Howz it Work?: Ipriflavone/Methoxyflavone by Bryan Haycock

[pogue]

I am reposting this from an article originally written for Muscle Monthly Magazine, which is now defunct. Since the article is no longer online, this is the copy from the Google Cache. I am posting this to link within a separate article I am working on currently.

Howz it Work?: Ipriflavone/Methoxyflavone

October 15, 2000

by Bryan Haycock MSc., CSCS

There are a greater variety of supplements available to consumers today than ever before. This is both a blessing and a curse. There are many different products to choose from but at the same time there is a great deal of confusion when it comes to choosing which supplement is right for you. Sure, you can take the easy route and base your purchasing decisions on what the advertisers are telling you, or you can educate yourself and make the wisest and most cost effective purchases for your particular situation. Howz it Work came about as a means of educating people like yourself about how all of these different supplements are supposed to work. If you are serious about your health and performance, you are undoubtedly serious about your supplements. Howz it Work is your key to saving both time and money on your way to achieving success through cutting edge supplementation.

First came Ipriflavone, sold as an anabolic isoflavone. It was preceded by hype and mystery, both tantalizing and frustrating to anybody "in the know" and not on drugs. A lot of people bought it, including me. A lot of people stopped buying it a short time after. A short time after they didn’t notice any new muscle growth, that is. Then came 5-methyl-7-methoxy-isoflavone, a slightly tweaked version of Ipriflavone. It was also preceded by a lot of hype and mystery. As you might expect, a lot of people bought it (not me). It appears people are still buying it.

I have always been one to do my homework on any new supplement I was interested in trying. Not that I’m more savvy than other people, it’s just that I’ve always had a fairly limited supplement budget and couldn’t afford to waste my money. When doing your homework on supplements, you have to ask yourself a couple of questions. First, what is being claimed about a particular supplement? Second, do the claims make sense given the mechanism of action? In trying to answer these two questions you will be lead to the right information to help you make savvy supplement purchases.

So what about Ipriflavone and/or 5-methyl-7-methoxy-isoflavone? What are the claims? What is the mechanism of action?

The claims made by those selling the stuff are as follows:

"… the ultimate, "perfect" anabolic agent. You may be saying to yourself, "I've heard this story before." Well, you may have, but this time the story has a different ending — it's true!"

"It [5-methyl-7-methoxy-isoflavone] was originally designed to increase lean mass on animals and humans, without the negatives associated with steroids."

"…potent anabolic…"

"it's [5-methyl-7-methoxy-isoflavone] not only anabolic, it's healthy!"

"it [5-methyl-7-methoxy-isoflavone] significantly increases calcium, phosphorous, potassium, and nitrogen retention, which clearly shows its anabolic horsepower."

"so powerful it’s guaranteed to help you pack on up to 10 pounds of rock-hard mass in just 30 days!"

"it [5-methyl-7-methoxy-isoflavone] works."

"…5-methyl-7-methoxy isoflavone (methoxivone), the strongest Nutrient partitioning isoflavone. Potent for gaining muscle, losing fat, and increasing vitality, well-being and endurance. Also useful in suppor[t]ing strong, healthy bones, and maintaining low cholesterol levels"

I could go on and on, but I’ll spare you. The point I’m trying to make is that the people making and selling this stuff are telling us it is the "perfect anabolic." That’s a pretty hefty claim!

Our second step in determining the value of ipriflavone/methoxyflavone is to figure out the mechanism of action. The mechanism of action refers to what and how it changes the chemistry of the body. Let’s review the relevant research. Keep in mind that ipriflavone and 5-methyl-7-methoxy-isoflavone (methoxyflavone) are assumed to have the same mechanism of action, so I will use Ipriflavone and methoxyflavone interchangeably.

Ipriflavone inhibits bone resorption:

Most people familiar with Ipriflavone know that it has been extensively studied with respect to bone metabolism, or more specifically, osteoporosis. For instance, one representative animal study found ipriflavone inhibited parathyroid hormone-, vitamin D-, PGE2-, and interleukin 1ß-stimulated bone resorption (1). The effects of Ipriflavone on bone loss have also been demonstrated in humans. The same protocol was used throughout most of these studies (i.e. 200 mg ipriflavone or placebo three times daily). Several two-year studies looked at women immediately postmenopause (age 50-65) and found bone mass was maintained or improved slightly in the ipriflavone groups while those in the placebo groups experienced significant bone loss (2,3,4,5).

This research on bone metabolism doesn’t really back up "the perfect anabolic" claims made by the supplement companies. There still may be hope however, as other research has been done looking at slightly different physiological effects.

Ipriflavone acts as an estrogen "sensitizer" in several tissues:

Before you shriek at the thought of enhancing estrogen’s effects, read on to see exactly how this happens. While Ipriflavone does not have any significant direct estrogenic effects, it does enhance the effects of estrogen, particularly in bone, the thyroid gland, and the gastrointestinal tract (6,7). In bone, ipriflavone makes estrogen’s bone sparing effect more potent. Ipriflavone, like the soy flavone genistein, has decent affinity for the estrogen receptor-beta (8).

OK, so this still doesn’t exactly sound like the "perfect anabolic." Ipriflavone and methoxy do have other effects.

Ipriflavone’s effects on Ca2+:

In the heart, ipriflavone has been shown to prevent Ca2+ ions from building up in the mitochondria (9). Ca2+ builds up in mitochondria when there is insufficient oxygen. Ipriflavone actually enables heart muscle tissue to survive longer without oxygen. This means less damage due to hypoxia (lack of oxygen). No studies have been performed looking at skeletal muscle function during ipriflavone supplementation however, it is not unreasonable to assume a similar function in skeletal muscle. More research is needed in this area.

OK, I will let you in on a little secret. All of the claims about the "anabolic" properties of ipriflavone and methoxyflavone are based on the claims made in their respective patents. Here is an excerpt from one of the patents on Ipriflavone United States patent number 3,949,085.

"Test of anabolic effect: The investigation was carried out with castrated rats by means of the musculus levator ani test and vesicula seminalis test. The preparations were administered orally for a period of three weeks. The tests were performed by the method of Eisenberg and Gordan (Eisenberg, E., Gordan, G. S. J.: J. Pharmacol. 99, 38, 1950). In addition to that, also the weight of the prepared diaphragm of the animals was established. According to these tests, the weight of musculus levator ani rose by a Student significance of p 0.01, the weight of vesicula seminalis did not increase while the weight of the prepared diaphragm of the animals increased by a Student significance of p 0.05. On the basis of these results the preparations proved to possess the anabolic activity free from androgen effect."

The patent goes on to relate…

"The weight yield increasing effect induced by doses of 2 g/100 kg of feed was in the various animal species as follows:

8 to 15% in calves

7 to 10% in cattle

7 to 10% in hogs

8 to 20% in poultry

10 to 20% in rabbits

8 to 12% in guinea pigs"

The amount of feed given to these animals did not increase, only their body weights increased. They did test their new compound on ill humans as well:

"The anabolic effect of the composition was tested on thinned (asthenic), reconvalescent, dystrophic patients suffering from pathological thinness. It has been found that as a result of a treatment lasting for some weeks the patients have gained 2-3 kg of weight. According to our experiments the physical condition of the patients has also improved."

Continued...

[pogue]

In the patent on 5-methyl-7-methoxy-isoflavone (U.S. patent 4,163,746), the details on increasing ipriflavone’s effectiveness are outlined. Several compounds are mentioned, namely 5-methyl-7-methoxy-isoflavone; 5-methyl-7-ethoxy-isoflavone; 5-methyl-7-(2-hydroxy-ethoxy)-isoflavone; and 5-methyl-7-isopropoxy-isoflavone. The same guys filed the 5-methyl-7-methoxy-isoflavone patent some 3 years after they filed the ipriflavone patent. Basically, all this patent does is show that when you methylate the isoflavone, you make it nearly twice as anabolic compared to ipriflavone. The claims are the same except that I noticed the claim that the "compounds are useful as anorexigenic agents. A significant advantage of these compounds over known catabolic agents is that they do not exhibit a central stimulating effect." Now this is quite a claim to add to the fact that they profess that it is an anabolic, as well. So not only does it cause a shift towards more muscle, it also decreases appetite at the same time. Thus far, no manufacturers that I have seen make the claim that ipriflavone or 5-mthyl-7-methoxy-isoflavone is a good appetite suppressants. But if they wanted to I guess they could, based on this patent.

