WO1988004931A1 - Method and composition for hair growth in common pattern baldness - Google Patents

Abstract

Composition and method for promoting hair growth and preventing hair loss in mammals exhibiting common baldness pattern. In particular, the composition contains isolated omental lipid material incorporated in formulations including lotions, creams, ointment, gels, sprays and sticks.

Description

METHOD AND COMPOSITION FOR HAIR GROWTH IN COMMON PATTERN BALDNESS

CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of U.S. Patent application Serial No. 948,017, filed December 31, 1986 entitled "Method and Composition for Hair Growth in Common Pattern Baldness", now abandoned.

FIELD OF THE INVENTION

This invention relates to a method and composition for stimulating new hair growth in mammals. In particular, a composition including isolated lipids from mammalian omenta is used to prevent hair loss and stimulate new hair growth in mammals exhibiting common pattern baldness.

BACKGROUND OF THE INVENTION

Common pattern baldness or androgenetic alopecia occurs in both sexes in mammals. Whereas there are historical instances where baldness has been desirable there are many more cases where persons exhibiting traits of baldness have attempted to prevent hair loss or camouflage this condition.

The patent literature contains numerous examples of compositions and claims for promoting hair growth. One of these compositions is applied in cream form and contains principally marrow, such as beef marrow, with additions of bergamot oil, alcohol, oil and sulfur as described in U.S. Patent No. 4,520,012 issued on May 28, 1985 to Alfonsi. The patent teaches that the composition enhances blood circulation in the scalp dermis to nourish the hair, possibly due to the effects of the bergamot oil. It is noted, however, that bergamot oil is known to be a skin irritant and Alfonsi also warns that more than 5% bergamot oil in the composition can actually cause hair loss. (Alfonsi, col. 2, lines 6-12). Another hair growth composition was described in U.S. Patent No. 4,503,047 issued on March 5, 1985 to Banfi. This composition contains horse radish extract in combination with allyl isothiocyanate which is believed to stimulate the forming ability of the hair bulbs. Additional elements of the composition include rhodanide ions, copper ions and sulfur containing amino acids.

A pharmaceutical composition claimed to promote hair growth was patented in U.S. Patent No. 4,139,619 issued on February 13, 1979 to Chidsey. This pharmaceutical composition is known under the generic name Minoxidil and is marketed by Upjohn Company of Kalamazoo, Michigan. The preparation is claimed to increase the rate of terminal hair growth (the broad diameter and colored hair that is readily seen) and to convert vellus hair (fine, colorless hair) to terminal hair. Minoxidil is a potent antihypertensive substance which has been documented in studies to cause hair growth in virtually all patients that were orally treated with the drug for more than one year. de Villez, R. L., Arch. Dermatol. 121: 197-202 (1985). However, within four months of discontinuing the Minoxidil treatment all of the newly grown hairs had fallen out. Id. at 197.

It is further noted that in the study applying Minoxidil to the balding areas of 64 male patients, patients were chosen with hair loss ranging from Hamilton type 3 to type 6. Id at 197.

Patterns of hair loss are classified in the now universally accepted system described in Hamilton, James B. Annals of N.Y. Acad. of Sci. 53: 708-628 (1951). Types I-III represent non balding scalps, types IV-V represent slight balding and types VI-VIII represent horse shoe outlined baldness. Types VII and VIII representing the most advanced stages of baldness were not included in the minoxidil study. Of the 64 study participants, only 3 showed an excellent hair growth result. De Villez, supra at page 199. This study indicates that those subjects having advanced baldness, i.e., the greatest degree of baldness and the longest duration of baldness, were the least response to the treatment.

Additional research in the field of hair growth has investigated the topical application of vitamin A or all-trans-retinoic acid (tretinoin), alone and in combination with 0.5% Minoxidil, to study the effects on hair growth. Bazzano, et al., J. of the Amer. Acad. of Derm. 15: 880-893 (1986). Of the subjects who received tretinoin alone, most of the hair growth was of the lanugo type (prenatal hair in mammals).

Vellus hair growth and only some terminal hair growth on the scalps of individuals with male pattern alopecia was seen with the 5% topical solutions of minoxidil. Id at 882.

In initial studies requested by the inventors, it has been demonstrated that the use of the inventive hair growth composition without the addition of vitamins A, D and E does not decrease the effectiveness of the product in either preventing further hair loss or in promoting new hair growth.

Additionally, it has been demonstrated that the vitamins A, D and E inhibit angiogenesis in CAM assay studies. These results indicate that the topical application of vitamins alone to hair follicles may exert a slightly negative effect in new hair growth.

