

203 204.5 0

C.

21.5 2 Specific rotation [a21D +62.20 (C 0.95, CHCl 3 Analysis, Calcd. for C 3 8

H

5 6 0 4 576.83): o0 4 3 S15 C 79.12; H 9.79. Found: C 79.23; H 9.72.

06- IRv, KBr 3380, 2920, 2850, 1690, 1600, 1450, 1290, 1245, 1110, 760.

PMR (CDC1 3 0.38 (1H, 1/2 ABq, 4.8 Hz), 0.61 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.20 (27H, m), 0.93 (6H, 0.98 (3H, 1.04 (3H, s), 1.64 (9H, 3.54 (1H, bs), 4.58-4.90 (1H, 6.84-7.66 (4H, m).

P.

11 Example Sa Preparation of cyclobranyl-p-methoxybenzoate Cyclobranol (18 g, 0.041 mole) dissolved in pyridine (80 ml) was stirred and cooled in a bath of ice water, and to it was added p-methoxybenzoyl 13'? 1chloride (9.1 g, 0.0533 mole). The mixture was warmed to 20 0 C and continued to stir for 15 hours.

Then the mixture was evaporated under reduced pressure, and the residue was extracted with chloroform (300 ml).

The extract was concentrated in vacuo, and the resulting residue was purified by silica gel column chromatography (solvent. chloroform), giving cyclobranyl-pmethoxybenzoate (21.7 in a 92 yield. m.p. 202 203 0

C.

1 0 Specific rotation [al21.5 +60.20 (C 1.06, CHCl 3 Analysis, Calcd. for C39H5803 574.85): SO. C 81.48; H 10.17. Found: C 81.32; H 10.25.

00 IRV, KBr (cm 2920, 2850, 1705, 1605, 1510, o 1270, 1255, 1170, 1110, 1100.

o 15 PMR (CDC1 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.61 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.38 (27H, m), 0.90 (6H, 0.98 (3H, 1.02 (3H, s), 1.62 (9H, s) 3.84 (3H, s) 4.60-4.95 (1H, m), 6.72-7.04' (2H, 7.80-8.12 (2H, m).

Example 56 Preparation of cyclobranyl-o-methoxybenzoate Cyclobranol (18 g, 0.041 mole) dissolved in pyridine (80 ml) was stirred and cooled in a bath of c ice water, then to it was added o-methoxybenzoyl chloride (9 g, 0.0527 mole). The mixture was warmed to 20 0 C and continued to stir for 15 hours. The mixture was evaporated under reduced pressure, and il~ I- I 138 4P a a, O V~ 4 4404 4L the residue was extracted with chloroform (300 ml).

The extract was dried and the resulting residue was purified by silica gel column chromatography (solvent: chloroform), giving cyclobranyl-o-methoxybenzoate (21.9 in a 93 yield. m.p. 176 177 0

C.

21.5 Specific rotation [a]D +46.3 (C 1.00, CHC3) Analysis, Calcd. for C 39

H

58 0 3 574.85): C 81.48; H 10.17. Found: C 81.40; H 10.25.

-1 IRv, KBr (cm- 1 2920, 2850, 1695, 1600, 1460, 1300, 1250, 1130, 760.

PMR (CDC1 3 0.35 (1H, 1/2 ABq, 4.8 Hz), 0.59 (1H, 1/2 ABq, 4.8 Hz), 0.70-2.20 (27H, m), 0.90 (3H, 0.96 (6H, 0.99 (3H, s), 1.61 (9H, 3.86 (3H, 4.60-5.00 (1H, m), 6.72-7.94 (4H, m) Example 57 Preparation of cyclobranyl-p-acetoxybenzoate p-Acetoxybenzoic acid (18.6 g, 0.103 mole) and thionyl chloride (40 ml) were mixed and stirred at 60 0 C for 12 hours. Then, the excess thionyl chloride was removed by distillation under reduced pressure.

And to the residue suspended in toluene (220 ml) and pyridine (60 ml) was added cyclobranol (35 g, 0.0794 mole) and the mixture was stirred at 60°C for 2 hours.

Then the solution was evaporated under reduced pressure, and the residue was extracted with chloroform (400 ml). The extract was dried and concentrated V P 4 444 0 4. 44 4 40 04 a o 404 4 44,9 41 £444 4* QK9 *n A' 4 9 i 133 1 in vacuo, and the resulting product was purified by silica gel column chromatography (solvent: chloroform), giving cyclobranyl-p-acetoxybenzoate ester (44.1 g) in a 92 yield. m.p. 192 193 0

C.

22 Specific rotation [a]D +55.50 (C 0.93, CHC13) Analysis, Calcd. for C40H5804 602.86): C 79.69; H 9.70. Found: C 79.61; H 9.79.

-1 IRv, KBr (cm- 2920, 2850, 1765, 1715, 1270, 1190, 1160, 1115.

PMR (CDCl 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.60 (1H, 1/2 ABq, 4.8 Hz), 0.68-2.40 (27H, m), o 0.89 (6H, s) 0.96 (3H, 1.01 (3H, s), 1.60 (9H, 2.28 (3H, s) 4.60-5.00 (1H, m), 6.96-7.32 (2H, 7.90-8.25 (2H, m).

Example 58 Preparation of cyclobranyl-p-hydroxybenzoate Cyclobranyl-p-acetoxybenzoate (24 g, 0.0398 mole) prepared according to the procedure of Example 57 was dissolved in tetrahydrofuran (350 ml) and to that was added 25 aqueous ammonia (50 ml). The c mixture was continued to stir at 40 0 C for 1 hour, then, evaporated to dryness under reduced pressure.

'i The residual crystals were recrystallized from ethanol, I giving cyclobranyl-p-hydroxybenzoate (21.0 in a 94 yield. m.p. 174 175 0

C.

23 Specific rotation [cD 2 +57.70 (C 1.10, CHC1 3 Analysis, Calcd. for C 38

H

56 0 3

*C

2

H

5 OH (M.W.

xi 1410 1 606.9): C 79.16; H 10.30. Found: C 79.13; H 10.42.

IRV, KBr (cm 3400, 2920, 2850, 1685, 1610, 1280, 1160.

PMR (CDCl 3 0.38 (1H, 1/2 ABg, 4.8 Hz), .0.64 (1H, 1/2 ABq, 4.8 Hz) 0.70-2.42 (27H, n), 0.91 (6H, 0.98 (3H, 1.02 (3H, s), 1 .28 (3H, t, 7.2 Hz), 1.62 (9H, s) 3.77 (2H, q. 7.2 Hz), 4.60-5.00 (1H, in), 6.72-7.18 (4H, in), 7.80-8.12 (2H, mn).

riot.* Example 59 Preparation of cycloartenyl-4-acetoxy-3-ethoxya 8 benzoate The title compound was prepared according to following the procedure of Example 28 where 4-acetoxy- 3-ethoxybenzoic acid (13.9 g, 0.062 mole) was used in place of .4-acetoxy-3-methoxybenzoic acid (13.0g); I a' cycloartenyl-4--acetoxy-3-echoxybenzoate was yielded 24.5 g (82 m.p. 140 141'C.

Specific rotation [a]D +58.20. (C 1.00, CHCl 3 Analysis, Calcd. for C 41H 6 0 5(M.W. 632.89): C 77.80; H 9.56. Found: C H 9.43.

Example Preparation of cycloartenyl-3-ethoxy-4 -hydroxyben zoate Cycloartenyl-4-acetoxy-3-ethoxybenzoate (30 g, 0.047 mole) prepared according to the procedure of M MMM nrr*Pl~a*u 141 1 Example 59 was dissolved in tetrahydrofuran (300 ml), and to it was added 25 aqueous ammonia (60 ml).

After the mixture was stirred at 50 0 C for 2 hours, the mixture was evaporated to dryness under reduced pressure. The residual crystals were recrystallized from acetone-water giving cycloartenyl- 3-ethoxy-4-hydroxybenzoate ester (20.5 in a 74 yield, m.p. 128 130 0

C.

24 Specific rotation [a]D +59.5° (C 1.00, CHC 3 Analysis, Calcd. for C 39

H

58 0 4 590.85): C 79.27; H 9.89. Found: C 79.21; H 9.82.

Example 61 Preparation of cyclobranyl-4-acetoxy-3-ethoxybenzoate 0 The title compound was prepared according to 15 following the procedure of Example 28 where 4-acetoxy- 3-ethoxybenzoic acid (13.9 g, 0.062 mole) and cyclobranol (20.0 g, 0.045 mole) were used respectively, in place of 4-acetoxy-3-methoxybenzoic acid (13.0 g) and cycloartenol (20.0 cyclobranyl-4-acetoxy-3- 00 20 methoxybenzoate was yielded 23.4 g (80 m.p.

161 162 0

C.

24 Specific'rotation [a]D +56.50 (C 1.00, CHC13 Analysis, Calcd. for C 4 2

H

6 2 0 5 646.92): C 77.97; H 9.66. Found: C 78.05; H 9.61.

Example 62 Preparation of cyclobranyl-3-ethoxy-4-hydroxybenzoate Cyclobranyl-4-acetoxy-3-ethoxybenzoate (30 g, 142 1 0.046 mole) prepared according to procedure of Example 61 was dissolved in tetrahydrofuran (300 ml) and to it was added 25 aqueous ammonia (60 ml). After the mixture was stirred at 50 0 C for 3 hours, the mixture was evaporated to dryness under reduced pressure, and the residual crystals were recrystallized from acetonewater giving cycloartenyl-3-ethoxy-4hydroxybenzoate (20.8 in a 74 yield. m.p. 175 176 0

C.

24 Specific rotation [a]D +57.50 (C 1.00, CHC1 3 Analysis, Calcd. for C 40

H

60 0 4 604.88): C 79.42; H 10.00. Found: C 79.31; H 10,12.

Example 63 f t f Preparation of 24-methylenecycloartanyl-4-acetoxy- 15 3-ethoxybenzoate The title compound was prepared according to following the procedure of Example 28 where 4-acetoxy- 3-ethoxybenzoic acid (13.9 g, 0.062 mole) and 24methylenecycloartenol (20.0g, 0.045 mole) were used in place of 4-acetoxy-3-methoxybenzoic acid (13.0 g) and cycloartenol (20.0 respectively; The yield was 23.1 g (79 m.p. 152 153 0

C.

24 Specific rotation [a]D +57.8 (C 1.00, CHC1 3 Analysis, Calcd. for C42H620 646.92): C 77.97; H 9.66. Found: C 77.92; H 9.54.

Example 64 Preparation of 24-methylenecycloartanyl-3-ethoxy- 143 1 4-hydroxybenzoate The title compound-was prepared according to following the procedure of Example 55 where 24methylenecycloartanol-4-acetoxy-3-ethoxybenzoate (30 g, 0.046 mole) obtained by the procedure of Example 63 was used; The yield was 20.1 g (72 m.p. 141 1420C.

24 Specific rotation [a]D +58.40 (C 1.00, CHC1 3 Analysis, Calcd. for C40H6004 604.88): C 79.42; H 10.00. Found: C 79.49; H 10.05.

Example Preparation of cyclobranyl-4-acetoxy-3-methoxycinnamate To 4-acetoxy-3-methoxycinnamic acid (26.5 g, 15 0.112 mole) suspended in toluene (200 ml) was added thionyl chloride (16.3 ml, 3 equivalents) and the mixture was stirred at 60 0 C for 2 hours. Then the mixture was evaporated under reduced pressure and the residue was suspended in toluene (150 ml) and 20 pyridine (30 ml). Cyclobranol (33.1 g, 0.075 mole) was added to the suspension and the mixture was stirred at 60°C for 2 hours. After the reaction was complete, the mixture was evaporated to dryness under reduced pressure, and the residue was extracted with chloroform (300 ml). The extract'was washed, dried, and evaporated to dryness under reduced pressure.

The residual crystals were recrystallized from ethanol, (ft(

II

SI.

I'I

144 S giving cyclobranyl-4-acetoxy-3-methoxycinnamate (44.9 in a 91 yield. m.p. 175 176 0

C.

25 Specific rotation [a]D +37.00 (C 1.00, CHC1 3 Analysis, Calcd. for C 4 3

H

6 2 0 5 658.93): C 78.38; H 9.48. Found: 78.44; H 9.43.

Example 66 Preparation of cycloartenyl-4-acetoxy-3-methoxycinnamate The title compound was prepared according to following the procedure of Example 65 where cycloartenol (33 g, 0.077 mole) was used in place of cyclobranol (33 the yield was 42.4 g (86 m.p. 187 188 0

C.

24 Specific rotation [a24 +40.70 (C 1.00, CHCl 3 Analysis, Calcd. for C42H6005 644.90): C 78.22; H 9.38. Found: C78.34; H 9.30.

Example 67 Preparation of 24-methylenecycloartanyl-4-acetoxy- 3-methoxycinnamate The title compound was prepared according to following the procedure of Example 65 where 24methylenecycloartanol (33 g, 0.0749 mole) was used in place of cyclobranol; The yield was 43.7 g (88 m,p. 216 217 0

C.

24 Specific rotation [a]D +40.10 (C 1.00, CHC13J Analysis, Calcd. for C43H6205 658.93): C 78.38; H 9.48. Found: C 78.46; H 9.40.

;I

145 1 IRy, KBr (cm 2920, 2850, 1765, 1710, 1635, 1510, 1275, 1258, 1200, 1170, 1155.

PMR (CDCl 3 0.39 (1H, 1/2 ABq, 4.2 Hz), 0.61 (1H, 1/2 ABq, 4.2 Hz), 0.60-2.40 (28H, m), 0.82 (6H, 0.96 (6H, 1.02 (6H, d, 7.2 Hz), 2.32 (3H, 3.86 (3H, 4.70 (2H, bs), 4.50-4.90 (1H, 6.40 (1H, 1/2 ABq, 16 Hz), 6.90-7.30 (3H, 7.58 (1H, 1/2 ABq, 16 Hz).

Example 68 Preparation of cycloartenyl-4-acetoxy-3-ethoxycinnamate The title compound was prepared according to following the procedure of Example 65 where 4-acetoxy- 3-ethoxycinnamic acid (27.0 g, 0.108 mole) and cycloartenol (25.2 g, 0.059 mole) was used in place of 4acetoxy-3-methoxycinnamic acid (26.5 g) and cyclobranol (33 respectively; the yield was 32.6 g (83 m.p. 165 166 0

.C.

24 Specific rotation [a]D +40.50 (C 1.00, CHC13 Analysis, Calcd. for C 4 3

H

6 2 0 5 658.93): 20 C 78.38; H 9.48. Found: C 78.27; H 9.53.

Example 69 Preparation of cycloartenyl-3-ethoxy-4-hydroxycinnamate Cycloartenyl-4-acetoxy-3-ethoxycinnamate (30.3 g, 0.046 mole) prepared according to procedure of Example 68 was dissolved in tetrahydrofuran (300 ml), followed i by addition of 25 aqueous ammonia (60 ml). The mixture was stirred at 50 0 C for 2 hours. After the 4.,

P

iT~ -n i_ -i 146 1 reaction was complete, the mixture was evaporated to dryness under reduced pressure, Pnd the residual crystals were recrystallized from acetone-water (2:1, V/V) giving cycloartenyl-3-ethoxy- 4 -hydroxycinnamate (19.7 in a 69 yield. m.p. 134 135 0

C.

24 Specific rotation [a]D +40.80 (C 1.00, CHCl 3 Analysis, Calcd. for C 4 1

H

60 0 4 616.89): C 79.82; H 9.80. Found: C 79.89; H 9.73.

Example Preparation of cyclobranyl-4-acetoxy-3-ethoxycinnamate The title compound was prepared according to following the procedure of Example 68 vwere cyclobranol (33.1 g, 0.075 mole) was used as starting material; the yield was 43.1 g (85 m.p. 153 154 0

C.

24 Specific rotation [a]D +37.60 (C 1.00, CHC13) Analysis, Calcd. for C 44

H

6 4 0 5 672.95): C 78.53; H 9.59. Found: C 78.48; H 9.64.

Example 71 Preparation of cyclobranyl-3-ethoxy-4-hydroxycinnamate 20 The title compound was prepared according to following the procedure of Example 69 where cyclobranyl-4-acetoxy-3-ethoxycinnamate (30.9 g, 0.046 mole; obtained by the procedure of Example 70 was used; the yield was 22.8 g (78 m.p. 181 182 0

C.

24 Specific rotation [a]D +38.00 (C 1.00, CHCl 3 Analysis, Calcd. for C 42

H

62 0 4 630.92): C 79.95; H 9.91. Found: C 79.91; H 9.98.

t IC a a 4 CL 1 147 1 Example 72 Preparation of 24-methylenecycloartanyl-4-acetoxy- 3-ethoxycinnamate The title compound was prepared according to 1 following the procedure of Example 68 where 24methylenecycloartanol (33.1 g, 0.075 mole) was used as starting material; the yield was 42.7 g (84 m.p. 184 185 0

C.

24 Specific rotation [a]D +39.90 (C 1.00, CHCl 3 Analysis, Calcd. for C 4 4

H

6 4 0 5 672.95): C 78.53; H 9.59. Found: C 78.59; H 9.52.

r Example 73 P -eparation of 24-methylenecycloartanyl-3-ethoxy- 4-hydroxycinnamate The title compound was prepared according to following the procedure of Example 69 where 24methylenecycloartanyl-4-acetoxy-3-ethoxycinnamate (30.9 g, 0.046 mole) obtained according to the procedure of Example 72 was used; the yield was 23.4 g t" 20 (80 m.p. 146 147 0

C.

rtt 24 Specific rotation [a]D +40.10 (C 1.00, CHC3) Analysis, Calcd. for C 4 2

H

6 2 0 4 630.92): r« C 79.95; H 9.91. Found: C 79.89; H 9.95.

Example 74 Preparation of cycloartenyl-4-hydroxy-3-propoxycinnamate The title compound was prepared according to 14 1 following the procedure of Example 69 cycloartenyl-4acetoxy-3-n-propoxycinnamate (31.0 g, 0.046 mole) obtained according to the procedure of Example 68; the yield was 20.5 g (70 m.p. 144 145 0

C.

24 Specific rotation [a]D +40.20 (C 1.00, CHC1 3 Analysis, Calcd. for C 42

H

62 0 4 630.92): C 79.95; H 9.91. Found: C 79.99; H 9.85, Example Preparation of cyclobranyl-4-hydroxy-3-propoxycinnamate The title compound was prepared according to following the procedure of Example 69 being used cyclobranyl-4-acetoxy-3-propoxycinnamate (31.6 g, 0.046 mole) obtained according to the procedure of Example 68; the yield was 22.8 g (76 m.p. 187 188 0

C.

24 Specific rotation [a]D +37.40 (C 1.00, CHCl 3 Analysis, Calcd. for C 43

H

64 0 4 644.94): C 80.07; H 10.00. Found: C 79.92; H 10.13.

Example 76 20 Preparation of cycloartenyl-3,4-dimethoxycinnamate Acetone (200 ml) and potassium carbonate 1 (10.0 g) were added to cycloartenyl-4-hydroxy-3- *methoxycinnama'te (10.0 g, 0.017 mole) at 20 0 C and the mixture was stirred. Thereto was added dimethyl sulfate (2.8 ml, 5 equivalents) and continued to stir I at 50°C for 3 hours. Then the potassium carbonate was separated by filtration, and the filtrate was 149 1 evaporated under reduced pressure. The residue was dissolved in methylene chloride, and the methylene chloride layer washed with saturated aqueous solution of sodium bicarbonate. The aqueous layer was extracted with methylene chloride and the combined extracts were dried, evaporated to dryness under reduced pressure, and the residue was recrystallized from methylene chloride-methanol giving the title compound (9.6 g) in a 94 yield. m.p. 136 137 0

C.

23 Specific rotation [a]D +41.70 (C 1.03, CHC1 3 Analysis, Calcd. for C 4 1

H

6 0 0 4 616.89): C 79.82; H 9.80. Found: C 79.90; H 9.88.

Example 77 Preparation of cycloartenyl-3-methoxy-4-propionyloxycinnamate Pyridine (160 ml) was added to cycloartenyl- 4-hydroxy-3-methoxycinnamate (40.0 g, 0.066 mole) and the mixture was stirred at 0 C. Thereto was added propionic anhydride (80 ml) dropwise and the solution 20 was continued to stir at 20 0 C for 1 hour, then allowed to stand at 5 0 C for 48 hours. After the reaction was completed water (250 ml) was added to the solution to precipitate crystals of the title compound, which

S

were then filtered and washed with water and methanol.

The yield was 42.3 g (95 m.p. 156 157 0

C.

Specific rotation [a] 2 5 +40.20 (C 1.04, CHC1 3 Analysis, Calcd. for C 4 3 H6205 658.93): SC 78.38; H 9.48. Found: C 78.45; H 9.42.

Example 78 Preparation of cycloartenyl-3,4-dimethoxybenzoate Toluene (10 ml), thionyl chloride (48 ml, equivalents) and pyridine (1.0 ml) were added to 3,4-dimethoxybenzoic acid (11.8 g, 0.065 mole) at 0°C, and the mixture was stirred at 40 0 C for 10 minutes.

The reaction mixture was evaporated under reduced pressure, and to the residue were added cycloartenol (20.0 g, 0.043 mole) and pyridine (100 ml) at 0°C.

so> The mixture was heated to 40°C and stirred for 1 hour 0,O*o and furthermore continued to stir at 60°C for 1 hour.

2 The resulting mixture was evaporated under reduced o 0 9 o pressure, and the residual crystals were dissolved in 15 chloroform. The organic layer was washed with saturated aqueous solution of sodium bicarbonate, then aqueous layer was extracted with chloroform. The combined 0 16 4* extracts were dried, and evaporated under reduced pressure. The resulting residue was purified by silica i 20 gel column chromatography (solvent: toluene-methylene chloride, giving cycloartenyl-3,4dimethoxybenzoate (25.9 in a 88 yield- m.p.