One note about patents; just because an individual or company gets a patent, does not mean that their claims are accurate about their invention. It only means that whoever was assigned their file at the US patent and trademark office approved their file for a patent. It would certainly be nice to see someone do a little research on these flavones to see if in fact there are measurable anabolic effects when given to humans at reasonable doses.

Still, I do feel that ipriflavone/methoxyflavone is a valuable supplement to anybody who trains regularly. It "should" help to strengthen bones and tendons and even enhance cellular metabolism. If the patents are true, it may even increase muscle mass and decrease fat mass if used long enough. Keep in mind that we only answered the first question completely. There is still NO good explanation of how ipriflavone or its methylated derivatives could produce non-hormonal, or hormonal for that matter, anabolic effects.

by Bryan Haycock MSc., CSCS

References:

1. Tsutsumi N, Kawashima K, Nagata H, et al. Effects of KCA-098 on bone metabolism: comparison with those of ipriflavone. Jpn J Pharmacol 1994;65:343-349.

2. Adami S, Bufalino L, Cervetti R, et al. Ipriflavone prevents radial bone loss in postmenopausal women with low bone mass over 2 years. Osteoporos Int 1997;7:119-125.

3. Gennari C, Adami S, Agnusdei D, et al. Effect of chronic treatment with ipriflavone in postmenopausal women with low bone mass. Calcif Tissue Int 1997;61:S19-S22.

4. Agnusdei D, Crepaldi G, Isaia G, et al. A double blind, placebo-controlled trial of ipriflavone for prevention of postmenopausal spinal bone loss. Calcif Tissue Int 1997;61:142-147.

5. Valente M, Bufalino L, Castiglione GN, et al. Effects of 1-year treatment with ipriflavone on bone in postmenopausal women with low bone mass. Calcif Tissue Int 1994;54:377-380.

6. Effect of ipriflavone on the response of uterus and thyroid to estrogen. Life Sci 1986 Feb 24;38(8):757-764.

7. Yamazaki I, Kino****a M. Calcitonin secreting property of ipriflavone in the presence of estrogen. Life Sci 1986;38:1535-1541.

8. Arjmandi BH, Khalil DA, Hollis BW. Ipriflavone, a synthetic phytoestrogen, enhances intestinal calcium transport In vitro. Calcif Tissue Int. 2000 Sep;67(3):225-9.

9. Feuer L, Barath P, Strauss I, Kekes E. Experimental studies on the cardiological effects of ipriflavone on the isolated rabbit heart and in rat and dog. Arzneimittelforschung 1981;31(6):953-8.

Originally Posted on MuscleMonthly.com

[pogue]

United States Patent 3,949,085
Feuer , et al. * April 6, 1976
Anabolic-weight-gain promoting compositions containing isoflavone derivatives and method using same

Abstract

A method of anabolic treatment in humans comprises administering a compound of the following formula: ##SPC1## Wherein R is halosubstituted, nitrosubstituted, or unsubstituted benzyl, R.sup.2 is hydrogen, methyl or carboxy, and R.sup.3 and R.sup.4 are hydrogen, methoxy or nitro. Pharmaceutical compositions are also disclosed.
Inventors: Feuer; Laszlo (Budapest, HU); Nogradi; Mihaly (Budapest, HU); Gottsegen; Agnes (Budapest, HU); Vermes; Borbala (Budapest, HU); Streliszky; Janos (Budapest, HU); Wolfner; Andreas (Budapest, HU); Farkas; Lorant (Budapest, HU); Antus; Sandor (Budapest, HU); Kovaks; Maria (Budapest, HU)
Assignee: Chinoin Gyogyszer-es Vegyeszeti Termakek Gyara RT (Budapest, HU)[*] Notice: The portion of the term of this patent subsequent to September 3, 1991 has been disclaimed.
Appl. No.: 497644
Filed: August 15, 1974
Foreign Application Priority Data
May 27, 1970[HU] 1 996

Current U.S. Class: 514/455
Intern'l Class: A61K 031/35
Field of Search: 424/283
References Cited [Referenced By]
U.S. Patent Documents
3833730 Sep., 1974 Feuer 424/283.

Primary Examiner: Rosen; Sam
Attorney, Agent or Firm: Ross; Karl F., Dubno; Herbert
Parent Case Text


RELATED APPLICATIONS

This application is a division of co-pending application U.S. Ser. No. 371,560 filed 19 June 1973, now U.S. Pat. No. 3,907,830 which in turn is a continuation-in-part of U.S. Ser. No. 146,773 filed 25 May 1971, (now U.S. Pat. No. 3,833,730).
Claims


We claim:

1. An anabolic treatment method for human patients comprising administering to said patients in a daily dosage of 0.2 to 100 mg/kg of body weight a compound having the formula ##SPC11##

wherein R.sub.1 is halosubstituted, nitrosubstituted or unsubstituted propyl, amyl or benzyl, R.sub.2 is hydrogen, methyl or carboxy and R.sub.3 and R.sub.4 are hydrogen or nitro.

2. The method defined in claim 1 wherein said compound is selected from the group which consists of:

7-isopropoxyisoflavone,

7-(4-chlorobenzyloxy)-isoflavone, and

7-isopropoxy-4-nitroisoflavone.

3. The method defined in claim 2 wherein said compound is 7-isopropoxyisoflavone.

4. The method defined in claim 1 wherein said daily dosage is 50 to 1000 mg.

5. The method defined in claim 4 wherein said compound is administered orally in a daily dosage of 300 to 600 mg.

6. An anabolic treatment method for human patients comprising administering to said patients in a daily dosage of 0.2 to 100 mg/kg of body weight a compound having the formula: ##SPC12##

wherein R.sub.1 is halosubstituted, chlorosubstituted or unsubstituted propyl, amyl or benzyl, R.sub.2 is hydrogen, methyl or carboxy, and R.sub.3 and R.sub.4 are hydrogen, methoxy or nitro.

7. An anabolic orally administerable pharmaceutical composition which consists essentially of 0.1 part by weight 7-isopropoxyisoflavone, 0.084 parts by weight potato starch, 0.01 part by weight magnesium sterate and 0.006 part by weight polyvinylpyrrolidine.

8. An anabolic pharmaceutical composition comprising a pharmaceutically acceptable vehicle and an effective amount of a medicament consisting of a compound of the following formula: ##SPC13##

wherein R.sub.1 is halosubstituted, nitrosubstituted or unsubstituted benzyl, R.sub.2 is hydrogen, methyl or carboxy and R.sub.3 and R.sub.4 are hydrogen, methoxy or nitro.

9. An anabolic pharmaceutical composition comprising a pharmaceutically acceptable vehicle and an effective amount of a medicament consisting of a compound of the following formula: ##SPC14##

wherein R.sub.1 is halosubstituted, nitrosubstituted, or unsubstituted propyl, amyl or benzyl; R.sub.2 is hydrogen, methyl or carboxy.

10. An anabolic pharmaceutical composition comprising an effective amount of a pharmaceutically acceptable vehicle and a medicament consisting of a compound of the following formula: ##SPC15##

wherein R.sub.1 is halosubstituted, chlorosubstituted or unsubstituted propyl, amyl or benzyl and R.sub.2 is hydrogen, methyl or carboxy.
Description


FIELD OF THE INVENTION

The present invention relates to metabolic compounds and, more particularly, to anabolic compounds having weight gain promoting effectiveness. The invention also relates to a method of treatment using these compounds and to treatment compositions incorporating same.

BACKGROUND OF THE INVENTION

In order to reduce protein deficiency in nutrition or to terminate it, thorough and significant research is being carried out throughout the world. One of the most obvious ways to achieve this objective is the use of additives to nutrients and feed which improve the utilization of the nutrients introduced into the organism. In animal husbandry, these additives result in a higher body-weight increase for an identical feed consumption and breeding period.

However, it has been rather difficult to develop an appropriate substance for increasing animal body weight since the use of substances with hormonal effects and of antibiotics is not permitted in most of the countries.

Isoflavone compounds have been subjected to a detailed investigation from the aspect of their body-weight increasing effect. These compounds are rather widespread in plants, and a great number of them show oestrogenic properties. (cf. Virtanen, A. J.: Angew. Chem. 30, 544, (1958); Virtanen, A. J., Hietala, P. K.: Acta Chem. Scand. 12, 579, (1958). Grazing animals become infertile on consuming certain varieties of clover; research into this problem has shown that genistein and daidzein present in the plants consumed by pasturing animals are responsible for this effect because of their marked oestrogenic action (Chang, E. W. et al.: Ann. N.Y. Acad. Sci. 61, 625, 1955).