It has further been demonstrated that the inventive composition minus sodium hyaluronate greatly diminishes the effectiveness of the composition, indicating that the hair growth activity may be the result of a synergistic effect of the omental extract and sodium hyaluronate. It is noted that the foregoing analysis represents only one theory of the activity of the inventive product and is not meant to be limiting to the invention.

Therefore, a nonirritating composition which promotes new growth of terminal hair has been unknown in the art until the present invention. It has been unexpectedly shown that isolated omental lipids in combination with sodium hyaluronate and other ingredients promotes new hair growth in mammals.

The term "new hair growth" refers to new hair grown in balding areas genetically programmed for hair follicles, as opposed to merely lengthening already grown hair. The term also refers to terminal hair, or the hairs of the scalp having the larger diameter and being pigmented, as opposed to vellus hair, or the fine colorless hairs.

It is thus an object of the present invention to provide a method and composition for substantially preventing hair loss in mammals within weeks of topical treatment to balding areas to promote new hair growth in such balding areas.

SUMMARY OF THE INVENTION

Topical application of a composition including omental lipid material and sodium hyaluronate promotes new terminal hair growth, while at the same time preventing further hair loss in the progression of androgenetic alopecia.

Additional ingredients include vitamins A, D and E, silicone, emulsifiers, anti-oxidants, skin fillers, hygroscopic agents and preservatives.

The composition may be applied to the skin in the form of lotions, ointments, gels, sticks, or creams, although lotion form is preferable. The composition applied to balding or bald areas of mammalian skin has been observed to stop further hair loss and to promote new hair growth of terminal hairs to enhance hair distribution. In areas not genetically coded for terminal hair growth such as facial or arm skin areas it is observed that application of the composition did not initiate hair growth.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is a diagrammatic flow chart of the fractionation method of porcine omentum hexane extracts.

Figure 2 is a photograph of the vertex of the head of a test subject exhibiting Hamilton type VII-VIII male pattern baldness after application of the inventive composition designated as Lotion 2-36A, for six weeks according to the protocol of Example I infra.

Figure 3 is a photograph comparing the same area of the same test subject as shown in Figure 1 after application of the inventive composition Lotion 2-36A according to the Example I protocol infra, for an additional two and a half months.

Figure 4 is a second comparison photograph of the same test subject's baldness area as shown in Figures 2 and 3 after 5 months of applying the inventive composition Lotion 2-36A.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The principal ingredient of the inventive composition is isolated omental lipid, preferably extracted from mammals. The omental material may be combined with conventional cosmetic forms for applying the material to the area to be treated, such as ointments, lotions, paste, gels, aerosols, sticks and the like which are known in the art. The preferred embodiment is a lotion form which combines the omental material with emulsifiers, antioxidants, hygroscopic agents, chelating agents, skin fillers., silicon and preservatives. When applied to areas genetically coded for hair growth, the novel composition promotes new hair growth and prevents further hair loss in the progression of alopecia.

Omental material may be extracted as described in Goldsmith, et al.,.J. Amer. Med. Assoc., 252: 2034-36 (1984). A variety of mammalian sources may be used to supply the material, such as porcine, bovine, ovine, or feline omentum.

Omental materials are used in wound healing as described in copending U.S. Patent Application, Serial No. 805,206, filed on December 4, 1985 and entitled "Method for Enhancement of Healing of Epithelial Wounds in Mammals" by Catsimpoolas, et al., herein incorporated by reference.

These omental materials are also used in angiogenesis for myocardial conditions such as myocardial infarcts, angina, vascular or coronary implants and angioplasty as described in copending U.S. Patent Application, Serial No. 811,375, filed on December 20, 1985 entitled "Method for Treatment of Angina and Myocardial Infarctions with Omental Lipids" by Catsimpoolas, et al., herein incorporated by reference.

It is also noted that copending U.S. Patent Application Serial No. 811,894, filed on December 20, 1985 entitled "Method for Healing Bone Damage and Composition" by Lane, et al. shows bone healing use of omental materials, this application is hereby incorporated by reference. (la) Preparation of the Lipid Extract from Omentum by Brittle Grinding and Hexane Extraction.

The preferred method for preparation of omentum for use in the hair growth composition is a brittle grinding technique (also know as a cryogenic technique) combined with direct hexane extraction of the thus obtained omentum powder. Porcine omentum is preferred for use in the composition, however, bovine, feline, or any other type of mammalian omentum may be used.