147 147.5 0

C.

26 Specific rotation [a]D +63.80 (C 1.00, CHCl 3 Analysis, Calcd. for C 3 9

H

5 8 0 4 590.85): C 79.27; H 9.89. Found: C 79.34; H 9.82.

151.

1 Example 79 Preparation of cyclobranyl-3,4-dimethoxycinnamate To 3,4-dimethoxycinnamic acid (16.53 g, 0.0794 mole) suspended in toluene (200 ml) was added thionyl chloride (11.5 ml, 2 equivalents) and the mixture was stirred at 100 0 C for 2.5 hours. The resulting mixture was evaporated under reduced pressure, and the residue was suspended in a toluene (100 ml) and pyridine ml). To the suspension was added cyclobranol (17.5 g, 0.0397 mole) and the mixture was stirred at 100 0 C for 3 hours. The solution was evaporated under reduced S. pressure, and the residue was extracted with chloroform (300 ml). The extract was dried, concentrated in vacuo, and purified by silica gel column chromatography (solvent: toluene), giving cyclobranyl-3,4dimethoxycinnamate (22.3 in a 93 yield. m.p.

168 169 0

C.

25 Specific rotation [a]D +38.50 (C 1.00, CHC1 3 Analysis, Calcd. for C42H6204 630.92): 20 C 79.95; H 9.91. Found: C 80.02; H 9.84.

Example Preparation of cyclobranyl-3,4 -dimethoxybenzoate a t To veratric acid (15.5 g, 0.085 mole) dissolved in toluene (150 ml) was added thionyl chloride (12.4 ml, 2 equivalents) and the mixture was stirred at 100°C for 1.5 hours. The solution was evaporated under reduced pressure, and the residue was dissolved in 1 toluene (100 ml) and pyridine (50 ml). To the solution was added cyclobranol (25 g, 0.057 mole) and the mixture was stirred at 60 0 C for 2 hours. The reaction mixture was evaporated to dryness under reduced pressure-, and the residue was extracted with chloroform (300 ml). The extract was dried, concentrated in vacuo, and purified by silica gel column chromatography (solvent: toluene), giving cyclobranyl-3,4dimethoxybenzoate (25.6 in a 76 yield. m.p.

158 159 0

C.

26 .4*4 Specific rotation [a]26 +59.20 (C 0.89, CHC1 3 *4 Analysis, Calcd. for C 40

H

60 0 4 604.88): C 79.42; H 10.00. Found: C 79.34; H 10.12.

0 Examples 81 100 15 The esters of 24-methylenecycloartanol in Examples 81 100 were prepared according to following the corresponding procedures of the cyclobranol's mentioned above. In these procedures the same amounts (by mole) of 24-methylenecycloartanol, in place of cyclobranol, was used. Yields melting points (OC), and specific rotations (Cl.00, CHC1 3 of these compounds are summarized in Table 13.

1 Table 13 Examnple Original organic acid Yield M, P Specif ic No. of ester M% rotation Ma 24

D

81 3,4-Diacetoxycinnanic 62 14.3-145 +36.20 acid 82 3,4-Dihydroxycinnamic 73 236-237 +42.50 acid 83 p-Acetoxycinnatic acid 76 158-159 +42.00 84 p-Hydroxycinnanic acid 87 240-241 +43.50 85 4-Acetoxy-3-inethoxy- 76 162-163 +58.20 benzoic acid 024086 4-Hydroxy-3-methoxy- 80 155-156 +60.20 benzoic acid 87 3,4-Diacetoxybenzolic 62 153-154 +52.30 acid 88 3,4-Dihydroxybenzoic 85 205-206 +61.40 acid 89 p-Nitrobenzoic acid 89 229-230 +61.80 p-Aminobenzoic acid 72 174-175 +60.20 0 91 Nicotinic acid 86 173-174 +66.50 V4#, 92 o-Hydroxybenzoic acid 69 161-162 '1 20 93 Linoleic acid 69 oily +38.10 m~atter 94 p-Hydroxybenzoic acid 92 177-178 +64.40 r-Acetoxybenzoic acid 90 133-134 +60.10 96 m-Hydroxybenzoic acid 85 185-186 +64.20 97 o-Nitrobenzoic acid 89 190-191 +92.50 98 o-Arninobenzoic acid 67 196-197 +76.10 99 r-Nitrobenzoic acid 85 170-171 +60.80 100 m-Aminobenzoic acid 82 178-179 +62.00 154 1 Example 100-1 Preparation of cycloartenyl-4-hydroxy-3-methoxycinnamate Methanol (350 ml) was added to a solution or y-oryzanol (1.0 kg, cycloartenol ester content 44 in acetone (5.0 1) at an elevated temperature. Then the mixture was cooled and allowed to stand overnight at 20 0 C. The thus separated crystals were filtered, giving y-oryzanol (680 g) in which the content of cycloartenyl ester was 60 Similar recrystallization of this y-oryzanol by using acetone (3.4 1) and methanol (68 ml) gave y-oryzanol (450 g) of cycloartenyl ester content 75 Further repeated recrystallizations the first time from acetone S 15 (2.25 the second ethyl acetate (1.25 the third acetone (1.29 and the fourth acetone (960 ml), respectively, yielded 250 g (88 purity on cycloartenyl ester), 184 g (92 purity), 120 g (97 Spurity) and 91 g (98 purity) of y-oryzanol, and the final recrystallization from ethyl acetate (640 ml) gave cycloartenyl-4-hydroxy-3-methoxycinnamate (53 g), in a 5.3 yield, m.p. 153 153.5 0

C.

2 Specific rotation t 2 1 .5 +41.20 (C 0.99, CHC1 3 Analysis, Calcd. for C 40

H

58 0 4 602.86): C 79.69; H 9.70. Found: C 79.73; H 9.62.

IRV, KBr 3400, 2910, 1700, 1672, 1599, 1510, 1270, 1155.

I155 1 PMR (CDC1 3 0.38 (1H, 1/2 ABq, 4.2 Hz), 0.58 (1H, 1/2 ABq, 4.2 Hz), 0.70-2.40 (27H, m), 0.89 (6H, 0.96 (6H, 1.61 (3H, bs), 1.68 (3H, bs), 3.91 (3H, 4.50-4.90 (1H, m), 4.80-5.30 (1H, 5.87 (1H, 6.28 (1H, 1/2 ABq, 15 Hz), 6.76-7.10 (2H, 7.00 (1H, 7.55 (1H, 1/2 ABq, 15 Hz).

Example 100-2 Preparation of cyclobranyl-4-hydroxy-3-methoxycinnamate Cyclobranyl-4-actoxy-3-methoxycinnamate (44.9 g, t ,t ,t 0.068 mole) prepared according to the procedure of Example 65 was dissolved in dioxane (900 ml), and to the solution was added 25 aqueous ammonia (90ml) dropwise. After the mixture was stirred at 50 0 C for 2 hours, the solution was evaporated to dryness under reduced pressure. The residual crystals were washed with ethanol, giving cyclobranyl-4-hydroxy-3-methoxycinnamate (40.0 in a 95.2 yield. m.p. 191 192 0

C.

Specific rotation a)23.5 +38.50 (C 0.99, CHC1 3 Analysis, Calcd. for C41H6004 616.89): C 79.82; H 9.80. Found: C 79.77; H 9.88.

SIR, KBr (cm 3500, 2920, 2850, 1690, 1600, 1510, 1265, 1155.

PMR (CDC1 3 0.36 (1H, 1/2 ABq, 4.8 Hz), 0.60 (1H, 1/2 ABq, 4.8 Hz), 0.68-2.20 (27H, m), 0.89 (6H, 0.96 (6H, 1.62 (9H, s), 156 1 3.90 (3H, 4.50-4.90 (1H, 5.97 (1H, b), 6.27 (1H, 1/2 ABq, 15.6 Hz), 6.75-7.20 (3H, m), 7.9 (1H, 1/2 ABq, 15.6 Hz).

Example 100-3 Preparation of 24-methylenecycloartanyl-4-hydroxy-3methoxycinnamate Commercial available y-oryzanol (100 g, 24methylenecycloartanol content 45 was acetylated with acetic anhydride in pyridine. This acetylated y-oryzanol was recrystallized repeatedly from chloroform-ethyl acetate-ethanol V/V), giving acetyl ferulate (18 g) of 24-methylenecycloartanol content 95 This acetyl ferulate was completely saponified in the solution of 2N NaOHethanol, then the obtained alcohol was converted into C t t the benzoate. This benzoate was recrystallized repeatedly, and completely saponified'to give 24to methylenecycloartanol (5 A portion (2g, 0.0045 mole) thereof was converted into 24-methylenecycloartanol-4-acetoxy-3-methoxycinnamate according to following the procedure of Example 65. This ester 4 #4 was then deacetylated according to following the procedure of Example 100-2, giving 24-methylenecycloartanol-4-hydroxy-3-methoxycinnamate (2.58 in a 25 90 yield. m.p. 166 167 0

C.

24 S* *Specific rotation [a]D +40.60 (C 1.00, CHCl 3 Analysis, Calcd. for C 41 H 6004 1/3C2 5

O

17 It 632.25): C 79.15; H 9.88. Found: C 79.11; H 9.94.

IRV, KBr (cm- 3400, 2920, 2850, 1688, 1630, 1600, 1510, 1463, 1425, 1265, 1158 PMR (CDC1 3 0.39 (1H, .1/2 ABq, 4.2 Hz), 0.59 (1H, 1/2 ABgj, 4.2 Hz), 0.60-2.50 (28H, 0.82 (6H, s) 0.95 (6H, s) 1.02 (6H, a, 7.2 Hz), 3.91 (3H, s) 4.70 (2H, bs) 4.50-4.90 (1H, m), 5.95 (1H, s) 6.70-7.30 (3H, mn), 6.32 (1H, 1/2 ABq, 16 Hz), 7.56 (1H, 1/2 ABq, 16 Hz).

e.

S

till 4* 44 I I p St ItO

I

00 0* to It I I

I

I P Si

I

It 00

I

ISO I ~1 0000 0 0.1.4*o *J *J a

I

1-; 158 1 Example 101 Preparation of cycloartenyl ester of p-nitro cinnamic acid Thionyl chloride (112 ml, 4 equivalents) and dimethylformamide (1 ml) were added to p-nitrocinnamic acid (73.0 g, 0.378 mole) and the mixture was stirred at 60 0 C for 2 hours. Then the resulting mixture was concentrated under reduced pressure.

Dioxane (250 ml), pyridine (250 ml), and then cycloartenyl (125 g, 0.293 mole) were added to the residue. After 2 hours' stirring at 60 0 C, the solvents were removed by distillation under reduced pressure, and the residue was extracted with chloroform. The extracts were washed successively with water and t ,t saturated aqueous NaHCO 3 then dried, and concentrated under reduced pressure. The residual crystals were recrystallized from methylene chloride-methanol cycloartenyl ester of p-nitrocinnamic acid (158.0 in a 89.6 yield. m.p. 199 200 0

C.

26 Specific rotation [a]D 43.80(C1.00, CHCl).

S 20 Analysis, Calcd. for C39H55NO4 (M.W.601.83): C,77.83;

C

4 H,9.21; N,2.33. Found: C,77.89; H,9.16; N,2.28.

t t, IRv, KBr(cm 2930, 1708, 1640, 1600, 1520, 1345, 1205, 1175.

PMR(CDC1 3 0.38(1H, 1/2ABq, 4.2Hz), 0.61(lH, 25 1/2ABq, 4.2Hz), 0.52-2.36(27H, 0.91(6H, s), 25" 0.98(6H, 1.61(3H, 1.63(3H, s), 4.83-4.90(1H, 4.90-5.31(1H, 6.55(1H, 153 1 1/2ABq, 16.2Hz), 7.65(2H, ABq, 8.7Hz), 7.67 (1H, 1/2ABq, 16.2Hz), 8.24(2H, ABq, 8.7Hz).

Example 102 Preparation of cycloartenyl ester of p-amino-cinnamic acid Cycloartenyl ester of p-nitro cinnamic acid (160.0 g, 0.266 mole) prepared according to the procedure of Example 101 was suspended in a mixture of acetic acid (1.5 Z and dioxane (1.5 To the suspension were added 6N-HCz-dioxane (95 mu) and zinc powder (80 g) and the mixture was stirred at 40 0 C for 3 hours. After th reaction, zinc powder was removed by filtration. The filtrate was concentrated under o reduced pressure, and the residue was extracted with chloroform. The extracts were washed successively with Swater and saturated aqueous NaHCO 3 then dried, and 0 concentrated under reduced pressure. The residual crystals were recrystallized from chloroform-ethanol giving cycloartenyl ester of p-aminocinnamic acid (140.0 in a 92.0 yield, m.p. 185 187 0

C.

tr; t 26 Specific rotation [a] D 42.3(C 1.00, CHCl 3 43 Analysis, Calcd. for C 3

H

57

NO

2 (M.W.571.85): C,81.91; 3 57 239H70 S. H,10.05; N,2.45. Found: C,81.87; H,10.03; N,2.53.

IRv, KBr(cm-1): 3450, 3350, 2920, 1695, 1620, 1600, 5 1515, 1440, 1205, 1165.

0 PMR(CDC1 3 0.38(iH, 1/2ABq, 4.2Hz), 0.61(1H, 1/2ABq, 4.2Hz), 0.72-2.80(27H, 0.91(6H, 0.96(6H s), 160 1 1.60(3H, 1.68 (3H, 3.92(2H, bs), 4.48- 4.88(1H, 4.88-5.32(1H, 6.22(1H, 1/2ABq, 15.8Hz), 6.63(2H, ABq, 8.4Hz), 7.35(2H, ABq, 7.56(1H, 1/2ABq, 15.8Hz).

Example 103 Preparation of cyclobranyl ester of pnitrocinnamic acid Thionyl chloride (60 mt, 2 equivalents),dioxane (300 mz), and dimethylformamide (1 ma) were added to p-nitrocinnamic acid (75 g, 0.388 mole), and the mixture was stirred at 60°C for 2 hours. Then the rersulting mixture was concentrated under reduced pressure. To the residue were added dioxane (300 mz), pyridine (200 mt) and then cylobranol (130 g, 0.295 mole). After 2 hours' stirring at 60 0 C, the solvents were removed by distillation under reduced pressure, and the residue was extracted with chloroform. The o99o extracts were washed successively with water and saturated aqueous NaHCO 3 then dried, and concentrated 20 in vacuo. The residue was recrystallized from ct chloroform-ethanol giving cyclobranyl t. cr ester of p-nitrocinnamic acid (168.4 in a 92.7 yield. m.p. 231 232 0

C.

Specific rotation []26 40.6 0 (C 1.00, CHC1 3 j 25 Analysis Calcd. for C40H57NO4(M.W.615.86): C,78.01; .9 H,9.33; N,2.27. Found: C,78.10; H,9.24; N,2.36.

IRv, KBr(cm 2930, 1710, 1635, 1600, 1520, 1345, f 'i 1G1 1 1300, 1175.

PMR(CDC1 0.36(1H, 1/2ABq, 4.8Hz), 0.61(1H, 1/2ABq, 4.8Hz), 0.68-2.20(27H, 0.91(6H, s), 0.97(6H, 1.62(9H, 4.48-4.92(1H, m), 6.55(1H, 1/2ABq, 15.6Hz), 7.67(2H, ABq, 8.4Hz), 7.69(1H, 1/2ABq, 15.6Hz), 8.22(2H, ABq, 8.4Hz).

Example 104 Preparation of cyclobranyl ester of paminocinnamic acid Cyclobranyl ester of p-nitrocinnamic acid (165.0 g, 0.268 mole) prepared according to the procedure of Example 103 was suspended in a mixture of acetic acid (1.5 A) and tetrahydrofuran (2 To the 15 suspension were added 6N-HCl-dioxane (125 mk) and zinc powder (165 g) and the mixture was stirred at 20 0 C for "2 hours. Then the reaction mixture was filtered to Is. remove zinc powder. The filtrate was concentrated under reduced pressure, and the residue was extracted with chloroform. The extracts were washed successively Swith water and saturated aqueous NaHCO 3 then dried, and t31 Q C I concentrated in vacuo. The residual crystals were S *recrystallized from chloroform-ethanol v/v), giving cyclobranyl ester of p-aminocinnamic acid 25 (139.2 in a 75.9 yield. m.p. 206 207 0

C.

Specific rotation [a] 2 6 40.1(C 1.00, CHCl3 Analysis Calcd. for C 40

H

59

NO

2 (M.W.585.87): C,82.00; 1 H,10.15; N,2.39. Found: C,81.92; H,10.14; N,2.43.

-1 IR'v, KBr(cm 3450, 3350, 2930, 1690, 1620, 1595, 1515, 1440, 1300, 1270, 1170.

PMR(CDCl 3 0.35(1H, l/2ABq, 4.8Hz), 0.59(lH, 1/2ABq,, 4.8Hz), 0.68-2.36(27H, in), 0.90(6H, s), 0.96(6H, 1.62(9H, 3.76-4.08(2H, in), 4.44-4.87(1H, mn), 6.22(lH, l.2ABq, 15.6Hz), 6.62(2H, ABq, 8.4Hz), 7.34(2H, ABq, 8.4Hz), 7.57(lH, 1/2ABq, 15.6Hz).

Example 105 Preparation of 24-methylenecycloartanyl ester of p-nitrocinnamic acid o P 00*4 The title compound was prepared according to following the procedure of Example 103 where 24methylenecycloartanol (130 g, 0.295 mole) was used as a starting material in place of cyclobraiol. The yield was 164.8 g (90.7 223 224'C.

Specific rotation [at] 26 43.10(0 1.00, CHC1 D 3 Analysis Calcd. for C 40

H

57 NO'(M.W.615.86): C,78.01; H,9.33; N,2.27. Found: C,78.08; H,9.25; N,2.34.

0* Pp Example 106 Preparation of 24-methylenecycloartanyl ester of p-ainncinnamic acid The title compound was prepared according to following the procedure of Example 104 where 24methylenecycloartanyl ester of p-nitrocinnanic acid 1 (163.5 g, 0.265 mole) as a starting material. The yield was 118.3 g (76.2 m.p. 201 202°C.

26 Specific rotation [a]D 42.00(C 1.00, CHC13 Analysis, Calcd. for C 4 0

H

5 9

NO

2 (M.W.585.87): C,82.00; H,10.15; N2.39. Found: C,81.95; H,10.19; N2.42.

Example 107 Preparation of cycloartenyl ester of mnitrocinnamic acid The title compound was prepared according to following the procedure of Example 101 but m-nitrocinnamic acid (73.0 g, 0.378 mole) was used as a starting material in place of p-nitrocinnamic acid.

The yield was 156.8 g (88.9 m.p. 181 182 0

C.

26 Specific rotation 42.40(C 1.00, CHC1) D 3 Analysis, Calcd. for C 3 9

H

5 5

NO

4 (M.W.601..83): C,77.83 H,9.21; N,2.33. Found: C,77.76; H,9.26; N,2.39.

Example 108 Preparation of cycloartenyl ester of m- 20 aminocinnamic acid 0 The title compound was prepared according to following the procedure of Example 102 but cycloartenyl ester of m-nitrocinnamic acid (160.0 g, 0.266 mole) 0 obtained according to the procedure of Example 107 was O 9l 4, 25 used in place of cycloartenyl ester. The yield was 135.8 g (89.2 m.p. 189 190 0

C.

26 43.1(C 1.00, CHC1 Specific rotation D 43.1 (C 1.00, CHC1 3 1i 4 1 Analysis Calcd. for C 39

H

57

NO

2 (M.W.571.85): C,81.91; H,10.05; N,2.45. Found: C,81.87; H,10.08; N,2.53.

Example 109 Preparation of cyclobranyl ester of m-nitrocinnamic acid The title compound was prepared according to following the procedure of Example 103 but m-nitrocinnamic acid (75 g, 0.388 mole) was used as a starting material. The yield was 167.6 g (92.2 m.p.204 205 0

C.

Specific rotation [a]26 40.50(C 1.00, CHC1 3 Analysis Calcd. for C 40

H

57

NO

4 (M.W.615.86): C,78.01; i t H,9.33; N,2.27. Found: C,77.95; H,9.38; N,2.32.

Example 110 Preparation of Cyclobranyl ester of m-aminocinnamic acid The title compound was prepared according to following the procedure of Example 104 but cyclobranyl S. 20 ester of m-nitrocinnamic acid (163.2 g, 0.265 mole) obtained according to the procedure of Example 109, S, was used as a starting material. The yield was 117.4 g S (75.6 m.p.211 212 0

C.

26 Specific rotation []26 41.20(C 1.00, CHCl S D 3 a l 25 Analysis Calcd. for C40H59NO2 (M.W.585.87): C,82.00; H,10.15; N,2.39. Found: C,82.07; H,10.08; N,2.43.

i-c, 1G65 1 Example 111 Preparation of 24-methylene cycloartanyl ester of m-nitrocinnamic acid The title compound was prepared according to following the procedure of Example 103 but m-nitrocinnamic acid (75 g, 0.388 mole) and 24-methylenecycloartanol (130 g, 0.295 mole) was used in place of p-nitrocinnamic acid and cyclobranol, respectively.

The yield was 167.2 g (92.0 m.p.193 194 0

C.

Specific rotation [a26 42.3°(C 1.00, CHC1 Specific rotation [c]D 3 Analysis Calcd. for C40H57NO (M.W.615.86): C,78.01; H,9.33; N,2.27. Found: C,78.05; H,9.27; N,2.34 Example 112 Preparation of 24-methylenecycloartanyl t ester of m-aminocinnamic acid S 15 The title compound was prepared according to t 6 following the procedure of Example 104 but 24-methylenetit ttt cycloartanyl ester of m-nitrocinnamic acid (160.4 g, 0.260 mole) obtained according to the procedure of Example 111 was used as a starting material. The yield 'r 20 was 114.8 g (75.3 m.p. 197 198 0

C.