For the determination of the oestrogenic effect of isoflavones a reliable method has been evolved by East, J. (J. Endocrin. 13, 94, 1955). Since that time a number of authors have dealt very thoroughly with this problem (Matrone, G. et al.: Nutrition 59, 235, 1956; Gabor, M.: Naturwise. 461/2 650, 1959; Crabb, P. et al.: J. Am. Chem. Soc. 85, 5258, 1958).

Continued...

[pogue]

The operation can also be performed by the decarboxylation of isoflavone-2-carboxylic acids. Isoflavone-2-carboxylic acids are formed in variant (d) of the process and their decarboxylation may be preferably carried out by heating them in the presence of catalysts such as powdered copper or in the absence of catalysts.

On the basis of the abundant literature on studies concerning the oestrogenic properties of isoflavones and of the data of our research in this field, we were able to produce isoflavone compounds which are devoid of any oestrogenic effect. As described in the above-mentioned application certain types of isoflavone derivatives show a marked weight yield increasing effect which essentially exceeds that of isoflavone compounds possessing definite oestrogenic effects, and this weight yield increasing effect is associated with a reduction or complete absence of oestrogenic effect.

Thus, in the course of our observations in discovering in a field still not described in literature, i.e. in the group nonoestrogenic isoflavones, a very interesting novel biological effect, that of increasing the weight yield, has been found. Beside this action, a significant part of the compounds show anatoxic effects free of androgen effects as well (proved by N-retention and by the musculus levator test or by measuring the skeletal muscle increasing effect). The major part of these compounds were new compounds not synthesized before. It must be emphasized that certain members of this group of compounds show weight-yield increasing effects exceeding all effects of this type known and are at the same time practically in other respects innocuous to the living organism (DL.sub. 50 > g per kg of body weight); the effect is substantially independent of the composition of the feed. The compounds are relatively easily and economically synthesizable on an industrial scale, they are stable, limitlessly storable substances free from taste and odor.

Concerning the effect of the compounds the pharmacological tests given below are of special interest.

Test of anabolic effect: The investigation was carried out with castrated rats by means of the musculus levator ani test and vesicula seminalis test. The preparations were administered orally for a period of three weeks. The tests were performed by the method of Eisenberg and Gordan (Eisenberg, E., Gordan, G. S. J.: J. Pharmacol. 99, 38, 1950). In addition to that, also the weight of the prepared diaphragm of the animals was established. According to these tests, the weight of musculus levator ani rose by a Student significance of p 0.01, the weight of vesicula seminalis did not increase while the weight of the prepared diaphragm of the animals increased by a Student significance of p 0.05. On the basis of these results the preparations proved to possess the anabolic activity free from androgen effect.

During the tests, a total of 30 mg/kg of active ingredient was administered to the animals.

The investigation of nitrogen retention was carried out also with rats. Under a systematic treatment, the nitrogen excretion of the treated animals decreased on the 20th and 30th day, respectively, by a significance of p 0.05.

The result of these investigations similarly pointed to the anabolic effect.

Examinations with S-35 labelled methionine show that under the effect of treatment, increased methionine incorporation takes place in the muscle tissues of the treated animals.

The muscle-activity-increasing effect was investigated by the forced swimming test of rats. The animals were forced to swim in water of 29 deg. C with a load of 3g/100 g body weight. The calorie content and quantity of feed administered was the same as with control animals.

The difference between the periods of forced swimming until exhaustion of the control animals and the animals treated for 45 days and forced to swim daily was 33 minutes (the control animals were subjected to identical treatment with the exception of the active ingredient), i.e. the swimming period (performance) of the control animals increased from 166 to 196 minutes while that of the treated animals from 162 to 225 minutes.

These experiments were carried out by administering daily doses of 5 mg/kg of body weight.

In further experiments we succeeded in partially suppressing the catabolic effect of cortisone with these compounds, and proved that identical doses of anabolic steroids do not exert anabolic effects stronger than that of the invented compounds.

On examining the data of the analysis of body weight it was found that the weight increase of the muscle tissue was specifically greater than that of the fat tissue, and that fat content of the muscle tissue decreased, while that of proteins rose.

The acute toxicity tests proved the full innocuity of the preparation. During a 48 hour period of observation no mice died on administering orally 4000 mg/kg of body weight doses or subcutaneously 3500 mg/kg doses. In rat tests, no perceptible alterations were observed during a 48 hour period, after administering orally or subcutaneously 3500 mg/kg of body weight doses.

On dogs, no alterations were observed during a week of administering 3500 mg/kg body weight doses.

The subacute toxicity tests were performed on rats. When administering daily doses of 200 mg/kg of body weight and 500 mg/kg of body weight orally, no alterations could be observed after one month of test period.

Similar results were obtained in the subacute tests carried out with mice.

As regards chronic toxicity up to the present, after three-month chronic toxicity tests have been completed. After administering daily 100 mg/kg of body weight for three months to male and female rats, no perceivable alterations were observed (full blood investigation, histological and other clinical tests).

Similiarly, negative results were obtained in the toxicity tests with dogs after the first 3 months of observation (here the doses were 20 mg/kg of body weight and 50 mg/kg of body weight).

The oestrogenic effect of the compounds was investigated by the uterus test on infantile mice, after oral and subcutaneous administration. No oestrogenic effects of the compounds were observed.

After administering daily 5 mg/kg of body weight of preparations to chickens for 30 days, the endocrine glands of the experimental animals were subjected to a detailed histological investigation. No perceivable alterations were observed.

The weight yield increasing effect induced by doses of 2 g/100 kg of feed was in the various animal species as follows:

8 to 15% in calves

7 to 10% in cattle

7 to 10% in hogs

8 to 20% in poultry

10 to 20% in rabbits

8 to 12% in guinea pigs

The periods of administration varied from one to four months, depending on the animal species and conditions of breeding. The treated animals did not obtain greater amounts of feed than the control tests during the treatment period. Moreover, in several cases some saving of feed could be attained quite independently of the weight yield increasing effect.

It was observed during the treatment period that the experimentally treated animals showed an increased vitality, and the weight increase was mainly due to an increase of muscle mass. This was particularly evident in pig fattening trials when, in the case of bacon pigs, the ratio of pigs of class A, low in fat, was significantly higher.

In the rats, also the effect exerted on the reproductive organs was separately examined. The capability of reproduction and the number of brood were in case of males and females pretreated with the active ingredient the same as that on untreated controls.

In an investigation on the uptake and excretion of C-14 labelled isoflavones it was found that the uptake is rather quick in both oral and intramuscular administration. After oral administration, half of the ingredient introduced was excreted with urine while the other half with feces.

In a number of organs, activity detectable by radiography was present 48 hours after completion of the treatment.

Continued...

[pogue]

DISCLOSURE OF PARENT APPLICATION

The prior application relates to an animal feed containing as an active ingredient at least one compound of the formula ##SPC2##

Or a salt thereof, in which R.sup.1 represents an unsubstituted or substituted and/or unsaturated alkyl group, R.sup.2 and R.sup.3 represent hydrogen, an alkoxy, nitro, halogen, sulpho or hydroxy group, and R.sup.4 is hydrogen, an alkyl or --COOH group.

The metabolic composition can be favorably applied as a feed additive. In that case one of the compounds of formula I, or thereof, is added to the feed in amounts from 0.00002 to 0.1%, prior to or after the admixture of further additives.

The active ingredient may bear, if desired, also substituents on the alkyl group R.sup.1. Substituents in this position may be a heterocyclic group, a dialkylamino group, carbethoxy, hydroxyalkyl, alkoxyalkyl or aryl or a substituted aryl group. The R.sup.1 alkyl group may be substituted with a nitrogen containing heterocyclic group.

If desired, the compounds of the formula I are mixed with further additives. Substances with biological activity such as vitamins, amino acids, choline chloride, salts of mineral acids, trace elements and other known substances of biological importance are suitable. The feed additive can be used in premixes, in admixture with other components possessing biological effects. As further additives various diluents, solvents, sliding and molding substances, and carriers may be used. The feed additive can be mixed with the feed as a powder, granulate, powder mixture, emulsion or suspension. It is also possible to use the feed composition in mixtures added to the drinking water of the animals.

The earlier application described compounds of the formula ##SPC3##

wherein R.sup.2, R.sup.3 and R.sup.4 have the same definition as in formula I, while R.sup.5 represents an unsaturated or saturated and/or substituted or unsubstituted alkyl group having a carbon chain of more than two carbon atoms and the salts of these compounds. The alkyl group represented by R.sup.5 may bear, if desired, a substitutent such as an aromatic or heteroaromatic ring, dialkylamino groups (such as dimethylamino, diethylamino groups), a carbethoxy group or alkoxy, alkyl or hydroxyalkyl groups. The heterocyclic ring preferably contains a nitrogen atom and an alkoxy, halogen, alkyl or --COOH group as a further substituent.