The porcine omentum is cut or broken into pieces. Freezing the omentum is accomplished by a method described in copending U.S. Patent Application, Serial No. 811,507, filed on December 20, 1985 by Kamarei, et al. entitled "Brittle Grinding and Extraction of Animal and Plant Derived Material"; herein incorporated by reference.

The omental material is frozen to lower its temperature to an empirically critical "Brittleness Temperature" which converts the unbreakable, viscous, and sticky material into an extremely brittle and fragile substance.

Freezing can be accomplished via various methods, including:

1. Air Freezing:

(a) Blast freezing

(b) Fluidized-bed freezing

2. Plate Freezing

3. Liquid-immersion Freezing and

4. Cryogenic Freezing (freezant undergoes a change of state):

(a) Liquid nitrogeon (LN₂), - 196°C (77K)

(b) Subliming carbon dioxide (dry ice), -79°C (194K)

(c) CCl₂F₂ (refrigerant -12), -30°C (243K) Cryogenic freezing with LN₂, among all above methods, is the most desirable method. (1b) Cryogrinding Treatment

Upon obtaining the brittle pieces of tissues, they may be transferred to any size reduction equipment and homogenized (ground) for the desired length of time, i.e., a few minutes at 22,000 RPM. At larger scales, roll mills with both attrition and impact grinding could be used. It is important that the tissue be kept at or below its brittle temperature throughout the grinding process. Grinding below the brittleness temperature is required in order to produce the necessary small particle size. For this purpose one could occasionally add, if needed, quantities of LN₂ to the size-reduction equipment, provided that there is a vent to allow exhaustion of the nitrogen vapor generated during the process.

Upon completion of the process, an extremely fine, and free-flowing (non-sticky) cryoground tissue ("tissue powder") results. The resultant powder obviously may contain some granular lumps as well as fine indiscrete particles.

Parallel to the tremendous increase of surface area (i.e., ratio of surface area to volume), there is also an inherent reduction of density. As an example, density of omentum powder is 0.44 (±5%) g/ml which is almost half of the density of lipids extracted from omentum.

(lc) Cryosieving of Tissue Powder The resultant cryoground tissue powder, while extremely fine, is not "uniform" with regard to size. Consequently, if uniformity in a particular size range is desired for the following steps, tissue powder should be sieved at or below its brittleness temperature. For this purpose, one may use stacked stainless steel standard sieves having standard designations as shown in Table 1.

The best uniform fraction of omentum powder is in the range of 150 to 600 urn (micrometers = um).

a - These standard designations correspond to the values for test sieve apertures recommended by the International Organization for Standardization, Geneva, Switzerland.

b - These sieves are not in the standard series but they have been included because they are in common usage, c - 1000 um = 1 mm.

* Excerpt from Association of Official Analytical Chemists, 14th Ed., A.O.A.C., Arlington, Virginia (1984) The uniform tissue powder can be further processed, transferred, or stored at regular freezing temperature (e.g., -18°C = 0°F). However, even at regular freezing temperature some chemical changes (e.g., oxidation of unsaturated lipids, especially because of tremendous surface area generated by cryogrinding; insolubilization or destabilization of proteins; and degradation of pigments and vitamins and other biomolecules) may slowly occur. Reduction of freezer temperature will cause decline of the rates of the above reactions. Consequently, for longer storage times, it is recommended that the final tissue powder be stored at -40°C, under vacuum or inert gas, and in the dark, (to prevent any possible photo-catalytic reactions). Preliminary evaluation shows that various tissue powders stored under the above conditions for up to 2 months, did not show any physical changes (texture, color, odor, etc.) in the product.

To use the uniform cryoground product, one should desirably "thaw" the tissue powder. Since thawing of non-fluid tissues is inherently slower than freezing, when comparable temperature differentials are employed (due to different thermal properties of ice vs. water). Hence, tissue powders may be subject to damage by chemical or physical (and less microbial or enzymatic) means. In light of these considerations, one skilled in the art will recognize that the thawing process must be carefully considered.

(Id) Direct Hexane Extraction of Omentum Powder 500 g. uniform porcine omentum powder was warmed up to room temperature and extracted with 10 times hexane in a glass blender (22,000 RPM, 30 seconds). The resultant pinkish solution was centrifuged at 2000 RPM for 20 minutes. If centrifugation is performed at room temperature, only a pinkish solid layer precipitates as pellet. However, if centrifugation is done at refrigerated temperature (e.g., 5°C), where solubility power of hexane is reduced, a white layer of lipids will also precipitate on top of the proteinacious pinkish pellet. This layer, however, will easily go into solution if the hexane is warmed up to room temperature or, preferably, to 37°C. Hexane extract was subjected to rotary evaporation under vacuum and at 37°C. Total recovery of direct Ijexane extraction of porcine omentum powder was 361.5 g. (i.e., 72.3%).