0% 26 t Specific rotation [a6D 42.7°(C 1.00, CHC1 3 e '1 Analysis Calcd. for C 40H NO (M.W.585.87): C,82.00; H,10.15; N,2.39. Found: C,81.93; H,10.22; N,2.44.

tt Example 113 Preparation of cycloartenyl ester of 3methoxy-4-propionyloxy-a-methyl cinnamic acid 1 To 3-Methoxy-4-propionyloxy-a-methylcinnamic acid (72.0 g, 0.272 mole) were added thionyl chloride (40.0 mz, 2 equivalents), toluene (400 and dimethylformamide (0.5 mf) and the mixture was stirred at 60 0 C for 1.5 hours. After concentration of the mixture under reduced pressure, thereto dioxane (100 mt) was added and the whole was stirred at 0 C. To the mixture was added cycloartenol (80.0 g, 0.187 mole) dissolved in pyridine (300 mz) and the whole was stirred at 60 0 C for 3 hours. The reaction mixture was evaporated under reduced pressure and the residue was dissolved in chloroform (800 mt). The solution was washed with saturated aqueous NaHCO 3 and the aqueous layer was extracted with chloroform (500 mt x The combined chloroform layer was dried and evaporated under reduced pressure, and the residue t was purified by silica gel column chromatography [solvent hexane-methylene chloride, giving cycloartanyl ester of 3-methoxy-4-propionyloxy-a- 20 methylcinnamic acid (110 in a 87.1 yield. m.p.

130 131 0

C.

19 4 C 1.00 CHC1 Specific rotation 1 D +41.40(C 1.00, CHC13 Analysis Calcd. for C H 0 (M.W.672.95): C,78.53; 44 64 S H,9.59. Found: C,78.59; H,9.52.

25 IR), KBr(cm 2920, 2850, 1765, 1710, 1630, 1600, 1510, 1240, 1140, 1110.

PMR(CDC1 3 0.39(1H, 1/2ABq, 4.2Hz), 0.60(1H, '*3 L3i i -T°rrr- 1G7 1 1/2ABq, 4.2Hz), 0.60-2.20(27H, 0.90(6H, s), 0.98(6H, 1.27(3H, t, 7.2Hz), 1.58(3H, bs), 1.68(3H, bs), 2.12(3H, d, 1.2Hz), 2.62(2H, q, 7.2Hz), 3.80(3H, 4.50-5.30(2H, m), 6.80-7.70(4H, m).

Example 114 Preparation of cycloartenyl ester of 4-hydroxy-3-methoxy-a-methylcinnamic acid (another name: cycloartenyl ester of a-methylferulic acid) To cycloartenyl ester of 3-methoxy-4propionyloxy-a-methyl-cinnamic acid (84.0 g, 0.125 mole) prepared according to the procedure of Example 113 dissolved in dioxane (1000 mt) was added 25 aqueous ammonia (200 m) and the solution was stirred at 50 0

C

for 2 hours. Then, the reaction mixture was concentrated SS under reduced pressure to remove the solvent, and the residue was dissolved in chloroform. The chloroform solution was washed with saturated brine (500 mi), and 20 the aqueous layer were extracted with chloroform (300 mZ x The combined chloroform layer was dried, concentrated under reduced pressure, and the residue was recrystallized from methylene chloride-methanol gt giving cycloartenyl ester of 4-hydroxy- 25 3-methoxy-a-methylcinnamic acid (73.0 in a yield 94.8 m.p.143 1440C.

Specific rotation []D 1 9 44.10(C 1.00, CHC 3 M n 1%8 1 Analysis Calcd. for C41 H 6004(M.W.616.93): C,79.82; H,9.80.

Found: C,79.88; H,9.81.

IRv, KBr(cm-l): 3400, 2900, 2850, 1695, 1690, 1625, 1600, 1510, 1250, 1110.

PMR(CDC1 3 0.38(1H, 1/2ABq, 4.2Hz), 0.59(1H, l/2ABq, 4.2Hz), 0.60-2.30(27H, 0.88(6H, 0.97 (6H, 1.60(3H, bs), 1.66(3H, bs), 2.12(3H, d, 1.2Hz), 3.88(3H, 4.50-5.30(2H, m), 5.80(1H, bs), 6.70-7.70(4H, m).

Example 115 Preparation of cyclobranyl ester of 3-niethoxy-4-propionyloxy-a-methylcinnamic acid To 3-Methoxy-4-propionyloxy-a-methylcinnamic acid (15.59 g, 0.059 mole) suspended in toluene (50 mk) was added thionyl chloride (20 mZ, 4.6 equivalents) and dimethyl formamide (5 drops) and the mixture was stirred at 60 0 C for 2 hours. Then the solvent was removed by distillation under reduced pressure. The residue was suspended in toluene 20 (150 mZ) and anhydrous pyridine (30 mR), and to the suspension was added cyclobranol (20 g, 0.045 mole).

S The mixture was stirred at 60 0 C for 2 hours, then concentrated under reduced pressure. The residue was extracted with chloroform (300 mR), and the 25 chloroform solution was washed, dried, and evaporated to dryness under reduced pressure. The crystalline residue was washed with ethanol (50 mZ), and recrystal- L 1 lized from acetone-water (19:1, v/v) 1 giving cyclobranyl ester of 3-methoxy-4-prop2ionyloxy-amethylcinnamic acid (24.69 in a 79.2 yield.

m.p.146 147 0

C.

Specific rotation 19+ 39.20 (C 1.00, CHCl) D3 Analysis Calcd. for C 45

H

66 0 5 (M.W.686.98): C,78.67; H,9.68. Found: C,78.75; H9.62.

IRV, KBr(cm )1 3400, 2590, 2850, 1760, 1710, 1630, 1600, 1240, 1150, 1120.

PMR(CDC 3 )63: 0.37 (lH, l/2ABq, 4.8Hz), 0.62 (lH, .1/2ABg, 4.8Hz), 0.70-2.22(27H, in), 0.92(6H, s), 0.99(6H, 1.29(3H, t, 7.2Hz), 1.64(9H, s), 2.14(3H, d, 1.2Hz), 2.63(2H, q, 7.2Hz), 3.84 (3H, s) 4.48-4.88 (lE, mn), 6.80-7.08 (3H, m), 7. 59 (1H, q, 1. 2Hz).

Example 116 Preparation of cyclobranyl ester of 4hydroxy-3-methoxy-a-methylcinnanic acid Cyclobranyl ester of 3-methoxy-4-propionyloxy- 2a-methyl cinnamic acid (24.69 g, 0.036 mole) obtained according to the procedure of Example 115 was dissolved in dioxane (400 mz) and to the solution was added aqueous ammonia dropwise. The mixture was stirred at 50 0 C for 2 hours, then the solvent was removed by 2distillation under reduced pressure. The crystalline residue was washed with ethanol, and recrystallized from acetone-water (19:1, vlv), giving cyclobranyl 17 0 1 ester of 4-hydroxy-3-methoxy-a-methylcinnamic acid (21.72 g)in a 95.8 yield. in.p. 185 186'C.

Specific rotation [aL] 20 43.7"(C 1.00, CHCL 3 Analysis Calcd. for C 42

H

2 0 4 (M.W.630.92): C,79.95; H,9.91. Found: ,C,79.90; H,9.98.

KBr cm 1 3380, 2920, 2850, 1693, 1600, 1510, 1285, 1250, 1120.

PMR(CDC1 0.36(lH, 1/2ABq, 4.8Hz), 0.6l(lH, 1/2ABq, 4.8Hz), 0.76-2.30(27H, m) 0.91(6H, s), 0.99(6H, 1.63(9H, 2.14(3H, d, 1.2Hz), 3.90(3H, s) 4.48-4.84 (lH, m) 5.84(1H, bs), 6.80-6.98 (3H, m) 7.55(111, q, 1.2Hz).

*09 Example 117 Preparation of 24-methylenecycloartanyl 00 6 cinnamic acid 0 00 To 3-methoxy- 4-propionyloxy-a-rnethylcinnamic acid (0.8 g, 0.003 mole) susp~ended in toluene (2 mt) was added thionyl chloride (0.5 mR., 2.2 equivalent) and :00 20 dimethylformamide (2 drops), and the mixture was 0 stirred at 60 0 C for 2 hours. Then the solvent was removed by distillation under reduced pressure. The residue was suspended in toluene (2 ms.) and anhydrous 0 pyridine (1 and to the suspension was added 24-methylene cycloartanyl (1g, 0.0023 mole). The mixture was stirred at 60 0 C for 2 hours, then the solvents were removed by distillation under reduced 171 1 Pressure. The residue was extracted with chloroform mYZ) and the chloroform extract was washed with sodium bicarbonate,dried, and evaporated under reduced pressure. The crystalline residue was washed with ethanol (5 mZ) and recrystallized from acetonemethanol v/v) giving 24-methylenecycloartanyl ester of 3-methoxy-4-propionyloxy-a-methylcinnamic acid 19 Specific rotation D± 41.20 (C 1.00, CHCl 3 Analysis Calcd. for C H 0 (M.W.686.98): C,78.67; H,9.68. Found: C,78.75; H,9.62.

IRV, KBr(cm ):3400, 2920, 2850, 1760, 1700, 1240, 1115.

PMR(CDC1 3 0.36(lH, l/2ABg, 4.2Hz), 0.61(lH, 1/2ABg, 4 .2Hz) 0 .7 0-2 .22 (34H, m) 0. 88 s) it'4 0.96(6H, 1.26(3H, t, 7.2Hz), 2.11(3H, d, 1.2Hz), 2.60(2H, q, 7.2Hz), 3.80(3H, s), 4.44-4.86(lH, in), 4.86-5.26(2H, in), 6.76-7.08 (3H, mn), 7.55(lH, q, 1.2Hz).

Example 118 Preparation of 24-methylenecycloartanyl ester of 4-hydroxy-3-methoxy-a-rnethylcihnamic acid 24-Methylenecycloartanyl ester of 3-inethoxy- 4-propionyloxy-a-methylcinnamic acid (1.35 g, 0.002 mole) obtained according to the procedure of Example 117 was dissolved in aioxane (20 in94, and to the solution was 172 1 added 25 aqueous ammonia (2 mf) dropwise. The mixture was stirred at 500C for 2 hours, then the solvents were removed by distillation under reduced pressure.

The crystalline residue was washed with ethanol, and recrystallized from ethanol, giving 24-methylenecycloartanyl-ester of 4-hydroxy-3-methoxy-a-methylcinnamic acid (1.02 in a 82.2 yield. m.p. 144 145 0

C.

20 Specific rotation [a]D 0 44.8(1.00, CHC1) Analysis Calcd. for C43H6204(M.W.630.92): C,79.95; H,9.91. Found: C,79.99; H,9.84.

IRv, KBr(cm- 1 3400, 2900, 2850, 1690, 1600, 1510, 1250, 1110.

PMR(CDC1 3 0.37(1H, 1/2ABq, 4.2Hz), 0.61(1H, 1/2ABq, 4.2Hz), 0.70-2.21(34H, 0.89(6H, 0.98 (6H, 2.14(3H, d, 1.2Hz), 3.88(3H, s), 4.50-4.88(1 H, 4.88-5.28(2H, 5.80(1H, bs), 6.82-7.10(3H, 7.59(1H, q, 1.2Hz) Example 119 Preparation of cycloartenyl ester of 4butyryloxy-3-methoxy-a-ethylcinnamic acid Thionyl chloride (15.0 m, 3.3 equivalents) was S. added dropwise to a solution of 4-butyryloxy-3methoxy-a-ethylcinnamic acid (18.0 g, 0.062 mole) in benzene (40 m) at 0°C, and the mixture was heated to S 25 r i t 60 0 C and allowed to stirr for 2 hours. Then the excess thionyl chloride and the solvent wer.e removed.

1 by distillation under reduced pressure. Pyridine m) and dioxane (40 mt) were added to the residue.

While cooling the mixture at 0°C, a solution of cycloartenol (17.5 g, 0.041 mole) in pyridine (30 mk) was added dropwise. This reaction mixture was allowed to stir overnight at 20 0 C. Then the solvents were removed by distillation under reduced pressure. The residue was extracted with chloroform (200 mk) and the extract was concentrated in vacuo. The residue was recrystallized from acetone-methanol v/v), giving cycloartenyl ester of 4-butyryloxy-3-methoxy-aethylcinnamic acid (22.4 g) in a 77.9 yield.

m.p. 118.5 119.5 0

C.

20 Specific rotation [a]D 35.7 0 (C 1.00, CHC1 3 Analysis Calcd. for C4 H O (M.W.701.00): C,78.81; 46 68 H 9.78. Found: C,78.72; H 9.86.

IRv, KBr(cm- 1 3400, 2920, 2800, 1700, 1600, 1510, 1230, 1120.

PMR(CDC1 0.36(1H, 1/2ABq, 4.2Hz), 0.52-2.26(29H, m), 20 0.61(1H, 1/2ABq, 4.2Hz), 0.90(6H, 0.96 (6H, 1.04(3H, t, 7.2Hz), 1.18(3H, t, 7.2Hz), 1.60(3H, 1.66(3H, 2.26-2.82(4H, m), 3.79(3H, 4.50.-4.88(1H, 4.88-5.28(1H, m), 6.70-7.12(3H, 7.48-7.68(1H, m).

t Example 120 Preparation of cycloartenyl ester of 4-hydroxy-3-methoxy-a-ethylcinnamic acid c 1174 1 Cycloartenyl ester of 4-butyryloxy-3-methoxy-aethylcinnamic acid (22.0 g, 0.0314 mole) obtained according to the procedure of Example 119 was dissolved in dioxane (200 mZ)4, and to the solution was added 25 aqueous ammonia,(20 m.Z) dropwise. The mixture was heated at 501C for 5 hours. Then the solvent was removed by distillation under reduced pressure. The residue was extracted with chloroform (200 mZ) and the extract was concentrated undor in vacuo. The residue was recrystallized from acetone-methanol giving cycloartenyl ester of 4-hydroxy-3methoxy-oa-ethylcinnamic acid (17.2 in a 86.8 yield.

m.p. 136 137'C.

Specific rotation [ai] 20 41.5- (C 1.00, CHCl) D3 Analysis Calcd. for C 42H 620 4(M.W.630.92): C,79.95; H,9.96. Found: C,79.90; H,9.83.

IRv, KBr(cm- 3400, 2830, 1700, 1595, 1510, 1240, 1120.

PMR(CDC 3 0.35(lH, l/2ABq, 4.2Hz), 0.50-2.18(27H, m), 0.60(lH, l/2ABq, 4.2Hz), 0.89(6H, 0.95(6H, s), 1.19(3H, t, 7.2Hz), 1.57(3H, 1.65(3H, s), 2.56(2H, bq, 7.2Hz), 3.87(3H, 4.47-4.85(lH, in), 4.85-5.24(lH, in), 5.76(lH, bs), 6.96-7.09(3H, in), 7.24-7.64(lH, in).

Example 121 Preparation of cyclobranyl ester of 4butyryloxy-3-methoxy-ca-ethylcilnamic acid e e t 3! t'~t 0 17 1 Thionyl chloride (15.0mt 3.3 equivalents) was added dropwise to a solution of 4-butyryloxy-3methoxy-a-ethylcinnamic acid (18.0 g, 0.062 mole) in benzene (40 mi) at 0°C, and the mixture was heated at 60 0 C for 2 hours. Then the excess thionyl chloride and the solvent were removed by distillation under reduced pressure. Pyridine (40 ml) and dioxane (40 mf) were added to the residue. While cooling the mixture at 0°C, cyclobranol (18.1 g, 0.041 mole) was added, then the mixture was warmed to 20 0 C and allowed to stir overnight. The solvents were removed by distillation under reduced pressure and the residue was extracted with chloroform (200 mR The extract was concentrated in vacuo, and the residue was recrystallized from acetone-ethanol giving cyclobranyl ester of 4-butyryloxy-3-methoxy-a-ethyl-cinnamic acid (22.3 in a 76.0 yield. m.p. 138 1390C.

Specific rotation [a] 2 0 33.7 0 (C 1.00, CHCl 3 2ialysis Calcd. for C 4 7

H

7 0 05 (M.W.715.03): C,78.94; 20 H,9.87. Found: C,78.89; H,9.88.

,IRv, KBr(cm 3400, 2920. 2850, 1760, 1710, 1625, I 1510, 1230, 1120.

PMR(CDC3 0.36(1H, l/2ABq, 4.8Hz), 0.52-2.22(29H, m), #tw 0.61(1H, 1/2ABq, 4.8Hz), t 25 0.92(6H, 0.97(6, 1.03(3H, t, 7.2Hz), 1.18(3H, t, 7.2Hz), 1.60(9H, 2.22-2.82 (4H, 3.81(3H, 4.48-4.90(1H, m), 1 6.70-7.18(3H, 7.40-7.64(1H, m).

Example 122 Preparation of cyclobranyl ester of 4-hydroxy-3-methoxy-a-ethylcinnamic acid Cyclobranyl ester of 4-butyryloxy-3-methoxya-ethylcinnamic acid (21.3 g, 0.0298 mole) obtained according to the procedure of Example 121 was dissolved in dioxane (200 mt), and to the solution was added aqueous ammonia (20 mz) by dropwise. The mixture was heated to 50 0 C and stirred for 5 hours. Then the solvent was removed by distillation under reduced pressure and the residue was extracted with chloroform (200 mk). The extract was concentrated in vacuo, and the residue was recrystallized from ethanol to give cyclobranyl ester of 4-hydroxy-3-methoxy-a-ethylcinnamic acid (17.1 in a 88.9 yield. m.p. 156 157 0

C.

Specific rotation [a]20 37.7(C 1.00, CHCl 3 Analysis Calcd. for C43H 640 (M.W.644.94): C,80.07; H 10.00. Found: C,80.13; H 10.12.

-1 IRv, KBr(cm 3400, 2930, 1696, 1235, 1130.

PMR(CDCl 3 0.38(1H, 1/2ABq, 4.8Hz), 0.52-2.22(27H, m), t 0.62(1H, 1/2ABq, 4.8Hz), 0.91(6H, 0.97(6H, s), 1.21(3H, t, 7.2Hz), 1.62(9H, 2.57(2H, bq, js, 7.2Hz), 3.98(3H, 4.48-4.86(1H, m), S 25 5.78(1H, bs), 6.70-7.00(3H, 7.53(1H, m).

S'c Example 123 Preparation of cycloartenyl ester of 177 1 4-propionyloxy-c-methylcinnamic acid Thionyl chloride (18.1 mZ, 3.3 equivalents) and dimethylformamide (0.5 mZ) were added dropwise to a solution of 4-propionyloxy-a-methylcinnamic acid (17.6 g, 0.075 mole) in benzene (40 ma) at 0°C. The mixture was heated to 60 0 C and allowed to stir for 2 hours. Then the excess thionyl chloride and the solvent were removed by distillation under reduced pressure.

Dioxane (40 mf) and pyridine (10 ml) were added to the residue. While cooling themixture at 0°C, a solution of cycloartenol (21.3 g, 0.050 mole) in pyridine (40 mf) was added dropwise. This reaction mixture was warmed o to 20 0 C and allowed to stir overnight. Then the solvents tt, were removed by vacuum distillation, and the residue o S 3er 15 was extracted with chloroform (200 mf). The extract S was concentrated under reduced pressure, and the residue was recrystallized from acetone-ethanol (1:2, giving cycloartenyl ester of 4-propionyloxy-amethylcinnamic acid (27.0 g) in a 83.9 yield. m.p.

S 20 87 88 0

C.

t re Sc 19 Specific rotation [aD 45.9 (C 1.00, CHC1 3 t t Analysis Calcd. for C 4 3

H

6 2 0 4 (M.W.642.93): C,80.33; H 9.72. Found: C,80.31; H 9.79.

-1 1 IRv, KBr(cm 3400, 2920, 2850, 1760, 1700, 1260, 1215, 1115.

PMR(CDC1 3 0.36(1H, 1/2ABq, 4.2Hz), 0.52-2.20(27H, m), 0.61(lH, 1/2ABq, 4.2Hz), 0.89(6H, 0.97(6H, s), 1 1.25(3H, t, 7.2Hz), 1.57(3H, 1.65(3H, s), 2.10(3H, d, 1.2Hz), 2.58(2H, q, 7.2Hz), 4.28-4.84 (1H, in), 4.92-5.24(lH, in), 6.92-7.09(2H, in), 7.11-7.50(2H, in), 7.50-7.70(lH, in).

Example 124 Preparation of cycloartenyl ester of 4-hydroxy-a-methylcinnanic acid Cycloartenyl ester of 4-propionyloxy-a-methylcinnamic acid (27.0 g, 0.042 mole) obtained by the procedure of Example 123 was dissolved in dioxane (200 mPZJ, and to the solution was added 25 aqueous ammonia dropwise. The mixture was heated to 509C and allowed to stir for 2 hours.

Then the solvent was removed by distillation under reduced pressure, and the residue was extracted with chloroform (200 mZ). The extract was concentrated in vacuo, and the residue was recrystallized from ethanol, giving cycloartenyl ester of 4-hydroxy-a-methylcinnamic acid (20.5 in a 83.1 yield. m.p. 190 -191'C.

Specific rotation []19 45.80 (C 1.00, CHCl) I ~D3 4 4 20 Analysis Calcd. for C 40

H

58 0 3 (M.W.586.86): C,81.86; H 9.96. Found: C,81.77, H 9.99.

IRv, kBr(cm ):3400, 2992, 2985, 1700, 1675, 1600, 1510, 1260, 1200, 1170.