The new compounds of the formula II can be made by a process wherein ketones of the formula ##SPC4##

a. are reacted with an alkyl orthoformate in the presence of a basic catalyst, or

b. are reacted with hydrogen cyanide and/or cyanides in the presence of hydrogen halide, or

c. are reacted with an alkyl formate in the presence of an alkali metal, or

d. are reacted with an alkyl oxalyl halogenide, followed, if desired, by saponification and/or decarboxylation of the obtained isoflavone ester, or

e. are reacted with an organic anhydride, or

f. are reacted with a N,N-dialkyl acid amide in the presence of phosphorus oxychloride.

g. In an alternative variant, 2-hydroxy-isoflavanone derivatives of the formula ##SPC5##

are dehydrated,

and finally, if necessary, the R.sup.6 group is converted into an R.sup.5 group and/or the compounds are converted into salts or respectively, liberated from their salts;

in the formula

R.sup.2 and R.sup.3 represent hydrogen, a halogen, alkoxy, nitro, sulpho or hydroxy group, R.sup.4 is hydrogen, alkyl or --COOH group, R.sup.5 an unsubstituted or substituted and/or unsaturated or saturated alkyl group with a carbon chain longer than two carbon atoms, R.sup.6 is hydrogen, or a substituted alkyl group or acyl group.

In carrying out variant (a) of the process, the preferred method is to react an appropriately substituted ketone with an orthoformic ester in an aprotic solvent of higher boiling point. Pyridine, dimethyl formamide or diethyleneglycol dimethylether may be used as solvents, while preferably piperidine, morpholine, pyrrolidine and other secondary amines may serve as basic catalysts.

In carrying out variant (b), the preferred method is to react the ketones with hydrogen cyanide, in an aprotic solvent, in the presence of dry gaseous hydrochloric acid or off nonbasic nature, preferably diethylether or other dialkylethers can be used. Zinc chloride or other Lewis acids may be used as catalysts. The reaction is carried out with hydrogen cyanide or with one of its appropriate salts, preferably with zinc cyanide. The mixture may be saturated with dry gaseous hydrochloric acid, and lastly, the formed substituted-form-imino-2-hydroxy-phenyl-benzyl-ketone hydrochlorides are decomposed by treatment with water.

In carrying out variant (c) of the process according to the invention, ketones of the formula ##SPC6##

are reacted with alkyl formates in the presence of an alkali metal. A preferred method is to dissolve an appropriately substituted 2-hydroxy-phenyl-benzyl-ketone in ethyl formate, and adding the solution dropwise to powdered sodium metal, then decomposing the reaction mixture with water, and separating the formed isoflavone.

According to variant (d) of the process, appropriately substituted 2-hydroxy-phenyl-benzyl ketones are reacted with alkyl oxalyl halides. The resulting 2-carbalkoxy-isoflavone derivative can be converted, if desired, into an isoflavone derivative unsubstituted in position 2, by hydrolysis of the ester group followed by decarboxylation. This process can be carried out preferably with methyl or ethyl oxalyl chloride in the presence of a basic acid-binding agent in an appropriate aprotic solvent (preferably pyridine or another tertiary amine capable of binding acids).

According to variant (e) of the process the appropriately substituted 2-hydroxy-phenyl-benzyl ketone is reacted with organic acid anhydrides in the presence of a basic catalyst. The anhydrides of acetic, propionic or benzoic acids can be used as organic acid anhydrides. The anhydride is heated in the presence of a basic catalyst, preferably of the alkali metal salt of the acid component of the anhydride or of a tertiary amine, in the absence of solvents or in an aprotic solvent of higher boiling point such as pyridine or dimethyl formamide.

In carrying out variant (f) of the process according to the invention, the ketone is reacted with N, N-dialkyl acid amides in the presence of phosphorus oxychloride, preferably in a way such that the appropriately substituted 2-hydroxy-phenyl-benzyl ketone is heated with the N, N-dialkyl acid amide (dimethyl formamide, dimethyl acetamide) and phosphorus oxychloride, and using the N, N-dialkyl acid amide (dimethyl acid amide itself as a solvent).

In carrying out variant (g) of the process, 2-hydroxy-isoflavanones of the formula IV are dehydrated by heating alone or in an acidic medium in a polar solvent.

In the course of the process according to the present invention, in the first step, from the compounds of the formula III or IV derivatives can be obtained in which the substituent R.sup.6 is not the R.sup.5 group desired to be present in the end product. In these cases the R.sup.6 group may be converted into and R.sup.5 group. This operation can be carried out by the partial or complete alkylation of mono- and poly-hydroxy-isoflavones, respectively. The alkylation can be preferably performed with alkyl or substituted alkyl halides, alkyl sulfates, olefins and epoxides, preferably in a way such that the alkylating agents are heated with the isoflavones to be alkylated, in appropriate solvents, ketones, dimethyl formamide or ethers having a longer carbon chain, in the case of haloid compounds preferably in the presence of an acid-binding agent such as alkali carbonate, and in the case of alkyl bromides and alkyl chlorides preferably in the presence of an alkali iodide.

The mentioned operation can also be carried out by the partial or complete deacylation and, respectively, the partial or complete dealkylation of acyloxy- and poly-alkyloxy-isoflavones. Acyloxy- and polyacyloxy isoflavones are formed when the procedure of variant (e) is carried out with di- and polyhydroxy-phenyl-benzyl ketones which carry a hydroxy group in the 2 position. Deacylation is preferably performed in an acidic or alkaline medium in the presence of a polar solvent.

Continued...

[pogue]

OBJECTS OF THE INVENTION

It is an object of the present invention to extend the principles originally set forth in application Ser. No. 146,773 now U.S. Pat. No. 3,833,730.

Another object of the invention is to provide nonoestrogenic compounds having anabolic activity and useful in promoting weight gain.

Still another object of the invention is to provide a method of treating human patients to promote weight gain.

Yet another object of the invention is to provide a composition for the treatment of subjects to achieve anabolic activity.

DESCRIPTION OF THE INVENTION

This application deals with compounds within the generic definition of formulas I and II which also conform to the formula ##SPC7##

wherein R.sub.1 is halosubstituted, nitrosubstituted or unsubstituted propyl, amyl or benzyl, R.sub.2 is hydrogen, methyl or carboxy and R.sub.3 and R.sub.4 are the same or different and are hydrogen, methoxy or nitro.

These compounds include compounds in which the group at the 7 position of the isoflavone derivative is:

propyloxy,

amyloxy,

benzyloxy,

chlorobenzyloxy,

nitrobenzyloxy, or

chloropropyloxy;

the group at the 2 position is:

hydrogen,

methyl, or

carboxyl; and

the group at the 3' and 4' positions are:

hydrogen,

methoxy, or

nitro.

Preferably the halo substituent is chloro and the R.sub.3 and R.sub.4 members (at the 3' and 4' positions) are hydrogen or nitro. Where R.sub.1 is propyl, the branched alkyl (isopropyl) is preferred.

The following compounds have been found to be most effective for the present purposes:

7-isopropoxyisoflavone,

7-(4-chlorobenzyloxy)-isoflavone, and

7-isopropoxy-4'-nitroisoflavone.

These compounds, which can be made by the process variants (a) to (g) set forth previously, have the structural formulae: ##SPC8##

7-isopropoxyisoflavone ##SPC9##

7-(4-chlorobenzoxy)-isoflavone ##SPC10##

7-isopropoxy-4'-nitro-isoflavone

The latter compounds have been found to be useful as anabolic agents upon human patients.

For use in human therapy the compounds (individually or in a mixture of two or more) may be processed by the known methods of drug production to form tablets, dragees, powder mixtures, solutions, emulsions or suspensions, primarily for oral administration, in combination with or without vitamin-containing mixtures. The daily dosage is 0.2 to 100 mg/kg of body weight and parenteral or rectal administration may also be used.

Continued...

[pogue]

SPECIFIC EXAMPLE

27 g of 2-hydroxy-4-isopropyloxy-phenyl-benzyl ketone, 22 g of ethyl orthoformate and 5 g of morpholine are boiled for 8 hours in 200 ml of dimethyl formamide. The ethanol formed during the reaction is removed through a fraction head. Then the major part of solvent is distilled off in vacuum and the residue is diluted with dilute aqueous hydrochloric acid. The crude product is filtered and recrystallized from acetone, yielding 24 g of 7-isopropyloxy-isoflavone of m.p. 115.degree.-117.degree.C.