(1e) Chloroform/Methanol Extraction of Omentum Powder

500 g. uniform porcine omentum powder was warmed up to room temperature and extracted with 10 times chloroform/ methanol (2:1, v/v) in a glass blender (22,000 RPM, 30 seconds). The solvent extract was centrifuged (2,000 RPM, 20 minutes) and subjected to rotary evaporation (under vacuum, 37°C) until dryness, i.e., neither any solvent condensation occurs, nor any solvent odor is present. A whitish chloroform/methanol fraction (CMFr) weighing 388 g. (i.e., 77.6%) was obtained. The CMFr could be further subjected to a hexane/ethanol fractionation.

(1f) Supercritical CO₂ Extraction of Omentum Powder

1905.4 g. uniform porcine omentum powder was extracted with SC-CO₂ in four trial batches. The solvent to feed ratio of 300 (1 lb./min.) was used. A total of 12 extract fractions were collected from each run. The extractor was maintained at 37°C and 3500 psig, first separator at 40°C and 1500 psig, and second separator at 29-30°C and about 1000 psig, where the bulk of extract was collected. The solvent from the second separator was fed into a knock-out vessel at about 1500 psig and then recycled. The residue in the extractor was in the range of 8-17% of the feed. It was interesting to find out that SC-CO₂ extracts about 83-92% of the feed which is more than chloroform/methanol (with traditional homogenization practice). This would suggest that cryogrinding prior to extraction is indeed effective,

(lg) Mild Thermal Extraction of Omentum Powder In order to thermally extract lipids from omentum or any other fatty tissue, it is necessary to: (1) melt or liquidify lipids, (2) separate melted lipids from the rest of the tissue, and (3) filter the resultant oil or fat from residues. All above steps were combined as "one step" as follows:

1000 g. uniform porcine omentum powder was placed in two stainless steel 150 um (micrometer = um) (#100) sieves. These sieves were stacked on the top of 38 um (#400) sieves and collecting pans. The stacks were placed in an oven with mild heat (70°C.) Upon gradual thawing of omentum powder, lipids became gradually melted while proteinaceous materials became denatured. This resulted in the shrinkage and final collapse of the powdery structure of omentum powder. Melted lipid, however, easily passes through a 150 um sieve and fell as droplets on the 38 um sieve, where it was filtered and separated from small residue particles, and finally clear oil was collected in the collecting pans. The total thermal extraction time was 3 hours, during which occasional mixing occurred. Total recovery was found to be 709.5 g. (i.e., 73.6%). It is notable that above recovery does not cover the compression of the remaining oily materials on the top sieve.

(lh) Analysis of Preferred Porcine Omentum Extract Used In Hair Growth Composition

Two samples of porcine omentum were prepared by the preferred brittle grinding technique followed by hexane extraction as discussed in section (Id) supra. Also see McCluer, R., et al, Lipids, (May 1987) reporting feline omentum extract characterization which may be modified for porcine omentum lipids.

The two porcine hexane extracts (known as HxFrl and HxFr III) were fractionated by a method diagrammed in Figure 1 and described infra.

Ten grams of each hexane preparation (HxFrl and HxFrlll) was partitioned between hexane and 93% ethanol and each phase was washed with fresh opposite phase. The washes were then equilibrated with each other and all like phase were combined as illustrated in Figure 1.

Neutral lipids obtained from the evaporation of the hexane phase were next fractionated on a silica gel column, such as a Unisil column, to remove a large quantity of triglycerides eluted with benzene from minor neutral lipids eluted with a benzene/ethyl/acetate/acetic acid mixture. Fatty acids of the triglycerides were analyzed by gas chromatography (GC) and mass spectrometry (MS) based on methyl esters obtained by alkaline methanolysis.

Polar lipids obtained from the ethanol phase were partitioned according to the Folch method following Folch et al., J. Biol. Chem. 226: 497-509 (1957) and described in copending U.S. patent application, serial no. 811,505 filed on December 20, 1985 by Catsimpoolas et al and entitled "Lipids from Omentum and Methods for Cosmetic Use", herein incorporated by reference (i.e., the fraction was dissolved in chloroform/methanol (2:1, 20 volumes, v/w) and 0.2 volumes of water were added. Phases were thoroughly mixed, and allowed to separate.)