0*06PMR(CDCl 3 0.36(l1*, l/2ABq, 4.2Hz), 0.52-2.24(27H, in), 25 0.61(lH, 1/2ABq, 4.2Hz), 0.90(6H, 0.98(6H, s), 1.6103H, 1.64(3H, 2.13 (3H, d, 1.2Hz), 4.50-4.88(1H, mn), 4.88-5.24(lH, in), 5.88-6.60 (1H, mn), 6.68-7.-12(2H, mn), 7.12-7.50(lH, in), 173 7.50-7.68(1H, m).

QO*9

D

O Q *I O 0B 0**o Example 125 Preparation of cyclobranyl ester of 4-propionyloxy-a-methylcinnamic acid Thionyl chloride (18.1 mt, 3.3 equivalents) and dimethylformamide (0.5 mz) were added dropwise to a solution of 4-propionyloxy-a-methylcinnamic acid (17.6 g, 0.075 mole) in benzene (40 mZ) at 0°C. The mixture was heated to 60 0 C and continued to stir for 2 hours. Then the excess thionyl chloride and the solvent were removed by distillation under reduced pressure. Dioxane (20 mZ) and pyridine (40 were added to the residue. While cooling the mixture at 0°C, cyclobranol (22.0 g, 0.050 mole) was added,then the mixture was warmed to 20 0 C and continued to stir overnight. Then, the solvents were removed by distillation in.vacuo, and the residue was extracted with chloroform (200 mZ). The extract was concentrated under reduced pressure, and the residue was recrystallized 20 from acetone-methanol giving cyclobranyl ester of 4-propionyloxy-a-methylcinnamic acid (26.3 g), in a 80.0 yield. m.p. 107 108 0

C.

19 34 (C 100, Specific rotation [a]D 34.7(C 1.00, CHCl 3 Analysis Calcd. for C 44

H

64 0 4 (M.W.656.95): C,80.44; H 9.82.

Found: C,80.39; H 9.77.

IRv, KBr(cm-1): 3400, 2920, 2850, 1860, 1710, 1630, 1260, 1200, 1165, 1120

I

*r 0 0* g* *0 4 r <.7 180 1 PMR(CDCl 3 0.36(1H, 1/2ABq, 4.8Hz), 0.61(1H, 1.2ABq, 4.8Hz), 0.74-2.32(27H, 0.89(6H, 0.96 (6H, 1.26(3H, t, 7.2Hz), 1.61(9H, 2.10 (3H, d, 1.2Hz), 2.58(2H, q, 7.2Hz), 4.46-4.86 (1H, 6.90-7.52(4H, 7.52-7.70(1H, m).

Example 126 Preparation of cyclobranyl ester of 4hydroxy-a-methylcinnamic acid Cyclobranyl ester of 4-propionyloxy-a-methylcinnamic acid (26.3 g, 0.040 mole) obtained according to the procedure of Example 125 was dissolved in dioxane (200 mf), and to the solution was added 25 aqueous ammonia dropwise. The mixture was heated to 50 0 C and continued to stir for 2 hours. Then the -tt i S' 15 solvent was removed by distillation under reduced r t pressure, and the residue was extracted with chloroform (200 mf). The extract was concentrated in vacuo, and the residue was recrystallized from acetone-methanol t o, giving cyclobranyl ester of 4-hydroxy-a- 20 methylcinnamic acid (20.7 in a 83.1 yield.

m.p. 203 204°C.

19 Specific rotation [aD 46.00(Cl.00, CHCl g Analysis Calcd. for C 41

H

60 0 3 (M.W.600.89) C,81.95; \r H,10,07. Found: C 81.99, H 10.07.

o ,cm- IRV, KBr(cm): 3400, 2920, 2850, 1780, 1605, 1510, 1265, 1200, 1170, 1125.

PMR(CDC1 3 0.36(LH, 1/2ABq, 4.8Hz), 0.61(H, 1/2ABq, (2H, 7.12-7.44(2H, 7.58(H, q, 1.2Hz).

Example 127 Preparation of 24-methylenecycloartanyl ester of 4-propionyloxy-a-methylcinnamic acid The title compound was prepared according to following the procedure of Example 125 but 24-methylenecycloartanol was used in place of cyclobranol. The yield was 25.8 g (78.5 m.p. 94 19 Specific rotation [a]D 44.20(C 1.00, CHC13) D 3 Analysis Calcd. for C44 H 640 (M.W.656.95): C,80.44; 1 .8H,9.82. Found: C,80.48; H 9.78.

4 0 Example 128 Preparation of 24-metnylenecycloartanyl ester of 4--hydroxy-a-methylcinnamic acid 24-Methylenecycloartanyl ester of 4-propionyloxya-methylcinnamic acid (24.0 g, 0.036 mole) obtained according to the procedure of Example 127 was dissolved in dioxane (200 mk) and to the solution was added aqueous ammonia dropwise. The mixturre was heated to 50 0 C and continued to stir for 2 hours. Then the solvent was removed by distillation under reduced 1 25 pressure, and the residue was extracted ith chloroform (200 ms). The extract was concentrated in vacuo and the residue was recrystallized from acetone-methanol the residue was recrystallized from acetone-methanol i i r 5 3 18:2 1 giving 24-methylenecycloartanyl ester of 4hydroxy-a-methylcinnamic acid (19.4 g) in a 89.6 yield.

m.p. 195 1960C.

Specific rotation [al 9 43.80 (C 1.00, CHC13) Analysis Calcd. for C 41

H

60 0 3 (M.W.600.89): C,81.95; H 10.07. Found: C,81.90; H 10.14.

Example 129 Preparation of cycloartenyl ester of 4butyryloxy-a-ethylcinnamic acid Thionyl chloride (4.8 mt, 5 equivalents) was added dropwise to a solution of 4-butyryloxy-aethylcinnamic acid (3.50 g, 0.0133 mole) in benzene (7 mt) at 0 C. The mixture was heated to 60 0 C and continued to stir for 2 hours. Then the excess S 15 thionyl chloride and the solvent were removed by I 45 4 distillation under reduced pressure. After addition of pyridine (10 mt) to the residue, a solution of cycloartenol (2.85 g, 0.0067 mole) in pyridine (10 m) was added to the mixture dropwise at 0°C. This mixture 20 was warmed to 20 0 C and continued to stir overnight.

t Then the solvent was removed by distillation under reduced pressure and the residue was extracted with chloroform (40 mt). The extract was concentrated in vacuo, and the residue was recrystallized from acetone-ethanol giving cycloartenyl ester of 4-butyryloxy-a-ethylcinnamic acid (3.63 g) in a 80.7 in a yield. m.p. 88 89 0

C.

i 120 1Specific rotation D 2 41.20 (C 1.00, CHC1 3 Analysis Calad. for C 45H 660 4(M.W.670.98): C,80.55; H 9.92. Found: C,80.64; H,9.84.

IRv, KBr(cin 3400, 2940, 2860, 1760, 1710, 1240, 1200, 1170, 1125.

PMR(CDCl 3 0.36(lH, 1/2ABq, 4.2Hz), 0.52-2.22(29H, in), 0.61(1H, 1/2ABq, 4.2Hz), 0.90(6H, 0.96 (6H, 1.03(3H, t, 7.2Hz), 1.18(3H, t, 7.2Hz), 1.58 (3H, s) 1.66 (3H, s) 2.22-2.80 (4H, m), 4.42-4.88(1H, in), 4.88-5.24(lH, mn), 6.89-7.18 (2H, mn), 7.18-7.46(2H, mn), 7.46-7.64(lH, in).

Example 130 Preparation of cycloartenyl ester of 4 4-hydroxy-ct-ethylcinnanic acid 4:15 Cycloartenyl ester of 4-butyryloxy-a-ethylacid (2.00 g, 0.003 mole) obtained according to the procedure of Examnple 129 was dissolved in dioxane (20 int4, and to the solution was added 25 aqueous ammonia (2 m2k) dropwise. Then the mixture was warmed to 50 0 C and continued to stir for 5 hours. The solvent was removed by distillation under reduced 4 pressure, and the residue was extracted with chloroform ins). The extract was concentrated in vacuo, and the residue was recrystallized from acetone, giving cycloartenyl ester o- 4-hydroxy-a-ethylcinnamic acid (1.68 in a 93.2 yield. in.p. 162.5 163 0

C.

Specific rotation (a 0+ 46.10 (C 1.00, CHCl 3 1 Analysis Calcd. for C H 0 (M.W.600.89): C,81.95; 41 60 3 H,10.07. Found: C,81.88; H 10.12.

IRv, KBr(cm 3300, 2920, 2800, 1760, 1710, 1625, 1500, 1280, 1240, 1200, 1165, 1120.

PMR(CDC 3 0.36(lH, l/2ABq, 4.2Hz), 0.52-2.22(27H, in), 0.61(lH, l/2ABq, 4.2Hz), 0.90(6H, 0.96(6H, s), 1.19(3H, t, 7.2Hz), 1.60(3H, 1.67(3H, s), 2.57(2H, bg, 7.2Hz), 4.47-4.88(1H, in), 4.92- 5.32(lH, in), 6.43-6.67(lH, in), 6.68-7.04(2H, in), 7.12-7.48 (2H, m) 7.52-7.69 (lH, m).

Example 131 Preparation of cyclobranyl ester of 4butyryloxy-a-ethylcinnaiic aci.(, Thionyl chloride (7.3 mz, 5 equivalents) was added dropwise to a solution-r of i!-butyryloxv-a-ethylcinnainic acid (5.25 g, 0.02 mole) in benzene (10 mZ) at 0 0 C. The mixture was heated to 60 0 C and allowed to stir for 2 hours, Then the excess thionyl c'hloride and *the solvent were rnrr. by distillation under reduced pressure. After ad-dition of pyridine (40 mg) to the residue, cyclobran-il (4.41 g, 0.001 mole) was added while cooling the mixture at 0 0 C. The mixture was .~4j warmed to 201C and continued to stir overnight. Then the solvent was removed by distillation =nder reduced pressure, and the residue was extracted with chloroform iL) The extract was concentrated in vacuo,~ and the residue was recrystallized from acetone-ethanol 18 1 giving cyclobranyl ester of 4-butyryloxya-ethylcinnamic acid (4.80 in a 70.1 yield.

m.p. 117.5 118 0

C.

20 Specific rotation [a"D 38.6°(C 1.00, CHC1 3 Analysis Calcd. for C46 H804(M.W.685.00): C,80.65; H,10.01. Found: C,80.59; H 10.06.

IRV, KBr(cm 3400, 2900, 2850, 1770, 1710, 1625, 1510, 1230, 1120.

PMR(CDC13) 6 0.36(1H, 1/2ABq, 4.8Hz), 0.52-2.24(29H, m), 0.61(1H, 1/2ABq, 0.90(6H, 0.96(6H, s), 1.03(3H, t, 7.2Hz), 1.18(3H, t, 7.2Hz), 1.59 (9H, 2.24-2.82(4H, 4.48-4.84(lH, m), 6.90-7.18(2H, 7.18-7.48(2H, 7.48-7.68(1H, m) Example 132 Preparation of cyclobranyl ester of 4hydroxy-a-ethylcinnamic acid Cyclobranyl ester of 4-butyryloxy-a-ethylcinnamic acid (4.11 g, 0.0060 mole) obtained according to the procedure of Example 131 was dissolved in dioxane It (30 mz), and to the solution was added 25 aqueous ammonia (3 mA) dropwise. The mixture was heated to 50 0 C and allowed to stir for 5 hours. Then, the solvent i i- was removed by distillation under reduced pressure, and the residue was extracted with chloroform (40 mk). The i 25 extract was concentrated in vacuo, and the residue was recrystallized from acetone, giving cyclobranyl ester of 4-hydroxy-a-ethylcinnamic acid (3.39 in a 91.9 1yield. in.p. 202 -203'C.

AnalsisCalcd. frC4 203(..1.2:C8.3 H-,10.16. Found: C,81.97; H,10.18.

KBr(cin) 3350, 2920, 2860, 1680, 1600, 1510, 1275, 1245, 1200, 1170, 1130.

PMR(CDC1 0.36(lH, l/2ABq, 4.8Hz), 0.52-2.24 (27H, mn), 0.61(lH, l/2ABq, 4.8Hz), 0.90(6H, s), 0.97(6H, 1.19(3H, t, 7.2Hz), 1.61(9H, s) 2.57(2H, bg, 7.2Hz), 4.52-4.84(1H, in), 6.43- 6.64(1H, mn), 6.64-7.02(2H, mn), 7.12-7.48(2H, in), 7.48-7.67(lH, in).

0 Exaple133 Preparation of cycloartenyl ester of 3-iethoxy-4-valeryloxy-a-propylcinnaic acid The title compound was prepared according to following the procedure of Example 119 but 3-inethoxywas used as a starting material, in place of 4-butyryloxy 203-inethoxy-ct-ethylcinlaiic acid. The yield was 23.2 g (77.6 r.p. 113 114'C.

Specific rotation [a±2 34.2- (C 1.00, CHCl) Analysis Calcd. for C 48

H

72 0 5 (M.W.729.06): C,79.07; H 9.95. Found: C,79.13; H 9.88.

Example 134 Preparation of cycloartenyl ester of 4-hydroxy-3-inethoxy-a-propylcinnanic acid 18 1 The title compound was prepared according to following the procedure of Example 120 but cycloartenyl ester of 3-methoxy-4-valeryloxy-a-propyl-cinnamic acid (23.3 g, 0.032 mole) obtained according to the procedure of Example 133 was used as a starting material. The yield was 18.1 g (87.6 m.p. 122 1230C.

20 Specific rotation [a]D 41.20(C 1.00, CHCl1) Analysis Calcd. for C 43

H

64 0 4 (M.W.644.94): C,90.07; H,10.00. Found: C,0.14; H,9.97.

Example 135 Preparation of cycloartenyl ester of 4-capryloxy-3-methoxy-a-butylcinnamic acid a The title compound was prepared according to following the procedure of Example 119 but 4-capryloxy- 15 3-methoxy-a-butylcinnamic acid (19.2 g, 0.0551 mole) Irtu was used in place of 4-butyryloxy-3-methoxy-a-ethylcinnamic acid. The yield was 22.7 g(54.4 m.p. 100 1010C.

20 Specific rotation 2 33.50(C 1.00, CHCl) Analysis Calcd. for C 50

H

76 0 5 (M.W.757.11): C,79.31; H,10.12. Found: C,79.38; H,10.05.

Example 136 Preparation of cycloartenyl ester of 4-hydroxy- 3-methoxy-a-butylcinnamic acid I I t The title compound was prepared according to following the procedure of Example 120 but cycroartenyl ester of 4-capryloxy-3-methoxy-a-butylcinnamic acid

L

188 1 (24.4 g, 0.032 mole) was used as a starting material.

The yield was 17.8 g (84.4 m.p. 110 111iC.

20 Specific rotation [a]D 40.60(C 1.00, CHC1) Analysis Calcd. for C44H 604(M.W.672.99): C,80.19; H,10.10. Found: C,80.24; H,10.05.

Example 137 Preparation of cyclobranyl ester of 4hydroxy-3-methoxy-a-butylcinnamic acid The title compound was prepared according to following the procedure of Example 120 but cyclobranyl ester of 4-capryloxy-3-methoxy-a-butylcinnamic acid (26.2 g, 0.034 mole) was used as a starting material.

The yield was 18.4 g ((80.4 m.p. 132 1330C.

20 Specific rotation [alD 37.00(C 1.00, CHC3) Analysis Calcd. for C 45

H

68 0 4 (M.W.672.99): C,80.31; H,10.18. Found: C,80.39; H,10.04.

Example 138 Preparation of 24-methylenecycloartanyl ester of 4-hydroxy-3-methoxy-a-butylcinnamic acid The title compound was prepared according to following the procedure of Example 120 but 24-methylenecycloartanyl ester of 4-capryloxy-3-methoxy-a-butylcinnamic acid (26.2 g, 0.034 mole) was used as a starting material. The yield was 18.1 g (79.1 m.p. 124 1250C.

20 Specific rotation [a]D 39.8C 1.00, CHC13 1 Analysis Calcd. for CH 0MW629):C8.1 680 4 C831 H,10.18. Found: C,80.25; H,10.22.

Example 139 Preparation of cycloartenyl ester of 3ethoxy-4-propionyloxy-a-methylcinnamic acid The titel compound was prepared according to following the procedure of Example 120 but cycloartenyl ester of 3 -ethoxy-4-.propionyloxy-ca-methylcinnamic acid (22.0 g, 0.032 mole) was used as a starting material.

The yield was 15.8 g (78.2 m.p. 132 133'C.

Speifi roatin [xl20 43.90 (C 1.00, C~ Spcfcrtain[]D Hl 3 Analysis Calcd. for C 42

H

62 0 4 (M.W.630.92): C,79.95; H,9.91. Found: C,79.90; H 9.99.

Example 140 Preparation of cyclobranyl ester of 3ethoxy-4 -hydroxy-a-methylcinnamic acid The title compound was prepared according to following the procedure of Example 120 but cyclobranyl ester of 3-ethoxy-4-propionyloxy-.a-methylcinnamic acid (21.7 g, 0.031 mole) was used as a starting material.

The yield was 16.1 g (80.5 174 1751C.

Specific rotation [a]D 42.4-(C 1.00, CHCl) Analysis Calcd. for C 43

H

64 0 4 (.W.644.94): C,80.07; H,10.00. Found: C,80.18; H,10.05.

Example 141 Preparation of 24-methylenecycloartanyl ester of 3-ethoxy-4--hydroxy-a-methylcinnamic acid 19 0 1 The title compound was prepared according to following the procedure of Example 120 but 24-methylenecycloartanyl ester of 3-ethoxy-4-propionyloxy-a-methylcinnamic acid (22.2 g, 0.0317 mole) was used as a starting material. The yield was 16.7 g (81.6 m.p. 134 135 0

C.

20 Specific rotation [a]D 40.20(C 1.00, CHC1 3 Analysis Calcd. for C43H6404(M.W.644.94): C,80.07; H,10.00. Found: C,80.13; H,9.92.

Example 142 Preparation of cycloartenyl ester of 3ethoxy-4-hydroxy-a-ethylcinnamic acid o' The title compound was prepared according to following the procedure of Example 120 but cycloartenyl ester of 4-butyryloxy-3-ethoxy-a-methylcinnamic acid (21.5 g, 0.030 mole) was used as a starting material.

The yield was 15.4 g (79.6 m.p. 124 125 0

C.

41 (C 1.00 Specific rotation [a]D 41.2°(C 1.00, CHC1 3 Analysis Calcd. for C 43

H

64 0 4 (M.W.644.94): C,80.07; H,10.00. Found: C,80.04; H,10.08.

Example 143 Preparation of cycloartenyl ester of 3ethoxy-4-hydroxy-a-propylcinnamic acid The title compound was prepared according to 4 e 25 following the procedure of Example 120 but cycloartenyl ester of 3-ethoxy-4-valeryloxy-a-propylcinnamic acid (26.0 g, 0.035 mole) was used as a starting material.

191 1 The yield was 16.8 g (72.8 m.p. 111 1120C.

Specific rotation [a]20 40.7 0 (C 1.00, CHC1 Analysis Calcd. for C44H6604 (M.W.658.97): C,80.19; H,10.10. Found: C,80.26; H,10.02.

Example 144 Preparation of cyclobranyl ester of 3ethoxy-4-hydroxy-a-propylcinnamic acid The title compound was prepared according to follwoing the procedure of Example 120 but cyclobranyl ester of 3-ethoxy-4-valeryloxy-a-propylcinnamic acid (24.2 g, 0.032 mole) was used as a starting material.

The yield was 16.7 g(72.8 m.p. 134 135 0

C.

20 Specific rotation [a]D 37.10(C 1.00, CHC1) SAnalysis Calcd. for C H 04 (M.W.672.99): C,80.31; i. o a45 68 4 15 H,10.18. Found: C,80.25; H,10.24.

0 0 Example 145 Preparation of cycloartenyl ester of 3ethoxy-4-hydroxy-a-butylcinnamic acid 00 "The title compound was prepared according to following the procedure of Example 120 but cycloartenyl ester of 4-capryloxy-3-ethoxy-a-butylcinnamic acid (23.1 g, 0.030 mole) was used as a starting material.

The yield was 16.2 g (80.2 m.p. 99 1000C.

20 Specific rotation 40.00(C 1.00, CHC13) Analysis Calcd. for C45H6804(M.W.672.99); C,80.31; H,10.18. Found: C,80.21; H,10.22.

I-I

19' 1 Example 146 Preparation of cycloartenyl ester of 4hydroxy-3-propoxy-a-methylcinnamic acid The title compound was prepared according to following the procedure of Example 120 but cycloartenyl ester of 4-propionyloxy-3-propoxy-a-methylcinnamic acid (23.1 g, 0.033 mole) was used as a starting material.

The yield was 17.2 g (80.8 m.p. 138 139 0

C.

43 (C 1.00 Specific rotation [a]D 43.70(C 1.00, CHC1 3 Analysis Calcd. for C43 H 0(M.W.644.94): C,80.07; H,10.00. Found: C,80.19; H,10.04.

Example 147 Preparation of cycloartenyl ester of 4hydroxy-3-butoxy-a-methylcinnamic acid The title compound was prepared according to following the procedure of Example 120 but cycloartenyl ester of 4-propionyloxy-3-butoxy-a-methylcinnamic acid (22.9 g, 0.032 mole) was used as a starting material.

The yield was 16.5 g (78.2 m.p. 126 127 0

C.

SSpecific rotation [a2 0 39.70(C 1.00, CHC1 3 Analysis Calcd. for C 44

H

66 0 4 (M.W.658.97): C,80.19; H,10.10. Found: C,80.24; H,10.03.

Example 148 Preparation of 24-methylenecycloartanyl ester of 4-butyryloxy-3-methoxy-a-ethylcinnamic acid The title compound was prepared according to follwoing the procedure of Example 121 but 24-methyleneli I I I t t 1cycloartanol (18.1 g, 0.041 mole) was used as a starting material. The yield was 22.8 g (77.8 127 128 0

C.