Using a similar method, 7-n-amlyoxy-isoflavone, m.p. 142.degree.-143.degree.C is prepared from 4-n-amyloxy-2-hydroxy-phenyl-benzyl ketone (m.p. 72.degree.-75.degree.C).

EXAMPLE 2

28.6 g of 2-hydroxy-4-isopropyl-phenyl-benzyl ketone or 2-hydroxy-4-n-amyloxy-phenyl-benzyl ketone are dissolved in 50 ml of anydrous ether, 25 g of zinc cyanide are added, and the solution is saturated, under cooling, with dry hydrogen chloride gas. After allowing the mixture to stand for 24 hours, the solvent is decanted from the separated oil, the oil triturated with ether, the ether is decanted, and the residue is heated with 1000 ml of water for 30 minutes on a water bath. The product which precipitates on cooling is filtered, and recrystallized from a mixture of methanol and acetone, thus 7-Isopropyloxy-isoflavone and 7-n-amyloxy-isoflavone, already described in Example 1, is produced.

EXAMPLE 3

A solution of 18 g of 2-hydroxy-4-isopropyloxy-phenyl-benzyl ketone in 150 g of ethyl formate is added in small portions under cooling to 9 g of powdered sodium. After allowing the reaction mixture to stand for some hours, it is treated with ice water containing hydrochloric acid, the ethyl formate is distilled off, the residual aqueous mixture is boiled for an hour, and the product precipitated on cooling is recrystallized from acetone, thus 11 g of 7-isopropyloxy-isoflavone are obtained, m.p. 115.degree.-117.degree.C. In a similar way, 7-n-amyloxy-isoflavone already described in Example 1 can also be produced.

EXAMPLE 4

To a solution of 13.5 g of 2-hydroxy-4-isopropyloxy-phenyl-benzyl ketone in 120 ml of pyridine, 11 ml of ethyloxyalyl chloride are added under cooling. After allowing the reaction mixture to stand for a day, it is diluted with water, extracted with chloroform and repeatedly shaken with a 10% aqueous hydrochloric acid solution. On evaporating the solution, the residue is treated for 5 hours with a mixture of 100 ml of methanol and 50 ml of a 10% aqueous solution of sodium hydroxide, the methanol is distilled off, and the aqueous solution is acidified. The product is filtered, thoroughly dried and, after addition of 5 g of powdered copper, heated to 250.degree.C. On completion of the evolution of gas, the residue is crystallized from methanol, yielding 5 g of 7-isopropyloxy-isoflavone, m.p. 116.degree.-117.degree.C.

EXAMPLE 5

28.6 g of 2-hydroxy-4-isopropyloxy-phenyl-benzyl ketone or 2-hydroxy-4-n-amyloxy-phenyl-benzyl ketone and 25 g of anhydrous sodium acetate are boiled for 14 hours with 120 ml of acetic anhydride under a reflux condenser. The reaction mixture is poured into water, allowed to stand for and the precipitated substance is recrystallized from a mixture of methanol and acetone, yielding 7-isopropyloxy-2-methyl-isoflavone, m.p. 152.degree.-154.degree.C and 7-n-amyloxy-2-methyl-isoflavone, m.p. 87.degree.-89.degree.C.

EXAMPLE 6

16 g of phosphorus oxychloride are mixed with 50 ml of dimethyl formamide with cooling. After 15 minutes, 27 g of 2-hydroxy-4-isopropyloxy-phenyl-benzyl ketone are added, and the mixture is boiled for 18 hours under a reflux condenser. On dilution with water, the precipitate is filtered dried, boiled with 200 ml of methanol, and the methanolic extract is evaporated to a small volume. On recrystallizing the separated crude product from acetone, 10 g of 7-isopropyloxy-isoflavone described in Example 1 are obtained.

EXAMPLE 7

23.8 g of 7-hydroxy-isoflavone in 200 ml of anhydrous acetone are boiled, under stirring, with 18 g of n-amyl bromide, 18 g of potassium carbonate and 1 g of potassium iodide for 72 hours under reflux condenser. The inorganic salts are removed by filtration, the filtrate is subjected to steam distillation in order to remove acetone and excess reagent, the precipitate is filtered and recrystallized from acetone, yielding 7-n-amyloxy-isoflavone, m.p. 120.degree.-122.degree.C. In s similar way, also 7-isopropyloxy-isoflavone, 7-isopropyloxy-2-methyl-isoflavone, and 7-n-amyloxy-2 methyl-isoflavone, and

7-n-propyloxy-isoflavone, m.p. 162.degree.-164.degree.C,

7-n-propyloxy-2-methyl-isoflavone, m.p. 120.degree.-122.degree.C,

7-benzyloxy-2-methyl-isoflavone, m.p. 139.degree.-141.degree.C,

7-(4-chlorobenzyloxy)-isoflavone, m.p. 182.degree.-184.degree.C,

7-(4-chlorobenzyloxy)-2-methyl-isoflavone, m.p. 154.degree.-156.degree.C,

7-(4-nitrobenzyloxy)-2-methyl-isoflavone, m.p. 201.degree.-203.degree.C, and

7-(3-chloropropyloxy)-isoflavone, m.p. 137.degree.-138.degree.C,

can be prepared.

EXAMPLE 8

12 g of 7-hydroxy-isoflavone are boiled for 2 hours under a reflux condenser with 10 g of potassium carbonate and 9 g of isopropyl bromide in 40 ml of dimethyl formamide. On pouring the reaction mixture into water, the separated product is recrystallized from acetone, yielding 7-isopropoxy-isoflavone. In a similar way all the other isoflavone derivatives described in Example 7 can be prepared.

EXAMPLE 9

10 g of 7-hydroxy-2-methyl-isoflavone, 10 g of anhydrous potassium carbonate, 1 g of potassium iddide and 12.5 g of benzyl chloride are boiled in 200 ml of anhydrous acetone for 2 hours with stirring, under a reflux condenser. On subjecting the mixture to steam distillation, the crude product precipitating from a mixture of 100 ml of methanol and 40 ml of acetone, yielding white needle crystalls of 7-benzyloxy-2-methyl-isoflavone, m.p. 139.degree.-141.degree.C.

EXAMPLE 10

10.5 g of 7-hydroxy-isoflavone in 200 ml of anhydrous acetone are boiled for 2 hours with 11.8 g of p-nitrobenzyl iodide in the presence of 5.7 g of anydrous potassium carbonate under a reflux condenser. On distilling off about half of the volume of acetone, the residue is poured into 1000 ml of water. The precipitating crude product is subjected to suction and recrystallized from glacial acetic acid, affording light yellow plates of 7-p-nitrobenzyloxy-isoflavone, m.p. 225.degree.-226.degree.C.

EXAMPLE 11

The daily dosage of 7-isopropoxy-isoflavone and the compounds of formulas VI and VII may vary within wide ranges and depends on the circumstances of the particular case. If administered orally (to humans) the average daily dose may generally amount to 50-1000 mg, preferably 300-600 mg. A preferred dosage consists of 3.times.50 to 3.times.200 mg per day. A composition suitable for oral administration in tablet form has the following composition:

per tablet
______________________________________
7-isopropoxy-isoflavone
0.1 g
Potato starch (amylum solani)
0.084 g
Magnesium stearate 0.01 g
Polyvinylpyrrolidine 0.006 g
Total 0.200 g
______________________________________



The tablets are stable at a temperature of 40.degree., 50.degree. and 55.degree.C. The disintegration time is 6-8 minutes. Another tablet composition is as follows:

per tablet
______________________________________
7-isopropoxy-isoflavone 0.1 g
Avicel (Encompress) 0.1 g
Total 0.2 g
______________________________________



Other anabolic isoflavones may be formulated in an analogous manner.

Continued...

[pogue]

The following test report relates to the anabolic effect of 7-isopropoxy-isoflavone: Detailed experiments on preclinical level were carried out by using 7-isopropoxy-isoflavone. The primary object was the proving of anabolic effect on humans. Experiments were carried out by means of the most up-to-date methods. THe essence of the method was the determination of the nitrogen turn-over.

Principle of the test: After intravenous administration of albumin labelled with I-131 isotope the excretion of labelled albumin was followed with a series of blood sampling. The 50% excretion value -- T.sub.1 /2 -- characteristic of the anabolic effect of the composition. It has been found that the T.sub.1 /2-value has significantly shortened as a result of the administration of the anabolic isoflavone (from 8.03 days to 7.23 days) which is also indicative of the protein incorporation.