Chloroform, methanol and water were used to obtain upper and lower phase components. The lipids in the upper phase were absorbed onto a C-18 reverse phase cartridge; eluted with methanol, acidic lipids (i.e., gangliosides) and neutral lipids (i.e., complex glycolipids); and separated on a DEAE - sephadex column. The Folch lower phase lipids were run on a Unisil chromatography column and the following three fractions were obtained: (1) minor neutral lipids eluted with chloroform;

(2) glycolipids eluted with acetone and methanol; and

(3) phospholipids eluted with methanol.

Each fraction was dried, weighed and analyzed by thin layer chromotography (TLC).

Unisil Fraction of the Hexane Phase Lipids

Benzene Fraction

The triglyceride fraction eluted by benzene constituted about 97% of the first hexane extract (HXFrl).

Fatty acids of the triglyceride fraction were analyzed by

GC and GC/MS and the percentage distribution of the major components was found to be:

16 : 1 ( 2 % )

16 : 0 ( 24 % )

18 : 2 ( 10 % )

18 : 1 ( 30 % )

18 : 0 ( 16 % )

The remaining ten (10) percent consisted of 14:0 and fatty acids that were present at less than 1% of the total.

Benzene and Ethylacetate Fraction

The minor neutral lipids were further fractionated by preparative thin layer chromatography (TLC) into 13 components. The major components of the minor neutral lipids were free fatty acids, cholesterol and diglycerides in the appropriate ratio of 1:1:0.5, as estimated by the TLC charring. The remaining components were present in trace amounts identified as various diglyceride molecular species determined by direct probe mass spectrometry and fatty acid analysis. Most of the monoglycerides distributed into the ethanol phase were seen in the chloroform fraction but were eluted from the Unisil column. Analysis of Ethanol Phase Lipids

Unisil Fractions of Folch Lower Phase Extraction

Chloroform Fraction

A chloroform fraction of approximately 0.44% by weight of the total HXFrl fraction was obtained and analyzed by TLC. This fraction was found to contain the same neutral lipids found in the hexane layer discussed supra.

Cholesterol and free fatty acids make up about 51% of this fraction; monoglycerides represent 12%; diglycerides represent 4%; and triglycerides represent 26%.

Acetone Fraction

Neutral glycolipids in the acetone fraction were determined by TLC analysis to represent less than .001% of the HxFrl extract.

Approximately 50% of the glycolipids was monoglycosyl ceramide; 30% was diglycosyl ceramide; and 20% was triglycosyl ceramide. The glycolipids in this fraction were only 0.05% of the total fraction weight. Several non-polar components were present in this fraction which represent the majority of the fraction weight.

Methanol Fraction

The phospholipid content of the HxFrl was approximately

0.01%. Two major components of this fraction were identified as phosphatidylcholine (about 80% of the fraction) and sphingomyelin (about 20% of the fraction).

Folch Upper Phase DEAE Fractions

Neutral Fraction

The neutral fraction of the HxFrl extract sample is less than 0.001% of the total HxFrl, and contains globosides and three other polyglycosyl ceramides. The TLC mobilities similar to asialo-GMI, Forssman glycolipid and SSEA-1 antigen. The glycolipids of this neutral fraction represent about 16% by weight. The majority of this percentage is contributed to phospholipid, which is partitioned into the upper phase with the gangliosides and polyglycolyl ceramics.

Acidic Lipid Fraction

Gangliosides represent less than 0.001% of the total HxFrl extract in this fraction as analyzed by TLC. This fraction contains 4 major gangliosides. Approximately 33% of the ganglioside was GM3 , 33% GDla 25% GMl and 8% GD3.

Composition of HxFrlll

The second hexane extract sample (HxFrlll) composition was found to be almost" identical to the composition of the first sample, except a significant amount of volitiles, probably moisture, was found in the initial preparation.

The amount of 18:2 fatty acid in the triglyceride fraction (5%) was decreased and the quantity of the polar lipids (i.e., phospholipids and glycolipids) was almost doubled as determined from fraction weights and TLC analysis. The ganglioside content of HxFrlll was estimated to be 0.001%.

See table 2 infra for a composite of the weights and percentage compositions found during fractionation of the HxFrl and HxFrlll extracts.