Specific rotation [a 20 35.10 (C 1.00, CHCl) Analysis Calcd. for C 47

H

70 0 5 (M.W.715.03): C,78.94; H,9.87. Found: C,78.90; H,9.79.

Example 149 Preparation of 24-methylenecycloartanyl ester of 4-hydroxy-3-methoxy-a-ethylcinnamic acid The title compound was prepared according to following the procedure of Example 122 but 24-methylenecycloartanyl ester of 4-butyryloxy-3-methoxy-a-ethylcinnamic acid (21.5 g, 0.0301 mole) was used as a starting material. The yield was 17.3 g (89.J- m.p. 137 138 0

C.

Specific rotation [ac]D2 40.7 0 (C 1.00, CHCl) Analysis, for C 43

H

64 0 4 (M.W.644.94) Calcd. 80.07, H 10.00 Found C 80.11, H 9.93 Example 150 Preparation of cyclobranyl ester of 4hydroxy-3-piropoxy-a-ethylcinnamic acid The title compound was prepared according to following the procedure of Example 120 where cyclobranyl ester of 4-butyryloxy-3-propoxy-a--ethylcinnamic acid (22.1 g, 0.0297 mole) was used in place of cycloartenyl ester of 4-butyryloxy-3-metihoxy-ca-ethylcinnamic acid.

CL

1 i 19-1 1 The yield was 17.8 g (89.1 m.p. 140 141 0

C.

Specific rotation [a 20 36.80(C 1.00, CHC1 Analysis. for C 45 H6 8 0 4 (M.W.672.99) Calcd. C 80.31, H 10.18 Found C 80.36, H 10.12 Example 151 Preparation of 24-methylenecycloartanyl ester of 4-hydroxy-3-propoxy-a-propylcinnamic acid The title compound was prepared according to following the procedure of Example 120 where 24-methylenecycloartanyl ester of 3-propoxy-4-valeryloxy-a-propylcinnamic acid (23.5 g, 0.0305 mole) was used in place of cycloartenyl ester of 4-butyryloxy-3-methoxy-aethylcinnamic acid. The yield was 17.2 g (82.1 121 0

C.

20 Specific rotation [a]20 39.10(C 1.00, CHC1) Analysis, for C 46

H

70 0 4 (M.W.687.02) Calcd. C 80.41, H 10.27 Found C 80.32, H 10.34 Examples 152 154 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of 3-propionyloxy-a-methylcinnamic acid The title compounds were prepared according to following the procedure of Example 123 where cycloartenol (21.3 g, 0.050 mole), cyclobranol (22.0 g, 0.050 mole),

I

4 1 1 11111~- 195 1 and 24-methylenecycloartanol (22.0 g, 0.050 mole), respectively, and 3-propionyloxy-a--methyl-cinnamic acid (17.6 g, 0.075 mole) were used for each preparation.

The yield m.p. and specific rotation 20 {[a2D (C 1.00, CHC13)}of each product were as follows: [allD 00 0o 0 00 4 0 S It 0 0't0 FI14 Example Triterpenyl ester of Yield Melting Specific No. 3-propionyloxy-a- point rotation methylcinnamic acid (oC) [al] 152 Cycloartenyl ester 83.1 80 81 +44.50 153 Cyclobranyl ester 82.4 100-101 +34.30 154 24-Methylene 82.0 85 86 +44.00 cycloartanyl ester Examples 155 157 Preparation of cycloartenyl, cyclobranyl, and 24-methylene cycloartanyl ester of 3-hydroxya-methylcinnamic acid The title compounds were prepared according to 20 following the procedure of Example 124 where the compounds (each 0.042 mole) of Examples 152 154 were used respectively. The yield and specific 20 rotation (C 1.00, CHCI 3 of each produce were as follows: 1 119G I *I ttti ti I te Example Triterpenyl ester of Yield Melting Specific No. 3-hydroxy-a-methyl- point rotation cinnamic acid [a]D 155 Cycloartenyl ester 86.2 178-179 +44.30 156 -Cyclobranyl ester 87.2 191-192 +45.20 24-Methylene- 157 85.8 188-189 +42.70 cycloartanyl ester Examples 158 160 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl ester of 3-butyryloxy-a-ethylcinnamic acid The title compounds were prepared according to 15 following the procedure of Example 129 where 3-butyryloxya-ethylcinnamic acid (3.50 g, 0.0135 mole) and cycloartenol (2.85 g, 0.0067 mole), cyclobranol (2.95g, 0.0067 mole), and 24-methylenecycloartanol (2.95 g, 0.0067 mole), respectively were used for each preparation. The yield m.p. and specific rotation {[aD (C 1.00, CHC13)} of each product '7ere as follows: I~ 19 7 Example Triterpenyl ester of Yield Melting Specific No. 3-butyryloxy-a- point rotation ethylcinnamic acid C) [a]

D

158 Cycloartenyl ester 81.4 82 83 +41.40 159 Cyclobranyl ester 82.3 112-113 +37.40 24-Methylene 160 79.4 89 90 +40.80 cycloartanyl ester 00 0 0 0 V o o 00 00 a o oo ae 0 a 4000 u oao 0 *0 too O 0 9 090 0 Examples 161 163 Preparation of cycloartenol, cyclobranol, and 24-methylenecycloartanol ester of 3-hydroxy-a-ethylcinnamic acid The title compounds were prepared according to 15 following the porcedure of Example 129 where the compounds (each 0.003 mole) of Examples 158 160 were used, respectively. The yield m.p. (oC, and 20 specific rotation {[a]20 (C 1.00, CHC1 3 of each product were as follows: Example Triterpenyl ester of Yield Melting Specific No. 3-hydroxy-a-ethyl- point rotation cinnamic acid (Co) ]20

D

161 Cycloartenyl ester 92.4 161-162 +44.80 162 Cyclobranyl ester 93.4 188-189 +46.00 24-Methylene- 163 91.8 173-174 +44.20 cycloartanyl ester 0 Otto 0 0 'sot 09 09 90 0C 0 198 1 Examples 164 165 Preparation of cycloartenyl and cyclobranyl esters of 2-hydroxy-a-methylcinnamic acid Cycloartenyl and cyclobranyl esters of 2propionyloxv-a-methylcinnamic acid (27.5 g, yield 85.5 27.5 g, yield 82.8 respectively) were prepared according to following the procedure of Example 123 where cycloartenol (21.3 g, 0.050 mole) and cyclobranol (22.0 g, 0.050 mole), respectively, and 2-propionyloxy-a-methylcinnamic acid (17.6 g, 0.075 mole were used) for preparation. Using these esters

S

t (each 24.5 the title compounds were prepared according to following the procedure of Example 124.

The yield m.p. and specific rotation 20 (C 1.00, CHC13)} of each product were as follows: D5{[ F I* 8t I Example Triterpenyl ester of Yield Melting Specific No. 2-hydroxy-a- point rotation methylcinnamic acid (oC) [a]0

D

164 Cycloartenyl ester 85.4 185-186 +46.40 165 Cyclobranyl ester 86.2 197-198 +48.70 Example 166 Preparation of cycloartenyl ester c: 3-methoxy-4-nitrobenzoic acid To 3-methoxy-4-nitrobenzoic acid (15.0 g, 0.076 mole) were added thionyl chloride (34 mt, 6 equivalents) 'p 199 1 and dimethylformamide (0.5 m) and the mixture was stirred at 600C for 2 hours. Then the reaction mixture was concentrated under reduced pressure and the residue was mixed with dioxane (75 mf) at 0°C.

Thereto was added a solution of cycloartenol (25.0 g, 0.059 mole) in pyridine (110 mZ). This reaction mixture was stirred at 700C for 20 minutes. After thus completing the reaction, the solvents were removed by distillation under reduced pressure. The resulting residue was dissolved in chloroform, and the chloroform layer was washed with saturated aqueous NaHCO 3 solution, and dried. The chloroform solution was concentrated under reduced pressure. The residual crystals were recrystallized from methylene chloride-methanol giving cycloartenyl ester of 3-methoxy- 4-nitrobenzoic acid (30.5 g) in a 85.3 yield.

m.p. 182 1830C.

22.5 Specific rotation 22 57.7(C 1.00, CHC13 Analysis, for C38H55NO 5 (M.W.605.82) 20 Cald. C 75.33, H 9.15, N 2.31 Found C 75.42, H 9.07, N 2.36 -1 IRv, KBr(dm 2940, 1720, 1610, 1530, 1410, 1350, 1310, 1290, 1245.

PMR(CDC1 3 0.38(1H, 1/2ABq, 4.2Hz), 0.62(1H, 1/2ABq, 4.2Hz), 0.50-2.36(27H, 0.95(H1 0.97 (3H, 1.04(3H, 1.60(3H, 1.69(3H, s), 4.00(3H, 4.50-5.32(2H, 7.42-8.01(3H, m).

r Lr t 9 4 g I 2~

I

200 1 Example 167 Preparation of cycloartenyl ester of 4amino-3-methoxy benzoic acid Acetic acid (400 mi) and dioxane (400 m) were added to cycloartenyl ester of 3-methoxy-4-nitrobenzoic acid (40.0 g, 0.066 mole) prepared according to the procedure of -Example 166. Thereto 6N-HCZ-dioxane (22 mt, 2 equivalents) and zinc powder (40 g) were added at 0°C and the mixture was stirred at 25°C for 2 hours.

After the reaction, zinc powder was removed by filtration.

The filtrate was concentrated under reduced pressure, and the residue was extracted with chloroform. The chloroform extracts were washed successively with water and saturated aqueous NaHCO solution, then dried, and o a concentrated. The residual crystals were recrystallized S 15 from methylene chloride-methanol giving cycloartenyl ester 4-amino-3-methoxybenzoic acid (32 g) in a 84.1 yield. m.p. 186 188 0

C.

26.5 Specific rotation [a]D 64.3(C 1.00, CHC3) Analysis, for C 3H57NO (M.W.575.83) Calcd. C 79.26, H 9.98, N 2.43 Found C 79.32, H 9.99, N 2.39 IRV, KBr(cm-): 3450, 3350, 2930, 1700, 1620, 1520, 1460, 1305, 1285, 1260, 1220, 1180, 1105.

PMR(CDC1 6: 0.36(1H, 1/2ABq, 4.2Hz), 0.61(1H, 1/2ABq, 4.2Hz), 0.48-2.39(27H, 1.61(3H, 1.67 S(3H, 3.88(3H, 4.20(2H, bs), 4.51-5.31 (2H, 6.46-6.77(1H, 7.30-7.71(2H, m).

201 1 Example 168 Preparation of cyclobranyl ester of 3methoxy-4-nitrobenzoic acid 3-Methoxy-4-nitrobenzoic acid (50.0 g, 0.254 mole) was allowed to react with thionyl chloride (60 m9, 3.2 equivalents) by adding dimethylformamide mf) and the mixture was stirred at 60 0 C for 2 hours. Then the reaction mixture was concentrated under reduced pressure. Dioxane (100 mZ) was added thereto, and further a solution of cyclobranol (93 g, 0.211 mole) in pyridine (150 mk) was added at 0°C. The 1mixture was stirred at 70 0 C for 30 minutes. After the reaction, the solvents were removed by distillation under reduced pressure. The resulting residue was dissolved in chloroform,and the chloroform layer was washed with saturated aqueous NaHCO 3 solution, and dried. This chloroform solution was concentrated under reduced pressure, and the residual crystals were recrystallized from chloroform-ethanol v/v), giving cyclobranyl ester of 3-methoxy-4-nitrobenzoic acid (94.4 g) in a 72.1 yield, m.p. 213 214 0

C.

7 I 25.5 t Specific rotation [a]D5 53.9 0 (C 1.00, CHC Analysis, for C39H NO5(M.W.619.85) Calcd. C 75.57, H 9.27, N 2.26 Sr 1 Found C 75.63, H 9.22, N 2.33 -1 IRv, KBr(cm1): 2930, 1715, 1610, 1530, 1410, 1360, 1310, 1285, 1240.

PMR(CDC1 3 0.39(1H, 1/2ABq, 4.8Hz), 0.62(1H, 1/2ABq, S 20.2 1 4.8Hz), 0.50-2.28(27H, 0.92(6H, 0.99 (3H, 1.05(3H, 1.63(9H, 4.01(3H, s), 4.62-5.03(lH, 7.48-7.96(3H, m).

Example 169 Preparation of cyclobranyl ester of 4-amino- 3-methoxybenzoic acid Cyclobranyl ester of 3-methoxy-4-nitrobenzoic acid (94.3 g, 0.152 mole) prepared according to the procedure of Example 168 was suspended in a mixture of acetic acid (1.2 p) and tetrahydrofuran (1.2 and thereto 6N-HCZ-dioxane (100 mf) and zinc powder (94 g) were added, and the reaction mixture was stirred at 0 C for 2 hours. After the reaction, zinc powder was removed by filtration. The filtrate was concentrated under reduced pressure and the residue was extracted with chloroform. The chloroform extracts were washed successively with water and saturated aqueous NaHCO 3 solution, then dried, and concentrated. The residual crystals were recrystallized from chloroform-ethanol giving cyclobranyl ester of 4-amino-3methoxybenzoic acid ester (64.2 g) in a 71.5 yield.

m.p. 235-236 0

C.

t 2 Specific rotation 60.8 0 (C 1.00, CHC1) D 3 t Analysis, for C39H59NO(M.W.589.86) Calcd. C 79.41, H 10.08, N 2.37 Found C 79.49, H 10.12, N 2.42 IR, KBr(cm 3450, 350 2900, 1680 1620 1310, IRv, KBr(cm 3450, 3350, 2900, 1680, 1620, 1310, I -3 1 1280, 1260, 1110.

PMR(CDC 3 0.36(lH, 1/2ABq, 4.8Hz), 0.61(lH, 1/2ABq, 4.8Hz), 0.50-2.20(27H, 0.89(6H, 0.96 (3H, 1.01(3H, 3.85(3H, 3.9204.36 (2H, bs), 4.51-4.91(lH, 6.42-6.72(1H, m), 7.26-7.72(2H, m).

Example 170 Preparation of cycloartenyl ester of zcid 2-methoxy-5-nitrobenzoic acid (17,3 g, 0.088 mole) was allowed to react thionyl chloride (65 mt, 10 equivalents) by adding dimethylformamide (0.3 m,) P" and the mixture was stirred at 50 0 C for 1.5 hours. Then, the reaction mixture was concentrated under reduced pressure, dioxane (125 mk) was added thereto and further a solution of cycloartenol (25.0 g, 0.059 mole) in pyridine (125 mk) was added dropwise at 0°C. This reaction mixture was stirred at 60 0 C for 1.5 hours.

After the reaction, the solvents were removed by distillation under reduced pressure, the residue was extracted with chloroform, and the chloroform extracts were washed successively with water and saturated aqueous NaHCO 3 solution, then dried, and concentrated.

The residue was recrystallized from methylene chloride-hexane giving cycloartenyl ester of 2-methoxy-5-nitrobenzoic acid (31.5 g) in an 88.7 yield, m.p. 186 1870C.

r -t, 3-;ii OQ9* 0 t 204 25 1 Specific rotation [a]D 43.9(C 1.00, CHC13 Analysis, for C38H55NO 5 (M.W.605.82) Calcd. C 75.33, H 9.15, N 2.31 Found C 75.30, H 9.22, N 2.29 IRv, KBr(cm- 1 2930, 1695, 1610, 1520, 1340, 1280, 1135.

PMR(CDC1 3 0.39(1H, l/2ABq, 4.2 Hz), 0.62(1H, 1/2ABq, 4.2 Hz), 0.50-2.40(27H, 0.90(3H, 0.96 (6H, 1.01(3H, 2.60(3H, bs), 2.68(3H, bs), 4.00(3H, 4.65-5.30(3H, 7.08(1H, d, 9.4Hz) 8.34(1H, dd, 3.0Hz, 9.4Hz), 8.64(1H, d, Example 171 Preparation of cycloartenyl ester of amino-2-methoxybenzoic acid Cycloartenyl ester of acid (34.0 g, 0.056 mole) prepared according to the procedure of Example 170 was suspended in acetic acid (1.2 f) at 20 0 C, and thereto 6N-HC1-dioxane (19 mt, 2 equivalent) and zinc powder (68 g) were added. The mixture was stirred at 30 0 C for 1 hour. Then, zinc powder was removed by filtration, and the filtrate was concentrated under reduced pressure and the residue was extracted with chloroform. The chloroform extracts were washed successively with water and saturated aqueous NaHCO 3 solution, then dried, concentrated, and the residual crystals were recrystallized from methylene chloride-hexane giving "2P& c 205 1 cycloartenyl ester of 5-amino-2-methoxybenzoic acid (27.2 g) in a 84.4 yield. m.p. 180 182 0

C.

26.5 Specific rotation []265 47.80(C 1.00, CHCl Analysis, for C38H57NO 3 (M.W.575.83) Calcd. C 79.26, H 9.98, N 2.43 Found C 79.32, H 9.94, N 2.41 IRv, KBr(cm-): 3450, 3350, 2900, 2860, 1690, 1630, 1500, 1440, 1300, 1270, 1245.

PMR(CDC13)6: 0.38(1H, 1/2ABq, 4.2Hz), 0.59(1H, 1/2ABq, 4.2Hz), 0.50-2.30(27H, m) 0.90(6H, 0.93 (6H, 1.59(3H, bs), 1.67(3H, bs), 3.55(2H, bs), 3.88(3H, 4.50-5.30(2H, 6.68-7.24(3H, m).

Example 172 Preparation of cyclobranyl ester of 2methoxy-5-nitrobenzoic acid acid (11.6 g, 0.059 mole) was allowed to react with thionyl chloride (20 m) and dimethylformamide (0.2 mz) and the mixture was stirred at 50 0 C for 2 hours. Then the reaction mixture was concentrated under reduced pressure. Toluene (150 mi), pyridine (30 mt), and further cyclobranol (20 g, 0.045 mole) were added to the residue and the mixture was stirred at 60 0 C for 2 hours. After the reaction, S' the solvents were removed by distillation under reduced pressure and the residue was extracted with chloroform.

The chloroform extracts were washed successively with water and saturated aqueous NaHCO 3 solution, then dried, 1 concentrated, and the residue was recrystallized from chloroform-ethanol v/v),giving cyclobranyl ester of 2-methoxy-5-nitrobenzoic acid (25.9 g) in a 92.0 yield. m.p. 207 208 0

C.

25 Specific rotation [a]D 32.5 0 (C 1.00, CHC Analysis, for C H 5NO M W 619 85 Calcd. C 75.57, H 9.27, N 2.26 4.8Hz), 0.76-2.24(27H, 0.91(3H, 0.96 (6H, 1.01(3H, 1.63(9H, 4.01(3H, s), 4.64-5.02(lH, 7.06(1H, d, 9.6Hz), 8.34(1H, dd, 9.6Hz, 3.6Hz), 8.67(1 H, d, 3.6Hz).

Example 173 Preparation of cyclobranyl ester of 2-methoxybenzoic acid Cyclobranyl ester of acid (25.0 g, 0.040 mole) prepared according to the procedure of Example 172 was suspended in acetic acid (1 and thereto 6N-HCl-dioxane (21 m) and zinc S powder (25.0 g) were added. The mixture was stirred at 30 0 C for 2 hours. After the reaction, zinc powder was removed by filtration. The filtrate was concentrated under reduced pressure, and the residue was extracted with chloroform. The extracts were washed successively 1

VY

1 with water and saturated aqueous NaHCO 3 solution, then dried, concentrated, and the residue was recrystallized from chloroform-ethanol v/v) giving cyclobranyl ester of 5-amino-2-methoxybenzoic acid (13.7 g) in a 57.5 yield. m.P. 193 1951C.

Specific rotation [at] 26.5 41.50 (C 1.00, CHCl 3 Analysis, for C H NO(.5886 39 59 0 3 (..886 Calcd. C 79.41, H 10.08, N 2.37 Found C 79.35, H 10.15, N 2.35 IRv, KBr(cm 3430, 3350, 2930, 1690, 1500, 1460, 1430, 1310, 1270, 1245.

PMRCD3&)6 0.36(lH, l/2ABq, 4.8Hz), 0.61(lH, l/2ABq, 4.8Hz), 0.50-2.28(27H, mn), 0.92(3H, 0.96 (6H, 1.00(3H, 1.64(9H, 2.88-3.26 (2H, mn), 3.81(3H, 4.52-5.02(lH, mn), 6.74- 6.90(lH, in), 7.08-7.22(2H, in).

Examples 174 1.76 Preparation of cyclc-artenyl, cyclobranyl, and 24-metnylenecycloartanyl esters of 3-methoxy-4-nitrocinnamic acid The title compounds were prepared according to following the procedure of Example 166 where 3-methoxy-4nitrocinnamic acid (17.0 g, 0.076 mole) for each preparation and cycloartenol (25.0 cyclobranol (26.0 and 24- 4 25 methylenecycloartanol (26.0 respectively were used.