The composition has the advantage over steroidal antibiotics that in addition to the shortening of the turn-over period (i.e. increase of the albumin incorporation) it also augments the albumin level within a short time (10-15 days).

In the above test the active compound was administered in 3.times.-150 mg oral doses.

The tests included 10 treated and 10 control patients.

The anabolic effect of the composition was tested on thinned (asthenic), reconvalescent, dystrophic patients suffering from pathological thinness. It has been found that as a result of a treatment lasting for some weeks the patients have gained 2-3 kg of weight. According to our experiments the physical condition of the patients has also improved.

Another characteristic feature of the anabolic effect is the calcium and phosphate retention. These tests were performed on young sheep. It has been found that calcium and phosphorus retention was significantly increased by 20 mg/kg body weight dose.

The essence of the test is as follows: In animals, which were in a state of equilibrium as regards the calcium, phosphate, potassium and nitrogen turn-over, the said complete turn-over was determined prior to and after treatment. The determination of the metal ions was carried out in the fodder and feces by means of flame-photometry after decomposition, while in the urine and drinking water it was performed by means of direct flame-photometry. The inorganic phosphorus was determined, by the method of Fiske-Subbarow and nitrogen was determined according to the micro-Kjeldahl-method.

the calcium retention effect was also proved by means of the isotope method in rats.

After the above tests the composition was tested in osteoporosis of immobilisation and endocrine origin. In this test we also succeeded in proving the calcium retaining effect of the composition in a daily dosage of 300 mg per diet. The subjective complaints, pain-sensation of the patients have considerably decreased.

EOF

[pogue]

End of 3,949,085.

[pogue]

United States Patent 4,163,746
Feuer , et al. August 7, 1979
Metabolic 5-methyl-isoflavone-derivatives, process for the preparation thereof and compositions containing the same

Abstract

Nonestrogenic 5-methyl-alkoxy-isoflavone, namely, 5-methyl-7-methoxy-isoflavone, 5-methyl-7-ethoxy-isoflavone, 5-methyl-7-isopropoxy-isoflavone, and 5-methyl-7-(2-hydroxy-ethoxy)-isoflavone are useful as weight-gain promoters in feeds from which the 7-methoxy and 7-ethoxy isoflavones have been excluded because of estrogenic effects. They are more effective than 7-isopropoxy-isoflavone as well.
Inventors: Feuer; Laszlo (Budapest, HU); Farkas; Lorand (Budapest, HU); Nogradi; Mihaly (Budapest, HU); Vermes; Borbala (Budapest, HU); Gottsegen; Agnes (Budapest, HU); Wolfner; Andras (Budapest, HU)
Assignee: Chinoin Gyogyszer es Vegyeszeti Termekek Gyara Rt. (Budapest, HU)
Appl. No.: 845680
Filed: October 26, 1977
Foreign Application Priority Data
Jul 09, 1973[HU] CI 1396

Current U.S. Class: 549/403
Intern'l Class: C07D 311/02
Field of Search: 260/345.2
References Cited [Referenced By]
U.S. Patent Documents
3352754 Nov., 1967 Gazave 260/345.
3833730 Sep., 1974 Feuer et al. 260/345.
Foreign Patent Documents
2125245 Dec., 1971 DE 260/345.


Other References

Moersch et al., J. Med. Chem., 10, 154, (1967).

Primary Examiner: Chan; Nicky
Attorney, Agent or Firm: Ross; Karl F.
Parent Case Text


CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of application Ser. No. 486,513 filed 8 July 1974, now abandoned.
Claims


We claim:

1. An anabolically effective compound for use as a pharmaceutical or a feed additive of the formula ##STR5## wherein R' is methyl, ethyl, isopropyl or hydroxyethyl; and R.sup.2, R.sup.3 and R.sup.4 are hydrogen.

2. The compound defined in claim 1 which is:

5-methyl-7-methoxy-isoflavone.

3. The compound defined in claim 1 which is:

5-methyl-7-ethoxy-isoflavone.

4. The compound defined in claim 1 which is:

5-methyl-7-ixopropoxy-isoflavone.

5. The compound defined in claim 1 which is:

5-methyl-7-(2-hydroxy-ethoxy)-isoflavone.

Continued...

[pogue]

Description


This invention relates to 5-methyl-isoflavone derivatives suitable for use as or in animal feeds.

According to a feature of the present invention, there are provided new compounds of the formula I ##STR1## wherein

R.sup.1 is hydrogen or substituted or unsubstituted alkyl or substituted or unsubstituted aralkyl;

R.sup.2 and R.sup.3 are each hydrogen, alkyl or alkoxy;

R.sup.4 is hydrogen or alkyl.

Similar isoflavone-derivatives unsubstituted in position 5 are described in U.S. Pat. No. 3,864,362. These compounds possess useful metabolic properties.

If R.sup.1 stand for substituted or unsubstituted alkyl, the alkyl may be a straight or branched chained alkyl group, having 1-20 carbon atoms. The substituent of the alkyl group may be preferably a hydroxy or C.sub.1-16 alkoxy group (e.g. methyl, ethyl, isopropyl, hydroxyethyl, 2-methyl-propyl, 3-methyl-butyl, hexadecyl, etc.).

If R.sup.1 is an optionally substituted aralkyl group, it contains preferably 7-9 carbon atoms (e.g. benzyl, beta-phenyl-ethyl).

R.sup.2, R.sup.3 and R.sup.4 are preferably hydrogen.

Particularly preferred representatives of the compounds of the formula I are the following derivatives:

5-methyl-7-methoxy-isoflavone;

5-methyl-7-(2-hydroxy-ethoxy)-isoflavone;

5-methyl-7-ethoxy-isoflavone; and

5-methyl-7-isopropoxy-isoflavone.

The process for the preparation of compounds of the formula I comprises

(A) REACTING A KETONE OF THE FORMULA II ##STR2## wherein R.sup.1, R.sup.2 and R.sup.3 have the above-stated meanings with a trialkylorthoformiate in the presence of a basic catalyst; or

(B) REACTING A KETONE OF THE FORMULA II with hydrogen cyanide and/or a cyanide salt in the presence of a hydrogen halide; or

(C) REACTING A KETONE OF THE FORMULA II with an alkyl formiate in the presence of an alkali metal; or

(d) reacting a ketone of the formula II with an alkyl-oxalyl-halide and subjecting the isoflavone ester thus obtained to saponification and/or decarboxylation; or

(e) reacting a ketone of the formula II with an organic acid anhydride; or

(f) dehydrating a 7-hydroxy-isoflavanone of the formula III ##STR3## and, if desired, alkylating or aralkylating a compound of the formula I, wherein R.sup.1 is hydrogen.

In carrying out variant (a) of the process, the preferred method is to react an appropriately substituted ketone with an orthoformic ester in an aprotic solvent of higher boiling point.

Pyridine, dimethyl formamide or diethyleneglycol dimethylether are used as solvents, while preferably piperidine, morpholine, pyrrolidine and other secondary amines serve as basic catalysts.

In carrying out variant (b), the preferred method is to react the ketones with hydrogen cyanide, in an aprotic solvent, in the presence of dry gaseous hydrochloric acid or of other hydrogen halides and Lewis acids. In this reaction, also aprotic solvents of non-basic nature, preferably diethylether or other dialkylethers, can be used. Zinc chloride or similarly acting Lewis acids may be used as catalysts. The reaction is carried out with hydrogen cyanide or with one of its appropriate salts, preferably with zinc cyanide. The mixture may be saturated with dry gaseous hydrogen chloride, and lastly, the formed substituted .alpha.-formimino-2-hydroxy-phenyl-benzyl-ketone hydrochlorides are decomposed by treatment with water.

In carrying out variant (c) of the process, ketones of the aforementioned formula II are reacted with alkyl formates in the presence of an alkali metal. A preferred method is to dissolve an appropriately substituted 2-hydroxy-phenyl-benzyl-ketone in ethyl formate, and adding the solution dropwise to powdered sodium metal, then decomposing the reaction mixture with water, and separating the formed isoflavone.

According to variant (d) of the process appropriately substituted 2-hydroxy-phenyl-benzyl-ketones are reacted with alkyl oxalyl halides. The formed 2-carbalkoxy-isoflavone derivative is converted, if desired, into an isoflavone derivative unsubstituted in position 2 by hydrolysis of the ester group followed by decarboxylation. This process is carried out preferably with methyl or ethyl oxalyl chloride in the presence of a basic acid-binding agent in an appropriate aprotic solvent (preferably pyridine or an other tertiary amine capable of binding acids).