(2a) Preparation of the porcine and bovine CMFr For preparation of porcine and bovine Chloroform Methanol Fraction (CMFr), fresh and/or frozen omentum was obtained from a local slaughterhouse. Omenta tissues were washed with minimum amount of distilled water, weighed and cut into 1-2 g pieces under sanitary conditions. Upon pooling and mixing of omenta pieces, they were divided into 500 g portions. Extra portions were kept frozen at -25°c. Each 500 g portion was homogenized with four times Phosphate Buffer Saline (PBS) in a glass blender at 22,000 RPM and for 2 min. the resultant homogenate was centrifuged (4000 RPM, 10 min., 5°c), and the lipid cake was collected (about 98% of omentum weight). Solvent extraction (10 times chloroform: Methanol, 2:1) was carried out on the collected lipid cake in a glass blender (22,000 RPM, 30 s ). The organic solvent containing lipid was centrifuged (2,000 RPM, 20 min., 5°c). Although most of the tissue residue was precipitated as pellet, some finer particles were still suspended after centrifugation. Consequently, attempts were made to collect the solvent phase with minimum tissue residues. The solvent extract was subjected to rotary evaporation (under vacuum, 37 °C) until dryness, i.e., neither any solvent condensation occurs, nor any solvent odor is present. The whitish CMFr weighing 70+2% of the omentum was then collected.

(2b) Preparation of the porcine and bovine HxCMFr The chloroform/methanol fraction is warmed up to 37 °C. Four times 95% EtOH (obtained from absolute ethanol) and six times hexane was added to the CMFr and mixed for 20 minutes. The mixture was then transferred to a separatory funnel, wherein the ethanol and hexane phases were allowed to separate in 20 minutes. Hexane and ethanol phases were separately collected. The ethanol phase was then back washed with 6 times hexane, shaken and allowed to separate. The hexane phase was back washed with 4 times 95% EtOH and the phases allowed to separate within 1 hour. If extra hexane is needed to facilitate the separation, it would be added. The collected ethanol and hexane phases seemed to consist of only one phase. However, upon storage of each phase at refrigerator temperature, (i.e., 4°C), each of the collected phases was separated into two new ethanol and hexane phases which were again collected separately.

The total hexane phase is then subjected to rotary evaporation (under vacuum, 37°c) until a translucent hexane extract of chloroform/methanol fraction is obtained (HxCMFr). The recovery of this phase is about 82±3% of CMFr.

Other organic solvents can be used to extract omentum and the invention is not limited to the specific solvents mentioned above.

(2c) Preparation of Omentum Lipid Extract By Supercritical Gas Extraction

We also note the use of supercritical gas extraction for omentum fractions and factors as described infra and in our co-pending U.S. Patent Application Serial No. 793,622, filed October 31, 1985 and entitled "Supercritical Fluid Extraction of Animal Derived Material" by Kamarei and hereby incorporated by reference.

In this process a supercritical fluid such as SC-CO₂ is preferably used to extract omentum. An SCF has increased solvation power at temperatures above the critical pressure (Pc) and critical temperature (Tc).

Polar materials such as gangliosides remain in the residue while the extract contains the more non-polar or lipid materials such as triglycerides. Temperatures used for example are 38-39 °C and extractor pressure is about 3500 psig. Thus these conditions can avoid extraction using toxic materials, or inefficient extraction or use of expensive and time-consuming extractions and materials.

The following example describes the use of supercritical gas extraction for porcine homogenized omentum and porcine omental CMFr.

APPARATUS

A process development unit (PDU) is used. Briefly, the PDU consists of an extractor and three separators, which are housed in an oven at a predetermined temperature. A supercritical solvent in this case, CO₂, is pumped into the extractor and then flows, sequentially, through separators, and the "knock out" vessel. Pressure is maintained by back pressure regulators. Gas, e.g., CO₂, at approximately normal atmospheric pressure, exits the vent, when the extraction is completed.

2(d) Method for Supercritical Gas

Extraction of Porcine Omentum

Omental samples were melted in a separate, nitrogen purged oven at about 40 C, and then transferred into the extractor.

The vessels were purged with low pressure CO₂, and were then brought to the temperatures and pressures indicated in Tables 2-3. CO₂ was then pumped at a rate of about 0.3 lb/min through the system until a weight of about 200 times the sample weight was pumped. Pressure was bled, and samples in each of the vessels and extractor were removed, weighed, and analyzed.

In these experiments, it was observed that close to, and above the critical point of the CO₂ used, dissolving power increased with an increase in temperature, at constant density and with increased density at constant temperature. For example, a portion of the sample, dissolved in the extractor at 3500 psigs, precipitates out at 1500 psigs in the first separator vessels. Further reductions in pressure/density cause additional fractions to precipitate out, until, at ambient pressure, the supercritical gas contains no dissolved material.