The yield m.p. and specific rotation {[a]D2 (C 1.00, CHCl 3 of each product were as follows: r l f, o u 1 I ~h, oao s: too r, 0 0 a ooooo 0 0 09 0 0 9o a a a• 0 0 0 9.99 9 9000 0 C E 00L t e t 9 I 9 I I 9 1 90t 9990« Example Triterpenyl ester of Yield Melting Specific No. 3-methoxy-4-nitro- point rotation cinnamic acid [a]

D

174 Cycloartenyl ester 86.2 190-194 +43.60 175 Cyclobranyl ester 85.3 220-221 +42.30 24-Methylene- 176 85.8 210-211 +43.00 cycloartanyl ester Examples 177 179 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl ester of 4-amino-3-methoxycinnamic acid The title compounds were prepared according to 15 following the procedure of Example 167 but where using cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of 3-methoxy-4-nitrocinnamic acid (41.7 g, 42.6 g and 42.6 g, respectively, each 0.066 mole) obtained in Examples 174- 176 were used, respectively. The yield 25 20 m.p. and specific rotation (C 1.00, CHC1 3 of each product were as follows: 20W9 00 00 0 000 '*0 0 0 6 0 o 0000 0.00 0 00 *04 000 00 00 490 Example Triterpenyl ester of Yield Melting Specific No. 4-amino-3-methoxy- W% point rotation cinnamic acid (OC) (]2 177 Cycloartenyl ester 85.2 194-195 +42.30 178 Cyclobranyl ester 85.8 240-241 +41.00 2 4-Methylene- 179 86.4 227-228 +41.80 cycloartanyl ester Examples 180 182 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of acid The title compounds were prepa.,ed according to 15 following the procedure of Example 170 where 2-ethoxy- 5-nitrocinnamic acid (19.5 g, 0.082 mole) for each preparation and cycloartenol (25.0 g, 0.059 mole), cyclobranol (26.0 g, 0.059 mole) and 24-methylenecycloartanol (26.0 g, 0.059 mole) respectively were used.

The yield m.p. and specific rotation (C 1.00, CHCl of each product were as follows: r 7 2.1 21{ i1i I ~i l~ir t I I 1I I t If f I I

SC

o *i Example Triterpenyl ester of Yield Melting Specific No. 2-ethoxy-5-nitro- point rotation cinnamic acid [a]

D

180 Cycloartenyl ester 87.8 182-183 +42.70 181 Cyclobranyl ester 88.4 203-204 +40.40 24-Methylene- 182 87.2 198-199 +42.10 cycloartanyl ester Examples 183 185 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of 5-amino-2-ethoxycinnamic acid The title compounds were prepared according to 15 following the procedure of Example 171 where cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of 2-ethoxy-5-nitrocinnamic acid (36.2 g, 37.0 g, and 37.0 g, respectively, each 0.056 mole) obtained in Examples 180 182, respectively were used. The yield 20 m.p. and specific rotation 25 (C 1.00, CHCl 3 of each product were as follows: (r 211.

1 IiI t* t C I I L4 4

-I

Example Triterpenyl ester of Yield Melting Specific No. 5-amino-2-ethoxy- point rotation cinnamic acid

D

183 Cycloartenyl ester 85.3 176-177 +43.20 184 Cyclobranyl ester 84.8 190-191 +41.00 24-Methylene- 185 85.8 181-182 +42.20 cycloartanyl ester Example 186 188 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl ester of 3-methoxy-4-nitro-a-methylcinnamic acid The title compounds were prepared according to 15 following the procedure of Example 166 where 3-methoxy- 4-nitro-a-methylcinnamic acid (17.3 g, 0.073 mole) for each preparation and cycloartenol (25.0 g, 0.059 mole), cyclobranol (26.0 g, 0.059 mole), and 24-methylenecycloartanol (26.0 g, 0.059 mole), respectively were used. The yield m.p. and specific rotation (C 1.00, CHC1 3 of each product were as follows:

*A

i' i 212 Example Triterpenyl ester of Yield Melting Specific No. 3-methoxy-4-nitro-a- point rotation methylcinnamic acid (oC) 2a] 186 Cycloartenyl ester 83.4 178-180 +44.30 187 Cyclobranyl ester 84.2 208-209 +43.70 24-Methylene- 188 82.5 199-200 +44.10 cycloartanyl ester 0000 0 Q0 00 00f o ao 0 0 a o 9o 00 o o *000 0 0000 00 e a e oo 00a 0 0 0*6 0 a OI 0 0 0 0 e 9 0 Examples 189 191 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of 4-amino-3-methoxy-a-methylcinnamic acid The title compounds were prepared according to 15 following the procedure of Example 167 where cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of 3-methoxy-4-nitro-a-methylcinnamic acid (43.6 g, 44.5 g, and 44.5 g, respectively, each 0.066 mole) obtained in Examples 186 188, respectively 20 were. used. The yield m.p. and specific 25 rotation (C 1.00, CHC1 3 of each product were as follows: .il i i irB L i Si i::L i c 213 Example Triterpenyl ester of Yield Melting Specific No. 4-amino-3-methoxy-a-- point rotation methylcinnamic acid [a] D 189 Cycloartenyl ester 85.6 183-184 +43.00 190 Cyclobranyl ester 84.5 225-226 +42.00 24-Methylene- 191 85.0 213-214 +42.70 cycloartanyl ester Examples 192 194 Preparation of cycloartenyl cyclobranyl, and 24-methylenecycloartanyl esters of 5-nitro-2-propoxy-a-methylcinnamic acid The title compounds were prepared according to following the procedure of Example 170 where 5-nitro-2propoxy-a-methylcinnamic acid (21.2 g, 0.080 mole) for each preparation and cycloartenol (25.0 g, 0.059 mole), cyclobranol (26.0 g, 0.059 mole), and 24-methylenecycloartanol (26.0 g, 0.059 mole), respectively were used. The yield m.p. and specific rotation {[alD (C 1.00, CHC1 3 of each product were as follows: 4C 4t r i -ii i; i cc 214 Example Triterpenyl ester of Yield Melting Specific No. 5-nitro-2-propoxy-a- point rotation methylcinnamic acid []D 192 Cycloartenyl ester 86.4 184-185 +43.20 193 Cyclobranyl ester 88,4 204-205 +41.30 24-Methylene 194 87.2 196-197 +42.70 cycloartanyl ester rr t t 4 i 44 *#4

I

IC

SIt I o rI C C C I I Examples 195 197 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of 5-amino-2-propoxy-a-methylcinnamic acid The title compounds were prepared according to 15 f6llowing the procedure of Example 171 where cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of 5-nitro-2-propoxy-a-methylcinnamic acid (37.7 g, 38.5 g, and 38.5g respectively, each 0.056 mole) obtained in Examples 192 194, respectively, were used. The yield m.p. and specific 25 rotation {[a2D (C 1.00, CHC1 3 of each product were as follows: 1 ccl UI-- 211) Example Triterpenyl ester of Yield Melting Specific No. 5-amino-2-propoxy-a- point rotation methylcinnamic acid [al

D

195 Cycloartenyl ester 84.4 175-176 +44.20 196 Cyclobranyl ester 83.5 194-195 +41.70 24-Methylene- 197 84.2 187-188 +43.60 cycloartanyl ester o 6 46r *6 *4 6 6L(1 6e4 46 4 I 6 tt 1* 4* 4I' 44 C C 4 Examples 198 and 199 Preparation of cycloartenyl and cyclobranyl esters of 3-methoxy-4-nitro-aisopropyl cinnamic acid The title compounds were prepared according to 15 following the procedure of Example 166 where 3-methoxy 4 -nitro-a-isopropylcinnamic acid (19.1 g, 0.072 mole) for each preparation and cycloartenol (25.0 g, 0.059 mole) and cyclobranol (26.0 g, 0.059 mole), respectively were used. The yield m.p. and specific 25 20 rotation (C 1.00, CHC1 3 of each product were as follows: 21 Example Triterpenyl ester of Yield melting Specific No. 3-methoxy-4-nitro-a- M% point rotation isopropylcinnamic (OC) acid 198 Cycloartenyl ester 84.2 155-156 +41.20 199 Cuclobranyl ester 184.6 1190-191 +40.50 Is', *1 ,i' a 0

I

I

~**sII a

II

S

a S ItS 4 *54 a t it C C C C

.C

St C S C CC S CCC C

S

P t tC C~ C C C C C Examples 200 and 201 Preparation of cycloartenyl and cyclobranyl esters of 4-amino-3-methoxy-aisopropylcinnamic acid The title compounds were prepared according to following the procedure of Example 167 where cycloartenyl 15 and cyclobranyl esters of 3-methoxy-4-nitro-a-isopropylcinnamic acid (44.5 g and 45.4 g, respectively, each 0.066 mole) obtained in Examples 198 and 199, respectively were used. The yield m.p. and specific rotation 2 (C 1.00, CHCl of each product were as 20 f Qllows: Example Triterpenyl ester of Yield Melting Specific No. 4-amino-3-methoxy-a- M% point rotation isopropylcinnamic acid 0 C) ___[ciD 25 200 Cycloartenyl ester 85.8 163-164 +-40.80 201 -Cyclobranyl ester 84.2 1201-202 1 39.70o 217 1 Examples 202 204 Preparation of cycloartenyl, cyclobranyl and 24-methylenecycloartanyl esters of p-nitro-a-methylcinnamic acid Thionyl chloride (112 mt, 4 equivalents) and dimethylformamide (1 mk) were added to p-nitro-amethylcinnamic acid (78.3 g, 0.378 mole) and the mixture was stirred at 60 0 C for 2 hours. Then the resulting mixture was evaporated to dryness under reduced pressure, and the residue was mixed with dioxane (250 mk) and with pyridine (250 m2), and the mixture was allowed to react with cycloartenol (125.0 g, 0.293 mole), cyclobranol (129.1 g, 0.293 mole), or 24-methylenecycloartanol (129.1 g, 0.293 mole) at 60°C for 2 hours. Then, the solvents were distilled off under reduced pressure, and the residue was extracted with chloroform. The extracts were washed successively with water and saturated aqueous solution of sodium bicarbonate, then dried, and evaporated to dryness under reduced pressure. The residual crystals were recrystallized from chloroformethanol giving each of the title ocmpounds.

The yield m.p. and specific rotation 25 {[aD (C 1.00 CHC 1 3 of each product were as follows: D 3 j OG* 00 0 t 09 i 6 f e r. I 2183 1 I I nl 6t~* 4 4) 4I 4* O

C

4 44 4 46 S; I 4t t 4l Example Triterpenyl ester of Yield Melting Specific No. p-nitro- -methyl- point rotation cinnamic acid I25

D

204 Cycloartenyl ester 88.7 188-189 +44.20 205 Cyclobranyl ester 89.5 222-223 +41.70 206 24-Methylene- 88.8 211-212 +43.50 cycloartanyl ester Examples 205 207 Preparation of cycloartenyl cyclobranyl, and 24-methylenecycloartanyl esters of p-amino-a-methylcinnamic acid Each of cycloartenyl, cyclobranyl, and 24methylene-cycloartanyl esters of p-nitro-a-methylcinnamic acid (16.6 g, 17.0 g, and 17.0 g, respectively, 0.027 mole each) obtained in Examples 204 206, respectively, was suspended in a mixture of acetic acid (150 mZ) and dioxane (150 mk), 6N-hydrochloric acid-dioxane (9.5 mZ) and zinc powder (8 g) were added to the suspension and the mixture was stirred at 40 0 C for 3 hours. After the reaction, zinc powder was removed by filtration. The filtrate was evaporated to dryness under reduced pressure. The residue was extracted with chloroform.

The extracts were washed successively with water and saturated aqueous sodium bicarbonate solution, then dried, and evaporated to dryness.. The residual 219 1 crystals were recrystallized from chloroform-ethanol giving each of the title compounds. The yield m.p. and specific rotation (C 1.00, CHC1 3 of each product were as follows: Example Triterpenyl ester of Yield Melting Specific No. p-amino-a-methyl- point rotation cinnamic acid 205 Cycloartenyl ester 91.2 168-169 +42.60 206 Cyclobranyl ester 91.8 202-203 +41.20 24-Methylene- 207 92.0 193-194 +42.30 cycloartanyl ester Ittt t*t t 9 0 48 048 a .9 #0 *r 04 4 I 0 Examples208 210 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of m-nitro-a-methylcinnamic acid Thionyl chloride (60 ma, 2.1 equivalent) and dimethylformamide (1 mk) were added to m-nitro-amethylcinnamic acid (80.4 g, 0.388 mole) and the mixture was stirred at 60 0 C for 2 hours. After concentration of the resulting mixture under reduced pressure, the residue was mixed with dioxane (300 mi) and with pyridine (200 mt), and the mixture was allowed to react with cycloartenol (125.9 g, 0.295 mole), cyclobranol (130.0 g, 0.295 mole), or 24-methylenecycloartanol (130.0 g, 0.295 mole) at 60 0 C for 2 hours. Then the 22 C) 1 mixture was evaporated to dryness under reduced pressure, and the residue was extracted with chloroform. The extracts were washed successively with water and saturated aqueous solution of sodium bicarbonate, then dried, and evaporated to dryness under reduced pressure.

The residual crystals were recrystallized from chloroformethanol v7v), giving each of the title compounds.

The yield m.p. and specific rotation D (C 1.00, CHC1 3 of each product were as follows: Example Triterpenyl ester of Yield Melting Specific No. m-nitro-a-methyl- point rotation cinnamate [a]D 208 Cycloartenyl ester 92.4 161-162 +43.00 209 Cyclobranyl ester 91.7 193-194 +42.30 24-Methylene- 210 90.9 172-173 +42.70 cycloartanyl ester ft t 0 0 t: 4 Examples 211 213 Preparation of cycloartenyl, cyclobranyl, and 24-methylenecycloartanyl esters of m-amino-a-methylcinnamic acid Each of cycloartenyl-, cyclobranyl-, and 24methylenecycloartanyl esters of m-nitro-a-methylcinnamic acid (16.6 g, 17.0 g, and 17.0 g, respectively, 0.027 mole each) obtained in Examples 208 210, respectively, was suspended in a mixture of acetic acid (150 m) and r i

I

2 ""nar~lll-~lr~ 221 1 tetrahydrofuran (200 mi). 6N-hydrochloric acid-dioxane (12.5 mt) and zinc powder (16.5 g) were added to the suspension and the mixture was stirred at 20°C for 2 hours. After the reaction, zinc powder was removed by filtration. The filtrate was concentrated under reduced pressure, and extracted with chloroform. The extracts were washed successively with water and saturated aqueous solution of sodium bicarbonate, then dried, and evaporated to dryness. The residual crystals were recrystallized from chloroform-ethanol giving each of the title compounds. The yield m.p. 25 and specific rotation (C 1.00, CHC1 3 of each product were as follows.

15 Example Triterpenyl ester of Yield Melting Specific No. m-amino- -methyl- point rotation cinnamic acid (oC) [a]D 211 Cycloartenyl ester 85.2 171-172 +44.20 212 Cyclobranyl ester 86.3 198-199 +42.80 24-Methylene- 213 84.2 177-178 +43.40 cycloartanyl ester *4#4 t: S 4I S 0

S..

aiI I 44f 4 441 I 9 t 4414 $4 Examples 214 and 215 Preparation of cycloartenyl and cyclobranyl esters of p-nitro-a-ethylcinnamic acid Thionyl chloride (6 mt, 2.0 equivalent) and i 222 1 dimethylformamide (0.1 mt) were added to p-nitro-aethylcinnamic acid (8.9 g, 0.040 mole) and the mixture was stirred at 60 0 C for 2 hours. Then the resulting mixture was concentrated under reduced pressure. The residue was mixed with dioxane (30 mz) and with pyridine (20 mt), and the mixture was allowed to react with cycloartenol (12.8 g, 0.030 mole) or cyclobranol (13.2 g, 0.030 mole) at 600C for 2 hours.

Then the resulting mixture was concentrated under reduced pressure, and the residue was extracted with chloroform. The extracts were washed successively with water and saturated aqueous solution of sodium bicarbonate, then dried, and evaporated to dryness under reduced pressure. The residual crystals were recrystallized from chloroform-ethanol v/v), giving each of the title compounds. The yield m.p.

and specific rotation 2 5 (C 1.00 CHC 3 of each product were as follows: toot I *11 I t L #4 4C 1( Example Triterpenyl ester of Yield Melting Specific No. p-nitro-a-ethyl- point rotation cinnamic acid (oC) [D 214 Cycloartenyl ester 92.1 178-179 +44.80 215 Cyclobranyl ester 91.8 204-205 +43.70 r' -l, 223 1 Examples 216 and 217 Preparation of cycloartenyl and cyclobranyl esters of p-amino-a-ethylcinnamic acid Each of cycloartenyl and cyclobranyl esters of p-nitro-a-ethylcinnamic acid (17.0 g and 17.4 g, respectively, 0.027 mole each) obtained in Examples 214 and 215, respectively, was suspended in a mixture of acetic acid (150 mZ) and tetrahydrofuran (200 mt).

6N-Hydrochloric acid-dioxane (12.5 mZ) and zinc powder (16.5 g) were added to the suspension and the mixture was stirred at 22 0 C for 2 hours. After the reaction, zinc powder was removed by filtration. The filtrate was evaporated to dryness under reduced pressure, and the residue was extracted with chloroform. The extracts were washed successively with water and saturated aqueous solution of sodium bicarbonate, then t f dried, and evaporated to dryness under reduced pressure.

The residual crystals were recrystallized from chloroformethanol giving each of the title compounds.

The yield m.p. and specific rotation ([3]25 (C 1.00, CHCl 3 of each product were as follows: B i i f2 0 0

III

4 t( 1 224 Example Triterpenyl ester of Yield Melting Specific No. p-amino-a-ethyl- point rotation cinnamic acid 216 Cycloartenyl ester 80.6 160-161 +45.20 217 Cyclobranyl ester 79.3 200-201 +44.30 Example 218 Preparation of 24-methylenecycloartanyl ester of 3-methoxy-4-nitrobenzoic acid The title compound was prepared according to following the procedure of Example 168 where 24-methylenecycloartanol (93.0 g, 0.211 mole was used. The yield was 93.7 g (71.6 m.p. 205 206 0

C.

Specific rotation []25.5 56.5 0 (C 1.00, CHC 3 Analysis, Calcd. for C 39 H57NO 5 (M.W.619.85): C 75.57, H 9.27, N 2.26 Found: C 75.51, H 9.38, N 2.28 Example 219 Preparation of 24-methylenecycloartanyl ester of 4-amino-3-methoxybenzoic acid The title compound was prepared according to the procedure of Example 169 where 24-methylenecycloartanyl ester of 3-methoxy-4-nitrobenzoic acid was used (92.2 g, 0.149 mole) obtained in Example 218. The yield was 62.8 g (71.5 m.p. 222 223 0

C.

Specific rotation []25 63.2 0 (C 1.00, CHCI) D 3 225 1 Analysis, for C39H59NO3(M.W.589.86) Calcd. C 79.41, H 10.08, N 2,37.

Found C 79.38, H 10.14, N 2.35 Example 220 Preparation of cycloartenyl ester of 4amino-3-methoxybenzoic acid 4-Acetamido-3-methoxybenzoic acid (6.5 g, 0.031 mole) in dioxane (110 mz) was allowed to react with thionyl chloride (21.0 ma) and then pyridine (0.5 m9) at 20 0 C, and the mixture was stirred at 50 0 C for minutes. Then the resulting mixture was evaporated to dryness under reduced pressure, and to the residue a solution of cycloartenol (10.0 g, 0.023 mole) in a dioxane (50 mZ)-benzene(50 m9) mixture was added and S" 15 further pyridine (20 mk). After the mixture was S' heated at 70 0 C for 3 hours, the solvents were distilled 44 44 off under reduced pressure. The residue was dissolved *4 in chloroform (100 mZ), and the chloroform solution was washed with saturated aqueous solution of sodium bicarbonate. The aqueous solution was extracted with ,chloroform (5 x 10 The whole chloroform solution was dried and concentrated under reduced pressure, and .o 0 the residue was purified by silica gel column chromatography [solvent: chloroform-ethyl acetate giving cycloartenyl ester of 4-acetamido-3-methoxybenzoic c e acid (10.8 g) in a 76.5 yield. m.p. 224 225 0

C.

25 Specific rotation 61.50 (C 1.00, CHC1 3 D 3 r 226 G 1 Cycloartenyl ester of 4-acetamido-3-methoxybenzoic acid (10.0 g, 0.016 mole) thus obtained was allowed to react with 30 HC% (20 mf) in tetrahydrofuran (200 under reflux for 2 hours. Then the solvent was distilled off under reduced pressure, and the residue was dissolved in chloroform (300 mt), and the chloroform layer was washed successively with IN aqueous NaOH (200 mf) and saturated saline water. The aqueous solutions were extracted with chloroform 3 times. The combined chloroform solution was dried and concentrated and the residue was purified by silica gel column chromatography [solvent: ethyl acetate-hexane giving cycloartenyl ester of 4-amino-3-methoxybenzoic acid (5.4 g) in a 58.7 yield. m.p. 186 187 0

C.

15 26 S' 15 Specific rotation [a]D 64.4 (C 1.00, CHCl 3 t 1 4 4 Example 221 Preparation of cyclobranyl ester of 4amino-3-methoxy-a-methylcinnamic acid 4-Acetamido-3-methoxy-a-methylcinnamic acid S 2 0 (21.93 g, 0.088 mole) dissolved in dioxane (150 mk) I. was allowed to react with thionyl chloride (25.7 m%) t t at 60°C for 2 hours with stirring. Then the solvent l was distilled off under reduced pressure. The residue SIwas dissolved in dioxane (150 mk) and pyridine (50 mP), 25 and allowed to react with cyclobranol (30 g, 0.068 mole) Sat 60°C for 2 hours with stirring. The resulting mixture was concentrated under reduced pressure, and 227 1 ethyl acetate (300 mZ) was added to the residue. The resulting crystals were purified by silica gel column chromatography [solvent: chloroform-ethyl acetate (1:1, giving cyclobranyl ester of 4-acetamido-3methoxy-a-methylcinnamic acid (38.5 g) in a 84.2 yield, m.p. 248 249 0

C.

26+ Specific rotation [a] 6 38.20(C 1.00, CHC13 Cyclobranyl ester of 4-acetamido-3-methoxy-amethylcinnamic acid (34.4 g, 0.051 mole) thus obtained was dissolved in tetrahydrofuran (300 mz), and was allowed to react with 30 HC1 (60 m) at 70 0 C for 2 Lz'irs with stirring. The resulting mixture was concentrated under reduced pressure, and the residue was r purified by silica gel column chromatography [solvent: St 15 chloroform-ethyl acetate, giving cyclobranyl ester of 4 -amino-3-methoxy-a-methylcinnamic acid S (18.9 g) in a 58.8 yield. m.p. 225 226 0

C.