According to variant (e) of the process, the appropriately substituted 2-hydroxy-phenyl-benzyl-ketone is reacted with organic acid anhydrides in the presence of a basic catalyst. The anhydrides of acetic, propionic or benzoic acids can be used as organic acid anhydrides. The anhydride is heated in the presence of the basic catalyst, preferably of the alkali metal salt of the acid component of the anhydride or of a tertiary amine, in the absence of solvents or in an aprotic solvent of higher boiling point such as pyridine or dimethyl formamide.

In carrying out of variant (f) of the process, 2-hydroxy-isoflavanones of the formula III are dehydrated by heating alone or in an acidic medium in a polar solvent. Compounds of the formula I, in which R.sup.1 is hydrogen may be subjected to alkylation or aralkylation to provide compounds of the formula I, wherein R.sup.1 is alkyl or aralkyl. The reaction may be carried out by methods known per se. Conventional alkylating agents (e.g. alkyl halides, such as methyl chloride, ethyl iodide, isopropyl bromide, etc. or dialkyl sulphates, e.g. diethyl sulphate) may be used. The reaction may be carried out preferably in the presence of an acid-binding agent (e.g. alkali carbonates, such as potassium carbonate). The reaction is preferably effected in an inert organic solvent, such as acetone or dimethylformamide.

The pharmaceutical compositions comprise, as the active ingredient, a compound of the formula I in admixture with suitable, inert carriers or diluents.

The compounds of the formula I are metabolic agents. Some of these compounds are of anabolic activity. These compounds increase calcium, phosphorous, potassium and nitrogen retention to a significant degree. Due to the above anabolic properties, the compounds are useful in the treatment of osteoporosis of gerontological and immobilation origin. An important advantage of these compounds over anabolic agents belonging to the sterane group is that they do not exhibit androgenic or liver damaging side effects. The decrease of the oxygen consumption of the tissues influences hypoxial or hypercapnial states advantageously in certain cardiological and pulmonological symptoms. The publication of Miklos Gabor, Naturwissenschaften No. 46,650 (1959) describes that isoflavones containing methoxy or hydroxy groups in 5,7,3' and/or 4' positions are estrogenic. The estrogenic activity of the following compounds was specifically disclosed:

Genistein: 5,7,4'-trihydroxyisoflavone;

daidzin: 7,4'-dihydroxyisoflavone;

biochanin: 5,7-dihydroxy-4'-methoxyisoflavone;

formomonetin: 7-hydroxy-4'-methoxyisoflavone;

pratensin: 5,7,3'-trihydroxy-4'-ethoxyisoflavone;

prunetin: 4',5-dihidroxy-7-methoxyisoflavone,

wherein the rings are numbered as follows: ##STR4##

As anabolic agents devoid of side effects, the compounds of the invention are useful in human therapy as roborating agents. They can also be applied in pediatrics.

The following compounds of the formula I are particularly active as anabolic agents:

5-methyl-7-methoxy-isoflavone;

5-methyl-7-ethoxy-isoflavone;

5-methyl-7-(2-hydroxy-ethoxy)-isoflavone; and

5-methyl-7-isopropoxy-isoflavone.

The above compounds are useful as anorexigenic agents. A significant advantage of these compounds over known catabolic agents is that they do not exhibit a central stimulating effect. A further characteristic feature of these compounds is that they reduce the high blood cholesterol level and consequently may be used in the treatment of diseases, being in connection with cholesterinaemia (arterosclerosis, diabetes, etc.).

The pharmaceutical compositions comprise a compound of the formula I and inert solid or liquid carriers. The compositions may be put up in solid (e.g. tablets, capsules, pills) or liquid (e.g. solution, emulsion, suspension) form. The composition contain conventional diluents or carriers (e.g. talc, magnesium stearate, calcium carbonate, starch, water) and optionally further additives (e.g. emulsifying, suspending, wetting agents, etc.). The compositions may also contain further biologically active components (anabolic, appetite-increasing, coronary dilatory agents, vitamin, agents acting on the hear function, etc.). The daily dosage of the compounds of the formula I may vary between wide ranges and depend on the particular conditions of application. A preferred oral dosage unit may be a tablet, pill or dragee, comprising 10-100 mg. of the active ingredient. Two or three tablets may be used a day. The above data are, however, merely of illustrative, rather than restricting character.

Continued...

[pogue]

According to a still further feature of the present invention, there are provided feed additives, comprising at least one anabolic compound of the formula I as the active ingredient.

Particularly useful anabolic feed additives are the following compounds:

5-methyl-7-methoxy-isoflavone;

5-methyl-7ethoxy-isoflavone;

5-methyl-7-(2-hydroxy-ethoxy)-isoflavone; and

5-methyl-7-isopropoxy-isoflavone.

These compounds of the formula I produce a significant weight gain increase in domestic animals as compared to that of the control. It is of importance that the surplus of weight consists of meat, rather than of fat, this is particularly useful in the fattening of pigs. The feed additives of the present invention are useful in the fattening of pigs, cattle and poultry. Thus 5-methyl-7-ethoxy-isoflavone induces a weight-gain of 9-10% on chicken during a fattening period of 5 weeks.

The following test-results show the useful anabolic properties of the compounds of the present invention.

(A) Test animals: broiler, cocks

Test period: 7 weeks.

Number of animals per group: 20

Concentration of test compounds: 2 g./100 kg fodder and 5 g./100 kg fodder.

______________________________________
Weight Amount of fodder
after used for 1 kg. of
Group 6 weeks in g.
body weight (in kg.)
______________________________________
Control 1215 2.75
Compound B
(2 g./100 kg.)
1360 2.57
Compound B
(5 g./100 kg.)
1352 2.50
______________________________________



(B) Test animals sexed cocks:

Test period: 35 days

Number of animals pro group: 30

Concentration of test compound 2 g./100 kg. fodder.

In the first week of the pre-breeding period and the first week of the test period, the animals received a starter fodder, and in the further time of the test a breeding fodder. For 5-12 days the animals were given sulfaquinoxaline as a coccidiostatic agent.

______________________________________
Weight-gain increase in %
Compound related to the control
______________________________________
Compound A + 8.70
Compound B + 6.24
Compound C + 4.33
Compound D + 3.70
Control 0.00
______________________________________



The following test compounds were used:

Compound A=5-methyl-7-methoxy-isoflavone

Compound B=5-methyl-7-ethoxy-isoflavone

Compound C=5-methyl-7-(2-hydroxy-ethoxy)-isoflavone

Compound D=5-methyl-7-isopropoxy-isoflavone.

The starter fodder was of the following composition: Maize 60%; soya (45%) 20%; lucerne meal 2%; fish meal (65%) 10%; yeast 3.3%; calcium phosphate 0.6%; lime 2.3%; sodium chloride 0.3%; vitamin premix I 1.0%; mineral premix I 0.5%.

The composition of the poultry breeding fodder is the following: maize 50%, wheat 14.9%; soya (45%) 12.5%; peanut 9%; lucerne meal 2.0%; fish meal (65%) 4.5%; meat meal (45%) 3.0%; calcium phosphate 1.0%; lime 1.8%; sodium chloride 0.3%; vitamin premix II 0.5%; mineral premix I 0.5%.

The vitamin premixes are of the following compositions:

______________________________________
Compositions of vitamin premixes
Vitamin Vitamin
premix I premix II
0.5% 0.5%
______________________________________
A-vitamin 2,000,000 IU 1,200,000
IU
D.sub.3 -vitamin
400,000 IU 300,000 IU
E-vitamin 4,000 IU 2,000 IU
K.sub.3 -vitamin
400 mg 400 mg
B.sub.1 -vitamin
400 mg 200 mg
B.sub.2 -vitamin
800 mg 700 mg
B.sub.3 -vitamin
1,200 mg 2,000 mg
B.sub.6 -vitamin
400 mg 500 mg
B.sub.12 -vitamin
10 mg 4 mg
Niacine 4,000 mg 5,000 mg
Choline chloride
100,000 mg 100,000 mg
Ethoxy-methyl-quinoline
25,000 mg 25,000 mg
Bacitracin 6,000 mg 4,000 mg
Furazolidone 20,000 mg --
Ardinon -- 25,000 mg
______________________________________


______________________________________
Compositions of mineral premix I
______________________________________
Manganese 20,000 mg
Iron 2,000 mg
Zinc 8,000 mg
Copper 400 mg
Iodine 150 mg
Ethoxy-methyl-quinoline 100 mg
mixed in 100,00 g. of bran.
______________________________________



The feed additives compounds of the formula I are added to the fodder preferably in an amount of 0.0001-0.1%. A preferred active ingredient content is 0.5-5 g./100 kg. fodder, particularly 2 g./100 kg. fodder. The active ingredient content of the fodder may be, however, higher or lower, than the above values.