The residues obtained from the extractor were found to be insoluble in CO₂. Polar portions of the material, such as gangliosides, were expected to remain in the fraction, while extract fractions were expected to be rich in neutral, non-polar components. This has been observed to be the case, as the insoluble residue is found to contain polar materials, such as gangliosides, while non-polar materials, such as triglycerides, are found in the extracts .

While a single extractor, 3 separators and collecting vessels, and 1 knock out vessel are used in this embodiment, one skilled in the art will recognize that the number and combinations of each of these is a matter of design choice.

TABLE 3

Material: Porcine Adipose Tissue CMFr Extract

CONDITIONS

Sample Weight Charged: 106.5 gm Supercritical Solvent: CO₂ Solvent Recirculated: 46.9 lbs. Solvent to Feed Ratio: 200/1 Solvent Flow Rate: 0.3 lb. /min.

Extractor Separator Separator Se.parator V5 #1 #2 #3

Temperature 38-39 40 37 33 --

(°C)

Pressure 3500 1500 1300 1100 (psig)

Density 0.87 0.69 0.62 0.26

MATERIAL BALANCE

Total Recovered (grams): 81.6 % Recovery: 76.6

V9 V8 V7 *V6 *V5

Weight (grams) 11.4 58.5 10.0 0.7 1.0

Weight (% of feed) 10.7 54.9 9.4 0.7 0.9

*Comments: V9 very viscous, off-white color

*Washed from vessel with Hexane, evaporated off but may be residual. TABLE 4

Material: Porcine Homogenized Omentum

CONDITIONS Sample Weight Charged: 109.6 gm Supercritical Solvent: CO₂ Solvent Recirculated: 48.3 lbs. Solvent to Feed Ratio: 200/1 Solvent Flow Rate: 0.3 lb. /min.

Extractor Separator Separator Separator V5

# 1 #2 # 3

Temperature 38-39 40 37 34 -- (°C)

Pressure 3500 1500 1300 1100 (psig)

Density 0.87 0.69 0.62 0.26 (gm/cc.)

MATERIAL BALANCE

Total Recovered (grams): 82.3 % Recovery: 75.1

V9 V8 V7 *V6 *V5

Weight 36.7 solids 38.6 1.2? 4.1 1.7

(grams)

Weight 33.5 35.2 1.1 3.7 1.6

*Comments: V9 solids tissue-like

V8 clean white solid melted @ 45 °C *Washed from vessel with Hexane, evaporated off but may be residual. (2e) Use of Detergents for Lipid Isolation Lipids are displaced from homogenized cell membranes, or other complexes involving proteins, by amphipathic detergent molecules which render the proteins "soluble" in aqueous media. The released lipid material is recovered by flotation after centrifugation.

A list of possible detergents is given in Tables 4(a) and 4 (b). These are used in concentrations ranging from 0.1 to 2.0% (w/v) and a pH form 7.0 to 8.0

It is interesting to note that when omentum "powder" (rather than traditional cell breaking method) is used, not only processing and handling of the raw material is much easier for all types of follow-up extractions, but the percent lipid recovery of all above examples is higher than the traditional method of cell breaking (i.e., via PBS).

(3a) Preparation of the Total Lipid Extract from Feline Omentum

The method of Goldsmith et al. J. Amer. Med. Assoc, 252; 2034-36 (1984) was used. Also see co-pending U.S. Application S.N. 642,624 filed August 20, 1984 entitled "Angiogenic Factor and Method for Producing Angiogenesis" by Catsimpoolas, et al., both of the above references being hereby incorporated by reference. In another co-pending application U.S. Serial No. 782,724, filed on October 1, 1985 entitled "Compositions Containing Lipid Molecules with Enhanced Angiogenic Activity" by Catsimpoolas et al., hereby incorporated by reference, various omental extracts are analyzed and their components described. Supercritical gas extraction can also be used to extract omental lipids as described in copending U.S. Application 793,622, filed October 31, 1985 entitled "Supercritical Fluid Extraction of Animal Derived Material" by Kamarei.

The omentum was weighed, spread out on to a plastic surface and cut into approximately 4 cm pieces using surgical scissors. The omental pieces (which range in weight from 30 to 66 gm per cat) were placed in a Waring Blender containing 300 ml of phosphate buffered saline (PBS) precooled to 4°C.

Homogenization was performed for 5 minutes at 20,500 rpm. The resulting homogenate was centrifuged in 250 ml plastic bottles at 1600 g in a refrigerated centrifuge at 4°C for 20 minutes. After centrifugation, three fractions were visible in the bottles, i.e. a pellet, a turbid homogenate and a floating cream colored cake.