25 Specific rotation [a]25 42.0(C 1.00, CHC1 3 20 Example 222 Preparation of 24-methylenecycloartanyl ester of 4-amino-3-methoxycinnamic acid 2 4-Methylenecycloartanyl ester of 4o propionamido-3-methoxycinnamic acid was prepared according to the procedure of Example 221 where 4propionamido-3-methoxycinnamic acid (21.93 g, 0.088 S' mole) and 24 -methylenecycloartanol (30 g, 0.068 mole) were used in place ofi 4-acetamido-3-methoxy-a-methyl 228 1 cinnamic acid and cyclobranol, respectively. The yield was 38.4 g (83.8 210 211'C.

Specific rotation [a 26 39.40 (C 1.00, CHCl 3 Then 24-methylenecycloartanyl ester of 4-amino- 3-methoxycinnamic acid (18.7 g) was prepared according to the procedure of Example 221 where 24-methylenecycloartanyl ester of 4-propionamido-3-methoxyc innamic acid was used (35.2 g, 0.052 mole) in place of cyclobranyl ester of 4-acetamido-3-methoxy-a-methylcinnanic acid.

The yield was 18.7 g (57.1 227 228'C.

Specific rotation .0 CHCl) D 3 0. Q*

*C

c~,t i~ 229 1 Formula Example 1 Tablets .A Compound of Example 43: Cyclobranyl ester of p-aminobenzoic acid 100 m Mannitol 123 Hydroxypropoxymethylcellulose' 7 Talc 5 Microcrystalline cellulose Hydrogenated castor oil 5 g I r c, t: r(rr Tablets B Compound of Example 86: 24-Methylenecycloartanyl ester of 4-hydroxy-3-methoxybenzoic acid Corn starch Lactose Talc Magnesium stearate Tablets C Compound of Example 100-2: Cyclobranyl ester of 4-hydroxy-3methoxycinnamic acid Soluble starch Corn starch Microcrystalline cellulose Silicon dioxide Magnesium stearate Total 300 mg 150 mg 160 180 7 3 Total 500 mg r~~ fjr jc cc ttr cr 4- 4, r4 4 44 4 44 ccr c 100 mg 125 45 6 4 i Total 300 mg 230 1 Tablets D Compound of Example Cycloartenyl ester of 3-ethoxy-4- 100 mg hydroxybenzoic acid Lactose 147 Corn starch 62.1 SMicrocrystalline cellulose 90 Magnesium stearate 0.9 Total 400 mg Tablets E Compound of Example 71: Cyclobranyl ester of 3-ethoxy-4- 50 mg hydroxycinnamic acid Lactose 10 Microcrystalline cellulose 85.5 15 Carboxymethyl cellulose calcium 2 Magnesium stearate 1.5 Stearic acid 1 Total 150 mg According to the above prescriptions, various weights tablets were made of thoroughly uniformly mixed Spowders with a tableting machine.

SIn addition, tablets were prepared according Sto the recipe above but substituting, cycloartenyl ester of 4-hydroxy-3-methoxy-a -methylcinnamic acid Sof Example 114 for cyclobranyl ester of p-aminobenzoic n 25 Etr acid of Example 43; 24-methylenecycloartanyl ester of 4-hydroxy-3-methoxy- -methylcinnamic acid of Example 4-hydroxy-3-methoxy- a-methylcinnamic acid of Example 4i I: i i i 231.

1 118 for 24-methylenecycloartanyl ester of 4-hydroxy- 3-methoxybenzoic acid of Example 86; cyclobranyl ester of 4-hydroxy-3-methoxy- a-methylcinnamic acid of Example 116 for cyclobranyl ester of 4-hydroxy- 3-methoxycinnamic acid of Example 100-2; cycloartenyl ester of p-amino- a-methylcinnamic acid of Example 205 for cycloartenyl ester of 3-ethoxy-4-hydroxybenzoic acid of Example 60; and cyclobranyl ester of 2-methoxybenzoic acid of Example 173 for cyclobranyl ester of 3-ethoxy-4-hydroxycinnamic acid of Example 71.

Formula Example 2 Granules A Compound of Example 19: Cycloartenyl ester of p-aminobenzoic acid 100 mg Lactose 22 Microcrystalline cellulose 60 Corn starch 15 Hydroxypropylcellul,6se 3 Total 200 According to the above prescription, cycloartenyl ester of p-aminobenzoic acid, lactose, microcrystalline cellulose, and corn starch were mixed together. The mixture was sprayed with a 5 aqueous solution of hydroxypropylcellulose as a binder, dried and granulated by using a fluidized bed granulator.

a..

a Iaaa

I

**a *r S a a t t at S at 4 4 4 9

S*

rr ;Ijn"~

I

I*

a 4444 44 a.

*i 4.I I U 232 1 Another granules were prepared in the same method as the above recipe but substituting cycloartenyl ester of 4-amino-3-methoxybenzoic acid of Example 167 for cycloartenyl ester of p-aminobenzoic acid of.Example 19.

Formula Example 3 Granules B Compound of Example 43: Cyclobranyl ester of p-aminobenzoic acid 100 mg 1 Hydroxypropy1cellulose 2 1 0 Total 200 mg r According to the above prescription, the Mannitol 38 cyclobranyl ester mannitol, microcrystalline cellulose 48 Potato starch 10 potato starch and polyvinylpyrrolidone were mixed together, then the mixture was sprayed with 5 0 aqueous solution ydroxypropylcellulose as a binder, dried and granulated, by using a fluidized 0 0 ootbed granulator.

Another granules were prepared in the same meth od as the above recipto the above prescription, thcycloartenyl ester of 5-amino-2-propoxy-a -methylcinnamic acid of Example 195 for cyclobranyl estalline cellulose, potato starch and polyvinylpyrrolidone were mixed together, then the mixture was sprayed with 5 aqueous solution of hydroxypropylcellulose as a «binder, dried and granulated, by using a fluidized bed granulator.

r om •Another granules were prepared in the same method as the above recipe but substituting cycloartenyl ester of 5-amino-2-propoxy-a -methylcinnamic S9* acid of Example 195 for cyclobranyl ester of

%.L

i, e 233 1 p-aminobenzoic acid of Example 43.

Formula Example 4 Granules C Cyclobranol Mannitol Microcrystalline cellulose Carboxymethyl cellulose calcium Magnesium stearate In Hardened oil 100 mg 2 Total 200 mg 4r o o 4 4*4 1'44 4 *4 4on 0 4 fl 4 Granules D Compound of Example 24: Cycloartenyl ester of nicotinic acid Corn starch Microcrystalline cellulose Carboxymethyl cellulose calcium 100 mg 29 21 Total 200 mg 2 p i i 40 *o 4 444 4 *0 *i 4 .4 4 4 O p .4.

Granules E Compound of Example 49: Cyclobranyl ester of m-aminobenzoic acid 100 mg Lactose 53 Corn starch 39 Potato starch 2 Talc 3 Magnesium stearate 3 Total 200 mg r ;:ii~b i; i I l t- 'l-r~ll l 234 1 According to the above prescriptions, the ingredients were mixed uniformly and granulated with an extruder.

In addition, another granules were prepared in the same method as the above recipe but substituting; cycloartenyl ester of 4-hydroxy- a-ethylcinnamic acid of Example 130 for cyclobranol; cycloartenyl ester of 4-amino-3-methoxycinnamic acid of Example 177 for cycloartenyl ester of nicotinic acid of Example 24 and cycloartenyl ester of 5-amino-2-methoxybenzoic acid of Example 171 for cyclobranyl ester of m-aminobenzoic acid of Example 49.

4 4 I I 1414 Formula Example Capsules A Compound of Example 100-2: Cyclobranyl ester of 4-hydroxy-3-methoxy- 100 cinnamic acid Lactose 28 Microcrystalline cellulose 47 Mannitol Corn starch Polyvinylpyrrolidone 2 Hydroxypropylcellulose 3 mg it It It to 4r I 4 *1 iC *4: *4 q 4 4o4 S *5 Vt *C 4 S a Total 200 mg According to the above prescription, cyclobranyl ester of 4-hydroxy-3-methoxycinnamic acid, lactose, microcrystalline cellulose, mannitol, corn 235 1 starch and polyvinylpyrrolidone were mixed together, then the mixture was sprayed with a 5 aqueous solution of hydroxypropylcellulose as a binder, dried and granulated, by using a fluidized bed granulator.

No. 3 hard capsules were each filled with 200 mg of these granules.

Another hard capsules were prepared in the same method as the above recipe but substituting cycloartenyl ester of 4-hydroxy-3-methoxy- amethylcinnamic acid of Example 114 for cyclobranyl ester of 4-hydroxy-3-methoxycinnamic acid of Example 100-2.

Formula Example 6 Capsules B Hard capsules were prepared by filling each of No. 3 hard capsules with 160 mg of granules A K, prepared according to Formula Example 2.

Formula Example 7 Capsules C Hard capsules were prepared by the following procedure. No. 2 hard capsules were filled with 200 mg of granules D prepared in Formula Example 4.

St 23 1Formula Example 8 Capsules D Compound of Example 12: Cycloartenyl ester of m-hydroxybenzoic acid 100 mg Mannitol 98 Carboxymethyl cellulose calcium 2 Total 200 mg According to the above prescription, the ingredients were mixed uniformly. Gelatin capsules of No. 2 were filled with 200 mg of the mixed powder each. Then the capsules were enteric-coated to give enteric capsules.

Another enteric capsules were prepared by the same method as the above recipe but substituting cycloartenyl ester of 4-hydroxy-3-propoxy-a methylcinnamic acid of Example 146 for cycloartenyl ester of m-hydroxybenzoic acid of Example 12.

Formula Example 9 Capsules

E

Compound of Example 58: Cyclobranyl ester of p-hydroxybenzoic acid 200 g Sodium laurylsulfate 9 Disodium hydrogen ,phosphate 1 Mannitol 188 Magnesium stearate 2 Total 400 g I *r a a,.

cv *r 9 4, ,I ar ar 4 237 1 According to the above prescription, ingredients were uniformly mixed. Gelatin capsules of No. 1 were filled with 300 mg of the mixture each. Another capsules were prepared by the same method as the above recipe but substituting cyclobranyl ester of m-amino-a -methylcinnamic acid of Example 212 for cyclobranyl ester of p-hydroxybenzoic acid.

Formula Example Enteric granules Compound of Example Cycloartenyl ester of linoleic acid 100 g Mannitol 16 Microcrystalline cellulose 65 Corn starch 15 Hydroxypropylmethylcellulose 3 Vinylpyrrolidone-vinyl acetate 1 copolymer (supplied by General Aniline Film Corp.) Total 200 g According to the above prescription, the ingredients were uniformly mixed and then granulated into spherical granules by an extruder. These granules were coated with material composed of hydroxypropylmethylcellulose phthalate (74 glyceryl triacetate (11.6 stearic acid (11.6 and light silicon dioxide (2.8 to give enteric granules.

r a:i i :i I 238 Another enteric granules were prepared in the same method as the above recipe but substituting cycloartenyl ester of 5-amino-2-methoxybenzoic acid of Example 171 for cycloartenyl ester of linoleic acid of Example Formula Example 11 Enteric tablets Compound of Example 54: Cyclobranyl ester of m-hydroxybenzoic acid 100 mg Mannitol 10 Microcrystalline cellulose Carboxymethyl cellulose calcium 2 Magnesium stearate 1.5 Hardened oil 1.5 Total 200 mg According to the above prescription, the ingredients were mixed uniformly. The mixture was compressed into tablets with a tabletting machine, followed by coating with the substance for enteric coating below.

Coating substance: Hydroxypropylmethylcellulose phthalate 14.8 mg Dioctyl phthalate 2.3 25 Stearic acid 2.3 Light silicon dioxide 0.6 Total 20 mg

C

I Ji I4 L S.t CI 4 t44 4S4 i .4'.l 4* *4 a. i

S

I i i I i- 233 1 In addition, enteric tablets were prepared in the same method as the above recipe but substituting cycloartenyl ester of 5-amino-2-ethoxycinnamic acid of Example 183 for cyclobranyl ester of m-hydroxybenzoic acid of Example 54.

Formula Example 12 Granules Compound of Example 16: Cycloartenyl ester of o-nitrobenzoic acid 100 mg Corn starch 32.5 Hydroxypropylcellulose Total 136 mg According to the above prescription, the 15 ingredients were uniformly mixed, the mixture was nucleated q oe S° by tumbling or centrifugal methods. Then these nuclei S were diluted with the uniformly mixed excipient below.

A

.r The diluted nuclei were allowed to adhere each other with a usual binder; then coated and granulated. The granules (230 mg) were coated with the same enteric coating substance (30 mg) as shown in Formula Example t 11. No. 3 gelatin capsules were each filled with 260 mg of the enteric granules.

t Excipient: 5 Granulated sugar 33.0 mg Corn starch 58.5 t I Hydroxypropylcellulose 2.5 Total 94 mg I 240 1 Formula Example 13 Powders A Compound of Example 100-1: Cycloartenyl ester of 4-hydroxy-3methoxycinnamic acid Mannitol Corn starch 100 mg Total 200 mg Powders B Compound of Example 46: Cyclopranyl ester of o-aminobenzoic acid 100 mg Corn starch 100 Total 200 mg Powders were prepared by mixing the above 15 ingredients uniformly in a double cone type blender.

I4 fa a a I a a b a, f \t c..

Formula Example 14 Capsules Compound of Example 93: 24-Methylenecycloartanyl ester of linoleic acid Vitamin C Citric acid Carboxymethyl cellulose calcium Sodium laurylsulfate Polyoxyethylene monostearate Methylene chloride 260 g 10 5 5 300 ml tro 4r 4 r

I

i i r 1 The above ingredients were thoroughly mixed to make a suspension. Aerosil 200 400 (tradename) (180 g) was added to the suspension, then the mixture was stirred and dried. The resulting solid mass was ground to a powder, and a chlorothene-ethanol solution (300 ml) which contains 20 g of vinylpyrrolidone and 2 acid methyl acrylate copolymer as a binder was added to the powder. The mituare was kneaded and granulated with a pelleter according to the ordinary method, and the granules were dried at about 50 °C (containing about 51 24-methylenecycloartanyl ester of linoleic acid). Hard capsules were filled with 200 mg of the granules with mixing a small amount of magnesium stearate each by using an automatic capsule-filling machine.

itt if I ,r c c t ft t

I

LI (C LI I Formula Example Suppositorys A Compound of Example 74: Cycloartenyl ester of 4-hydroxy-3propoxycinnamic acid Witepsol E-85 W'itepsol W-35 Methyl-p-hydroxybenzoate Butyl-p-hydroxybenzoate a 200 mg 540 1454 3 3 Total 2200 mg i 2A 4 2 Suppositorys B Compound of Example 29: Cycloartenyl ester of 4-hydroxy-3methoxybenzoic acid Ascorbic acid Polyethylene glycol 200 mg 2080 Total 23flW o p 0000 ~p o 000400 0 #0 00 00 0 0 *000 #0*4 *0 p I p *0

I

00 0 10 I 4 0 top 0040 0 0 ro 00 r' c r e Suppositorys C Compound of Example 39: Cyclobranyl ester of 3,4-dihydroxybenzoic 200 mg acid Butylhydroxyanisole 6 Semisythetic glyceride 2900 Total 3106 mg Suppositorys D Compound of Example 27: Cycloartenyl ester of 3,4-dihydroxybenzoic 200.0 mg acidl

A

7fl Gelatin Glycerol Methyl- p -hydroxybenzoate Propyl -p-hydroxybenzoate Ethylvanillin Titanium dioxide DC yellow lake No. 5 Distilled water 1152.0" 454 .0" 0.4 2.8 24 .0 12.0 352.8" Total 2200.0 mg 2- 43 J 243 1 According to the above prescriptions, the ingredients were thoroughly mixed and melted.

And the melts were cast in aluminum molds, and cooled to give suppository.

Formula Example 16 Emulsion Polysolvate 80 (tradename, 1 g) and DK Ester F- 160 (tradename, 1 g) were dissolved in a 50 aqueous solution of (100 .ml) sorbitol. Cyclobranyl ester of 4-acetoxy-3-methoxycinnamic acid (50 g) was added thereto and the mixture was emulsified by stirring 04 o .po at 80 OC for 30 minutes. Sodium benzoate (0.25 g) and citric acid (1 g) were dissolved therein, then the volume was adjusted to 500 ml with distilled 1 t, water. The whole was stirred again, giving the emulsion (containing about 10 cyclobranyl ester of 4-acetoxy-3-methoxycinnamic acid).

t I It 1 t Formula Example 17 Tablets F Compound of Example 102: 'Cycloartenyl ester of p-aminocinnamic 00 mg Stacid SMannitol 123 Hydroxypropoxymethylcellulose 7 Talc 5 i) i II R i I- a I a -r Isr L1- -n L- 244 Microcrystalline cellulose Magnesium stearate 61 mg 4 Total 300 mg Tablets G Compound of Example 104: Cyclobranyl ester of p-aminocinnamic ac-id Lactose Corn starch 100 mg 147 62 Microcrystalline cellulose Magnesium stearate To; *4*r #444 4. 4* *o U 4c 4 4 44 4 1 4 84 EP I 4r 44- C 4 Tablets H Compound of Example 108: Cycloartenyl ester of m-aminocinnamic acid Soluble starch Corn starch Microcrystalline cellulose Silicon dioxide 86 5 tal 400 mg 100 mg 125 45 6 i Magnesium stearate 4 Total 300 mg According to the each prescription above, the ingredients were uniformly mixed and compressed into the tablets of each weight by using a tabletting machine.

-I

245 1 Formula Example 18 Granules F Compound of Example 112: 2 4-Methylenecycloartanyl ester of m-aminocinnamic acid 100 mg Lactose 22 Microcrystalline cellulose Corn starch Hydroxypropylcellulose 3 Total 200 mg According to the prescription, 24-methylenecycloartanyl ester of m-aminocinnamic acid, lactose, microcrystalline cellulose, and corn starch were mixed. The mixture sprayed with a 5 aqueous hydroxypropylcellulose solution as a binder was dried and granulated with a fluidized bed granulator.

c p ra o it'

I.

IIt Ct I 1 *l 41 Ii

I

Formula Example 19 Granules G Compound of Example 102: Cycloartenyl ester of p-aminocinnamic acid Lactose Corn starch Potato starch Talc Magnesium stearate To1 100 mg 53 39 2 3 3 tal 200 mg pop-- I IBIIIU 246 According to the above prescription, the ingredients were uniformly mixed and granulated with an extruder.

Formula Example Capsules F Hard capsules of No. 2 were filled with granules G prepared according to Formula Example 19.

One capsule contained 200 mg of granules G.

O**

a a

II

0 a 0 eoC *a i ia

OC

*0 Formula Example 21 Capsules G Compound of Example 102: Cycloartenyl ester of p-aminocinnamic acid Sodium laurylsulfate Disodium hydrogen phosphate Mannitol Magnesium stearate

T

150 mg 4 1 93 2 otal 250 mg According to the above prescription, the ingredients were uniformly mixed. No. 1 gelatin capsules were filled with 250 mg of the mixed powder each.

0 sa..

CC 90 CI S

C

Claims (10)

1. A triterpenyl ester derived from triterpenyl alcohol and organic acid other than ferulic acid and monobasic and dibasic saturated fatty acids, wherein the triterpenyl alcohol is selected from the group consisting of cycloartenol, cyclobranol, 24-methylenecycloartanol, lanosterol, lanostenol, agnosterol, cyclosadol, dihydroagnosterol, cyclolaudenol, cycloartanol, cycloeucalenol, euphol, butyrospermol, tirucallol, euphorbol and dammerdienol and the organic acid is nicotinic acid, linoleic acid, or an organic acid of the formula Ar(CH=CR) -COOH wherein R is hydrogen or a C 1 -C 4 -alkyl group, and Ar is aminophenyl, nitrophenyl, hydroxyphenyl, a C 1 -C 4 -alkoxyphenyl, C 1 -C 4 -alkyl-CONHphenyl, (C 1 -C 5 -alkyl)C00-phenyl, or Ar is a phenyl substituted by C 1 -C 4 -alkoxy and I hydroxy, by C 1 -C 5 -alkyl-C and hydroxy, by C 1 -C 4 alkoxy and (C 1 -C 5 alkyl)C00, by C 1 -C 4 -alkoxy and nitro, by C 1 -C 4 -alkoxy and amino, or by C 1 -C 4 -alkyl-CONH and C 1 -C 4 -alkoxy or Ar is di-(C 1 -C 4 -akoxy)pheny, di(C 1 -C 5 -alkyl-C0)phenyl or dihydroxyphenyl; and n is 0 or 1.

2. The ester of organic acid according to Claim 1, wherein the organic acid is a substituted cinnamic acid, benzoic acid, or a-(C 1 -C 4 alkyl) cinnamic acid substituted by one member selected from the group consisting of amino, nitro, hydroxyl, C1-C4 alkoxy, C2-Cg acylamino, and C2-C6 alkylcarboxyl groups on the benzene ring.