The compounds of the formula I may be mixed with further additives. Substances with biological activity, such as vitamins, amino acids, choline chloride, salts of mineral acids, trace elements and other known substances of biological importance are suitable. The feed additive can be applied in premixes, in admixture with other components possessing biological effect. As further additives various diluents, solvents, sliding and molding substances and carriers may be used. The feed additive can be mixed to the feed as a powder, granulate, powder mixture, emulsion or suspension. It is also possible to use the feed composition in mixtures added to the drinking water of the animals.

Further details of our invention are to be found in the examples, without limiting the scope of our invention to the examples.

Continued...

[pogue]

EXAMPLE 1

A mixture of 25 g. of 2,4-dihydroxy-6-methyl-phenyl-benzyl-ketone, 5 ml. of morpholine, 25 ml. of triethyl-orthoformiate and 100 ml. of dimethylformamide is refluxed under a fractionating column. The alcohol formed is distilled off. After the distillation of alcohol has ceased, the reaction mixture is heated to boiling for a further period of 30 minutes. The mixture is diluted with water, the precipitated product is filtered off and recrystallized from glacial acetic acid. The melting point of the 7-hydroxy-5-methyl-isoflavone thus obtained amounts to 241.degree.-242.degree. C.

EXAMPLE 2

25 g. of 2,4-dihydroxy-phenyl-benzyl-ketone are dissolved in 500 ml. of ether, 20 g. of zinc-cyanide are added and the solution is saturated with gaseous hydrogen chloride. After standing for 8 hours, the solvent is removed by decanting and the precipitated product is heated to boiling with water for an hour. The precipitated product is filtered off and recrystallized from glacial acetic acid. The 5-methyl-7-hydroxy-isoflavone thus obtained is identical with the product of Example 1.

EXAMPLE 3

A mixture of 13 g. of 5-methyl-7-hydroxy-isoflavone, 75 ml. of dried acetone, 13 g. of anhydrous potassium carbonate and 10.5 ml. of diethylsulphate is refluxed under stirring for 4 hours. Three quarters of the acetone is distilled off and to the residue water is added. The fluocculent product is filtered off recrystallized from methanol. The melting point of the 5-methyl-7ethoxy-isoflavone thus obtained amounts to 112.degree.-114.degree. C. The product froms colorless crystals.

The 5-methyl-7-methoxy-isoflavone is obtained in an analogous manner. Mp.: 117.degree.-119.degree. C.

EXAMPLE 4

13 g. of 5-methyl-7-hydroxy-isoflavone are dissolved in 70 ml. of dimethylformamide, whereupon 26 g. of powdered potassium carbonate and 20 ml. of isopropyl bromide are added and the reaction mixture is stirred at 80.degree. C. for 6 hours. The mixture is cooled, poured into water, the precipitated crude product is filtered off and crystallized from methanol. The melting point of the 5-methyl-7-isopropoxy-isoflavone amounts to 94.degree.-96.degree. C. The product forms colorless needles.

In an analogous manner the following compound is prepared:

5-methyl-7-(2-hydroxy-ethoxy)-isoflavone, mp.: 135.degree.-139.degree. C.

EXAMPLE 5

1 g. of 5-methyl-7-ethoxy-isoflavone and a solution of 1 g. of sodium hydroxide in 20 ml. of 50% aqueous ethanol is heated to boiling for 2 hours. The reaction mixture is cooled, diluted with 20 ml. of water and neutralized with phosphonic acid. In the form of colorless crystals melting at 53.5.degree. C., 2-hydroxy-4-ethoxy-phenyl-benzyl-ketone is obtained.

In an analogous manner the following compounds are prepared:

2-hydroxy-4-methoxy-phenyl-benzyl-ketone, mp.: 90.degree. C.;

2-hydroxy-4-(2-hydroxy-ethoxy)-phenyl-benzyl-ketone, mp.: 117.degree.-119.degree. C.;

2-hydroxy-4-isopropoxy-phenyl-benzyl-ketone, mp.: 58.5.degree.-59.5.degree. C.;

2-hydroxy-4-(2-methyl-propoxy)-phenyl-benzyl-ketone, mp.: 57.degree.-59.degree. C.;

2-hydroxy-4-(3-methyl-butoxy)-phenyl-benzyl-ketone, mp.: 60.degree.-61.5.degree. C.

EXAMPLE 6

Formulation of poultry raising feed:

______________________________________
Maize 40.0 kg.
Feed wheat 20.0 kg.
Bran 6.0 kg.
Extracted Soyabeans 13.0 kg.
Extracted groundnut 11.5 kg.
Powdered alfalfa 1.4 kg.
Extracted sunflower seed 4.0 kg.
Potassium-phosphorus composite (Foszkal)
0.5 kg.
Feed lime 2.3 kg.
Feed sodium chloride 0.3 kg.
Vitamin premix 2 0.5 kg.
Mineral Premix II 0.5 kg.
Total: 100.0 kg.
______________________________________



+2 g. of 5-methyl-7-isopropoxy-isoflavone/100 kg. of feed.

EXAMPLE 7

A fodder of the following compositions is prepared for pigs:

______________________________________
Bran 22.0 kg.
Extracted soyabeans 15.0 kg.
Extracted groundnut 6.0 kg.
Powdered linseed 14.0 kg.
Powdered alfalfa 4.0 kg.
Powdered milk 15.0 kg.
Fish meal 10.0 kg.
Yeast 2.0 kg.
Feed lime 6.0 kg.
Feed sodium chloride 1.5 kg.
Vitamin premix 3.0 kg.
Mineral premix 1.5 kg.
Total: 100.0 kg.
______________________________________



+2 g. of 5-methyl-7-isopropoxy-isoflavone/100 kg. of feed.

As active ingredient other anabolic compounds of the formula I may be used too.

EXAMPLE 8

A tablet suitable for oral administration to humans in the dosage indicated is pressed from the following composition:

______________________________________
Active ingredient of the formula I
0.100 g.
Potato starch 0.084 g.
Magnesium stearate 0.010 g.
Polyvinylpyrrolidine 0.006 g.
Total: 0.200 g.
______________________________________



The tablets are stable at a temperature of 40.degree.-55.degree. C. and desintegration time is 6-8 minutes.

EXAMPLE 9

A tablet may be pressed from the following composition:

______________________________________
Active ingredient of the formula I
0.1 g.
Avicel (Encompress) 0.1 g.
Total: 0.2 g.
______________________________________


______________________________________
Comparative test report
Test animals: one week old cocks of 110 g. in four groups con-
taining 20 animals each
______________________________________
Duration of the Average weights (g)
treatment (weeks)
group 1 group 2 group 3
______________________________________
3 weeks 440 443 446
4 weeks 610 634 580
5 weeks 842 867 742
6 weeks 1013 1097 903
7 weeks 1310 1360 1232
______________________________________
Feedstuff per one kg.
of live weight
2.63 2.57 2.83
______________________________________
For group
2 g. of 7-isopropoxy-isoflavone/q added
1: to feedstuff
For group
2 g. of 5-methyl-7-isopropoxy-isoflavone/
2: q added to feedstuff
For group
1 g. bacitracine/q added to feedstuff
3:
______________________________________
Saving in feedstuff
related to 1 kg. of
Increase in weight
live weight
______________________________________
7-isopropoxy-iso-
flavone 6.3% 8%
5-methyl-7-iso-
propoxy-isoflavone
10.3% 9.2%
______________________________________



Comparative tests

The active compound was incorporated into poultry feed in a concentration of 2 g./100 kg. The test was carried out on broiler chickens which were fed with the feed from an age of 2 days of an age of 5 weeks. In each group 15 chickens were used.

The following test compounds were used:

Compound A=5-methyl-7-methoxy-isoflavone,

Compound B=5-methyl-7-ethoxy-isoflavone,

Compound C=7-isopropoxy-isoflavone.

(i.) The test results are summarized in the following table:

______________________________________
Test Compound Weight increase in %
______________________________________
(A) 8
(B) 8
(C) 4.5
______________________________________



The above results have a significance of p 0.01.

(ii.) The test was carried out as described in (i.) except that the animal groups consisted of 100 broiler chickens. The results are summarized in the following table:

______________________________________
Compound Weight in %
______________________________________
(B) 10
(C) 4.8
______________________________________



* * * * *

EOF