3. The ester of organic acid according to Claim 1, wherein the organic acid is a substituted cinnamic acid, benzoic acid, or cx-(C 1 -C 4 alkyl) cinnamic acid having two substituents on the benzene ring, said substituents being one pair selected from the group consisting of hydroxyl and C 1 -C alkoxy groups, or1 4 cl*0' hydroxyl and C2-C6 alkylcarboxyl groups, C1-C4 alkoxy and Cz-C6 it'; alkoxycarboxyl groups, C -C4 alkoxy and nitro groups, C 1 -C 4 alkoxy and amino groups, I It 4 1 TMR/866c 243 1- 4 alkoxy and C 2 C 5 acylamino groups, two C1- C 4 alkoxy groups, two C 2 -C 6 alkyl carboxyl groups, and two hydroxyl groups. 4 The ester of organic acid according to Claim 1, wherein the organic acid is nicotinic acid or linoleic acid. The ester of organic acid according to Claim, which is a cycloartenyl ester or cyclobranyl ester of p- acetoxycinnamic acid, p-hydroxycinnanic acid, in-, or p-nitro- cinnamic acid, in-, or p-arninocinnamic acid, in-, or p- acetoxybenzoic acid, mn-, or p-hydroxybenzoic acid, m or p-rethoxybenzoic aim,- rpntoezi acid, in-, or p-aminobenzoic acid, p-acetamidobenzoic o -mthycinnmicacid, 3- or 4-propionyloxy-o<-nethylcinnanic 3- or p-nitrox-O(-nethylcinnamic acid, or p-io ethylcinnamic acid, 2-hydroxy-Q(-methylcinnanic acid, p-nitro-c-ethylcinnanic aci&:, or p-ainino-c(-ethylcinnaiic acid. t It 4 The ester of organic acid according to Claim A' which is a cycloartenyl ester or cycJlobranyl ester of 3 ,4-diacetoxycinnarnic acid, 3 ,4-dihydroxycinnamic acid, 3 ,4-dipropionyloxycinnamic acid, 3 ,4-diiethoxycinnamic acid, 4-acetoxy-3-inethyoxycinnamic acid, 4-acetoxy-3- ethoxycinnamic acid, 3-ethoxy-4-hydroxycinnamic acid,

24.9 1 4-hydroxy-3-n-propoxycinnamic acid, 3-methoxy-4-nitro- cinnamic acid, 4-arnino-3-methoxycinnamic acid, 2-ethoxy- acid, 5-amino-2-ethoxycinnamic acid, 3 ,4-diacetoxybenzoic acid, 3 ,4-dihydroxybenzoic acid, 3 ,4-dimethoxybenzoic acid, 4-acetoxy-3-methoxybenzoic acid, 4-hydroxy-3--methoxybenzoic acid, 4-acetoxy-3- ethoxybenzoic acid, 3-ethoxy-4-hydroxybenzoic acid, 3- methoxy-4-nitrobenzoic acid, 4-amino-3-methoxybenzoic acid, acid, 5-amino-2-methoxybenzoic acid, 3-methoxy--4-propionyloxy-(X-methylcinnamic acid, 4-hydroxy-3-methoxy-(-methylcinnamic acid, 4-butyryloxy- 3-methoxy-(-ethylcinnamic acid, 4-hydroxy-3-methoxy- X- ethylcinnamic acid, 4-hydroxy-3-methoxy-c/--butylcinnamic acid, 3-ethoxy-4-hydroxy-Q(-methylcinnamic acid, 3-ethoxy- ,-hydroxy-(X-propylcinnamic acid, 3-methoxy-4-nitro-o%- methylcinnamic acid, 4-amino-3-methoxy-(.-methylcinnamic acid, 5-nitro-2-propoxy-.1-methylcinnamic acid, 2-propoxy-Q(-methylcinnamic acid, 3-methoxy-4-nitro-o(- isopropylcinnanic acid, or 4-amino-3-methoxy-oX-isopropyl- a, cinnamic acid. The ester of organic acid according to Claim total, which is the 24-methylenecycloartanyl ester of p-acetoxycinnamic acid, p-hydroxycinnamic acid, m- or p-nitrocinnamic acid, m- or p-aminocinnamic acid, mn-, o- or p-hydroxybenzoic acid, in-, or p-nitrobenzoic acid, or p-aminobenzoic acid, m-acetoxybenzoic acid, 2950 1 m- or p-nitro-o(-methylcinnamic acid, m- or p-amino-Oa- methylcinnamic acid, 3- or 4-propionyloxy-D-methylcinnamic acid, 3- or 4-hydroxy-a-methylcinnamic acid, 3-butyryloxy- 0(-ethylcinnamic acid, or 3-hydroxy-O(-ethylcinnamic acid. The ester of organic acid according to Claim Al, which is the 24-methylenecycloartanyl ester of 4-acetoxy- 3-methoxycinnamic acid, 4-acetoxy-3-ethyoxycinnamic acid, 3-ethoxy-4-hydroxycinnamic acid, 3 ,4-diacetoxycinnamic acid, 3,4-dihydroxycinnamic acid, 3-methoxy-4-nitro- cinnamic acid, 4-amino-3-methoxycinnamic acid, 2-ethoxy- 5-nitrocinnamic acid, 5-amino-2-ethoxycinnamic acid, 4-acetoxy-3-methoxybenzoic acid, 4-hydroxy-3-methoxybenzoic acid, 4-acetoxy-3-ethoxybenzoic ccd, 3-ethoxy-4-hydroxy- benzoic, acid, 3,4-diacetoxyrbenzoic- acid, 3,4-dihydroxy- benzoic acid, 3-methcxy-4-n,,tr.obenzoic acid, 4-amino-3- methoxybenzoic acid, 3-meothoxy-4-propionyloxy-k-methyl- cinnamic acid, 4-hydroxy-3-methoxy--methylc.innanic acid, o 4-butyryloxy-3-methoz, .'X-thylcinnamic acid, 4-hydroxy- 3-methoxy-O -ethy.lcinnamic acid, 4-hydroxy-3-methoxy- butylcinnamic acid, 3-ethoxy-4-hydroxy-o(-methylcinnamic acid, 4-hydroxy-3-propoxy-Q(-propylcinnamic acid, 3-methoxy- 4-nitro--o(-methylcinnamic acid, 4-amino-3-methoxy-- methylcinnamic acid, 5-nitro-2-propoxy-CK-methylcinnamic acid, or 5-amino-2-propoxy-o -methylcinnamic acid. -251- 9, The ester of organic acid according to Claim 3, which is the cycloartenyl ester of 3-methoxy-4-propionyloxycinnamic acid, 3-methoxy-4- i valeryloxy-ax-propylcinnamic acid, 4-hydroxy-3--methoxy-cx-propylcinnamic acid, 4-capryloxy-3-methoxy--butylcinnamic acid, 3ehx--yrx-c ethyl cinnarni c acid, 3-ethoxy-4-hydroxy-cx-butylcinnamic acid, 4-hydroxy- 3-propoxy-ax-methylcinnaiic acid, or 4-hydroxy-3-butoxy-cX-me thylIc innami c acid. The ester of organic acid according to Claim 3, which is the cyciobranyl ester of 4-hydroxy-3-propoxy-a-ethylcinnamic acid. 11. A process for producing a triterpenyl ester derived from triterpenyl alcohol and organic acid other than ferulic acid and monobasic and dibasic saturated fatty acids, wherein the triterpenyl alcohol is selected from the group consisting of cycloartenol, cyclobranol 24-methylenecycloartanol lanosterol, j lanostenol, agnosterol, cyclosadol, dihydroagnosterol, cyclolaudenol, cycloartanol, cycloeucalenol, euphol, butyrospermol, tirucallol, euphorbol and dammerdienol and the organic acid is nicotinic acid, linoleic acid, or an organic acid of the formula Ar(CH-CR) nCO wherein R is hydrogen or a C 1 -C 4 alkyl group, and Ar is aminophenyl, nitrophenyl hydroxyphenyl a C 1 -C 4 alkoxyphenyl C 1 -C 4 alkyl-CONHphenyl, 1- 4-1- 4 (C -C alkyl)COO-phenyl, or Ar is a phenyl substituted by C C -alkoxy and sets hydroxy, by C 1 -C 5 alkyl-COO and hydroxy, by C-C 4 alkoxy and (C alkyl)COO, by C 1 -C 4 alkoxy and nitro, by C 1 -C 4 alkoxy and amino, or by t: I "I C 1 -C 4 alkyl-CONH and C 1 -C 4 alkoxy or Ar is di-(C 1 -C 4 alkoxy)phenyl, dl(C 1 -C -alkyl-COO)phonyl or dihydroxyphenyl; and n is 0 or 1. TMR/866c 9u~; iiil-i~iLi~i~ i i ICI i _liC-i __I 252 which comprises the reaction of the triterpenyl alcohol with an acid halide of the corresponuing organic acid. 12. The process according to Claim 11, wherein the acid halide is that of a substituted cinnamic acid, benzoic acid, or a-(C 1 -C 4 alkyl) cinnamic acid substituted by one member selected from the group of nitro, C 2 -C 5 acylamino, C -C 4 alkoxy, and C 2 -C 6 alkylcarboxyl group on the benzene ring. 13. The process according to Claim 11, wherein the acid halide is that of a substituted cinnamic acid, benzoic acid or a-(C -C 4 alkyl) cinnamic acid having two substituents on the benzene ring, said substituents being one pair selected from the group consisting of C -C 4 alkoxy and C 2 -C 6 alkylcarboxyl groups, C 1 -C 4 alkoxy and nitro groups, C 1 -C 4 alkoxy and C 2 -C 5 acylamino groups, two CI-C 4 alkoxy groups, and two C 2 -C 6 alkylcarboxyl groups. 14. The process according to Claim 11, wherein the acid halide is that of nicotinic acid or of 11noleic acid. A process for producing a triterpenyl ester derived from triterpenyl alcohol and a substituted cinnamic acid, benzoic acid or a-(C 1 -C 4 alkyl) cinnamic acid having a hydroxyl or an amino group, a C 1 -C 4 alkoxy group and a hydroxyl group, a C 1 -C 4 alkoxy group and an amino group, or two hydroxyl groups on th, benzene ring; wherein the triterpenyl alcohol is selected from the group consisting of cycloartenol, cyclobranol, 24--methylenecycloartanol, lanosterol, lanostenol, agnosterol, cyclosadol, dihydroagnosterol, cyclolaudenol, cycloartanol, cycloeucalenol, euphol, butyrospermol, tirucallol, euphorbol and dammerdienol; ,1 Ii i ,i tft t I1 Id I t4 TMR/866c i i 253 which comprises deacylating the triterpenyl ester of the corresponding substituted cinnamic acid, benzoic acid, or a-(C 1 -C 4 alkyl) cinnamic acid having a C 2 -C 5 acylamino or a C 2 -C 6 alkylcarboxyl group, a C 1 -C 4 alkoxy group and a C2-C6 alkylcarboxyl group, a C -C 4 alkoxy group and a C 2 -C 5 acylamino group, or two C2-C6 alkylcarboxyl groups on the benzene ring. 16. A process for producing a triterpenyl ester derived from triterpenyl alcohol and a substituted cinnamic acid, benzoic acid, or a-(C -C 4 alkyl) cinnamic acid having an amino group or an amino group and a C -C4 alkoxy group on the benzene ring; wherein the triterpenyl alcohol is selected from the group consisting of cycloartenol, cyclobranol, 24-methylenecycloartanol, lanosterol, lanostenol, agnosterol, cyclosadol, dlhydroagnosterol, cyclolaudenol, cycloartanol, cycloeucalenol, euphol, butyrospermol, tirucallol, euphorbol and dammerdienol; which comprises 1rt reducing the corresponding ester having a nitro group in place of the amino group, with a metal-acid system. 17. A pharmaceutical composition for the treatment of hyperlipidemia comprising a pharmaceutical carrier and an effective amount of a triterpenyl ester derived from triterpenyl alcohol and organic acid other than ferulic acid and monobasic and dibasic saturated fatty acids, wherein the triterpenyl alcohol is selected from the group consisting of I cycloartenol, cyclobranol, 24-methylenecycloartanol, lanosterol, lanostenol, agnosterol, cyclosadol, dihydroagnosterol, cyclolaudenol, Sc r cycloartanol, cycloeucalenol, euphol, butyrospermol, tirucallol, euphorbol and dammerdlenol and the organic acid is nicotinic acid, linoleic acid, or an organic acid of the formula TMR/866c 254 Ar(CH-CR) -COOH wherein R is hydrogen or a C 1 -C 4 -alkyl group, and Ar is aminophenyl, nitrophenyl, hydroxyphenyl, a C 1 -C 4 -alkoxyphenyl, C 1 -C 4 -alkyl-CONHphenyl, (C -C 5 -alkyl)COO-phenyl, or Ar is a phenyl substituted by C 1 -C 4 -alkoxy and hydroxy, by C 1 -C 5 -alkyl-COO and hydroxy, by C 1 -C 4 alkoxy and (C 1 -C 5 alkyl)CO0, by C 1 -C 4 -alkoxy and nitro, by C -C 4 -alkoxy and amino, or by C 1 -C 4 -alkyl-CONH and C 1 -C 4 -alkoxy or Ar is di-(C -C 4 -alkoxy)phenyl, di(C 1 -C 5 -alkyl-COO)phenyl or dihydroxyphenyl; and n is 0 or 1. 18. The pharmaceutical composition according to Claim 17, wherein the organic acid is a substituted cinnamic acid, benzoic acid, or a-(C,-C 4 alkyl) cinnamic acid substituted by one member selected from the group consisting of amino, nitro, hydroxyl, C -C 4 alkoxy, C 2 -C 5 acylamino, and C 2 -C 6 alkylcarboxyl groups on the benzene ring. 19. The pharmaceutical composition according to Claim 17, wherein the organic acid is a substituted cinnamic acid, benzoic acid, or a-(C 1 -C 4 alkyl) cinnamic acid having two substituents on the benzene ring, said substituents being one pair selected from the group consisting of hydroxyl and C 1 -C 4 alkoxy groups, hydroxyl and C 2 -C 6 alkylcarboxyl groups, C 1 -C 4 alkoxy and C2-C 6 alkylcarboxyl groups, C 1 -C 4 alkoxy and nitro groups, C 1 -C 4 alkoxy and amino groups, C 1 -C 4 alkoxy and C 2 -Cg acylamino group;, two C 1 -C 4 alkoxy groups, two C 2 -C 6 alkylcarboxyl groups, and two hydroxyl groups. The pharmaceutical composition according to Claim 17, wherein the organic acid is nicotinic acid or linoleic acid. t t TMR/866c ,pecC -255- 21. The pharmaceutical composition according to Claim 18, wherein the active ingredient is a cycloartenyl ester or a cyclobranyl ester of p-acetoxycinnamic acid, p-li,ydroxycinnamic acid, m- or p-nitrocinnamic acid, m- or p-aminocinnamic acid, mn-, or p-acetoxybenzoic acid, mn-, or p-hydroxybenzoic acid, in-, or p-inethoxybenzoic acid, in-, or p-nitrobenzoic acid, in-, or p-aininobenzoic acid, p-acetainidobenzoic acid, in- or p-nitro-oc-iethylcinnamic acid, in- or p-amino-X-inethyl- cinnainic acid, 3- or 4-propionyloxy-L-inethylcinnamic acid, 3- or 4-hydroxy-a-methylcinnamic acid, 3- or 4-butyryloxy-c-ethylcinnamic acid, 3- or 4-hydroxy-a-ethylcinnanic, 2-hydroxy-ca-methylcinnanic acid, p-nitro-cc-ethylcinnanic acid, or p-ainino-c-ethylcinnamic acid. 22. The pharmaceutical composition according to Claim 19, wherein the active ingredient is a cycloartenyl ester or a cyclobranyl ester of 3,4-diacetoxycinnanic acid, 3,4-dihydroxycinnanic acid, 3,4-dipropionyloxy- cinnainic acid, 3,4-dimethoxycinnanic acid, 4-acetoxy-3-ethoxycinnanic acid, 3-ethoxy-4-hydroxycinnamic acid, 4-hydroxy-3-propoxycinnamic acid, f3-methoxy-4-nitroclnnamic acid, 4-aiino-3-methoxycinnamic acid, nitrocinnanic acid, 5-amino-2-ethoxycinnanic acid, 3,4-diacetoxybenzolc acid, 3,4-dihydroxybenzoic acid, 3,4-diinethoxybenzoic acid, j 4-acetoxy-3-inethoxybenzoic acid, 4-hydroxy-3-inethoxybenzoi c t CL TMR/866c 256 1 acid, 4 -acetoxy-3-ethoxybenzoic acid, 3-ethoxy-4-hydroxy- benzoic acid, 3-methoxy--4-nitrobenzoic acid, 4-amino-3- methoxybenzoic acid, 2-methoxy-5-nitrobenzoic acid, 2-methoxybenzoic acid, 3-methoxy-4-propionyloxy-O\'-methyl- cinnamic acid, 4 -hydroxy-3-methoxy-o(-methylcinnamic acid, 4 -butyryloxy-3--methoxy-D-ethylcinnamic acid, 4-hydroxy- 3 -methoxy-(:-ethylcinnamic acid, 4-hydroxy-3-methoxy-\- butylcinnamic acid, 3 -ethoxy-4-hydroxy-o -methylcinnamic acid, 3 -ethoxy-4-hydroxy-o -propylcinnami'c acid., 3-methoxy- 4 -nitro-O(-methylcinnamic acid, 4-amino-3-methoxy-O- methylcinnamic acid, 5-nitro-2-propoxy-c(-methylcinnamic acid, 5-amino-2-propoxy-V-methylcinnamic acid, 3-methoxy- 4 -nitro-o(-isopropylcinnamic acid, or 4-amino-3-methoxy- 0'X-isopropylcinnamic acid. 3 The pharmaceutical composition according to Claim ",wherein the active ingredient is the 24-methylene- cycloartanyl ester of p-acetoxycinnamic acid, p-hydroxy- cinnamic acid, m- or p-nitrocinnamic acid, m- or p-amino- cinnamic acid, in-, or p-hydroxybenzoic acid, in-, o-, or p-nitrobenzoic acid, mn-, or p-aininobenzoic acid, m-acetoxybenzoic acid, in- or p-nitro-4C-methylcinnamic acid, C~ m- or p-ainino-cOLmethylcinnamic acid, 3- or 4-propionyloxy- OC-methylcinnamic acid, 3- or 4-hydroxy-C -methylcinnamic acid, 3 -butyryloxy-0(-ethylcinnamic acid, or 3-hydroxy-L- ethylcinnamic acid. 12 57 1 A' The pharmaceutical composition according to Claim 2) wherein the active ingredient is the 24-methylene cycloartanyl ester of 4-acetoxy-3-methoxycinnamic acid, 4 -acetoxy-3-ethoxycinnamic acid, 4-hydroxy-3-methoxycinnamic acid, 3 -ethoxy-4-hydroxycinnamic acid, 3,4-diacetoxycinnamic acid, 3 4 -dihydroxycinnamic acid, 3-methoxy-4-nitrocinnamic acid, 4 -amino-3-methoxycinnamic acid, cinnamic acid, 5 -amino-2-ethoxycinnamic acid, 4-acetoxy- 3-methoxybenzoic acid, 4-hydroxy-3-methoxybenzoic acid, 4-acetoxy-3-ethoxybenzoic acid, 3-ethoxy-4-hydroxybenzoic acid, 3 ,4-diacetoxybenzoic acid, 3 ,4-dihydroxybenzoic acid, 3-methoxy-4-nitrobenzoic acid, 4-amino-3-methoxybenzoic acid, 3 -methoxy- 4 -propionyloxy-a4-methylcinnamic acid, 4 -hydroxy-3-methoxy-o0-methylcinnamic acid, 4-butyryloxy- 3-methoxy-o(-ethylcinnamic acid, 4-hydroxy-3-methoxy-c 4 ethylcinnamic acid, 4-hydroxy-3-methoxy-oU-butylcinnamic acid, 3 -ethoxy-4-hydroxy-o(-methylcinnamic acid, 4'-hydroxy- 3 -propoxy-Q(-propylcinnamic acid, 3-methoxy-4-nitro-a- methylcinnamic acid, 4 -amino-3-methoxy-g-methylcinnamic acid, 5 -nitro-2-propoxy-o--methylcinnamic acid, or 2 -propoxy-o(-methylcinnamic acid. The pharmaceutical composition according to Claim 20 wherein the active ingredient is the cycloartenyl ester of 3 -methoxy-4-propionyloxycinnamic acid, 3-methoxy- 4 -valeryloxy-O'i-propylcinnamic acid, 4-hydroxy-3-methoxy- 0(-propylcinnamic acid, 4 -capryloxy-3-methoxy-o(--butylcinnamic 258 acid, 3-ethoxy-4-hydroxy-a-ethylcinnamic acid, 3-ethoxy-4-hydroxy-a- butylclnnamic acid, 4-hydroxy-3-propoxy-a-methylcinnamic acid, or 4-hydroxy-3-butoxy-a-methylcinnamic acid.

26. The pharmaceutical composition according to Claim 19, wherein the active ingredient is cyclobranyl ester of 4-hydroxy-3-propoxy-a- ethylcinnamic acid.

27. A pharmaceutical composition for treating hyperlipldemia which comprises a pharmaceutical carrier and an effective amount of cyclobranol as an active Ingredient.

28. A phc-maceutical composition for treatment of hyperlipidemia, substantially as hereinbefore described with reference to any one of the Formulation Examples 1 (excluding Tablets 2 to 4, 6 to 12 or 14 to 21.

29. A triterpenyl ester derived from triterpenyl alcohol and organic acid other than ferulic acid and monobasic and dibasic saturated fatty acids, substantially as hereinbefore described with reference to the Examples. 4t II t

30. A method of treating hyperlipidemia in a patient requiring said treatment, which method comprises administering to said patient an effective amount of a compound according to any one of claims 1 to 10 or 29 and/or a pharmaceutical composition according to claims 17 to 28.

31. A process for producing a triterpenyl ester derived from o triterpenyl alcohol and organic acid other than ferulic acid and monobasic and dibasic saturated fatty acids, substantially as hereinbefore described with reference to the Examples. S "rr DATED this EIGHTEENTH day of APRIL 1990 Amano Pharmaceutical Co Ltd Q F Patent Attorneys for the Applicant SPRUSON FERGUSON TMR/866c'