For the preparation of the total lipid extract, the cake was removed after decantation of the homogenate and weighed. The blender as well as all the solutions were pre-cooled at 4°C for one hour before use. The cake was homogenized in a Waring blender for 2 min with chloroform/ methanol (2:1, v/v) solvent at room temperature at a ratio of 1 gm cake to 10 ml of solvent. The particulate matter was removed by centrifugation at 2000 g in a clinical centrifuge at room temperature for 10 minutes. The clear supernatant was then subjected to rotary evaporation at 37 °C under vacuum to remove the chloroform- methanol mixture. The fatty material was weighed and is referred to as the "total lipid extract", also known as the lipid extract and/or the chloroform-methanol extract or CME, and the CMFr or chloroform-methanol extracted fraction of the omentum.

(3b) Preparation of Feline Omentum Neutral Lipids Twenty-three grams of total lipid extract, CMFr, were dissolved in 120 ml hexane. Eighty ml of 95% ethanol was added and mixed thoroughly. The phases were allowed to separate, then the lower phase was removed and the upper phase (hexane layer) was reextracted with another 80 ml of 95% ethanol. The lower phases were combined and backwashed with 80 ml hexane. The hexane phases were combined and dried by rotary evaporation. The weight of the neutral lipids (mostly triglycerides) was 22.8 gm, or 98.2% yield. This hexane fraction is also known as the the neutral lipid fraction from omentum or the HxCMFr (hexane phase material from the 95% Ethanol Hexane partition of the CMFr). Other hydrocarbons (e.g., pentane, cyclohexane, cyclopentane, benzene, etc.) can be used instead of hexane for the above separation. Methanol, n-propanol, and acetoni-trile can be used instead of ethanol for the above separation. Other methods can also be used as seen and obvious to those skilled in the art in Kates, M. (1972) "Techniques of Lipidology: Isolation, Analysis, and Identification of Lipids", (North-Holland Elsevier P.269-610) hereby incorporated by reference. These methods among others may include acetone precipitation, solvent partition using countercurrent distribution, column chromatography, and/or high pressure liquid chromatography.

A N₂ atmosphere for example can be used to prevent oxidation during extraction.

(3c) Analysis of Fatty Acids from Feline Omentum Neutral Lipids

For this analysis, the neutral lipid fraction was obtained by applying 200 mg of a total omentum lipid extract (CMFr) dissolved in chloroform to a 10 gm silica gel column and eluting with 25 column volumes of chloroform. This fraction was analyzed for fatty acid content by gas chromatography-mass spectrometry (GC-MS). In order to liberate the fatty acids and form their methyl esters an aliquot (2.5 mg) of the neutral lipid fraction was dissolved in 0.67 ml of chloroform and 0.33 ml of 0.5 N methanolic sodium hydroxide was added. The sample was heated at 60°C for one hour, cooled and mixed with 0.2 ml of water to form two phases. The upper phase was removed and discarded. The lower phase was evaporated to dryness with a stream of nitrogen. The fatty acid methyl esters (FAME) were redissolved in n-hexane for analysis by GC-MS. A Finnigan 4500 GC-MS was used to analyse the FAME. Gas chromatography was performed on a 30 M x 0.25 mM methyl silicone bonded phase fused silica capillary column (DB-1, J & W Scientific, Inc.) with helium as the carrier gas at 12 psi. The column oven temperature was programmed from 205°C to 300°C at 2°C/min with the exit end placed directly into the ion source of the mass spectrometer. Mass spectrometry was performed in the electron impact mode (70 eV) with positive ion detection. Spectra were collected and stored at 0.5 sec intervals.

Injection of standard FAMEs demonstrated that under the conditions used, fatty acids with from 12 to 27 carbons could be detected. The sample yielded chromatographic peaks that were identified as 14:0-0.7%, 16:1-0.2%, 16:0-19.5%, 16:1-0.6%, 17.0-1.1%, 18:2-9.8%, 18:1-44.2%, 18:0-24.0% and traces of 15:0 and 17:2. The identities of the FAME peaks were established by comparison of their chromatographic elution properties and mass spectra to those obtained from standard compounds. The percentage composition for each peak (calculated from a chromatogram plotted from the total ion current of each spectra) is of the summed areas of all FAME peaks.

4. Various omental material samples which may also be used in the composition formulation may be prepared as follows:

Reza Sample #1 400 grams of porcine neutral lipids

(HxCMFr, batch #35 and 36-85-P.O.) from omentum powder. This source is obtained by initial extraction which chloroform/methanol followed by removal of polar lipids, via ethanol extraction, from the extract. Reza Sample #2) 300 grams of porcine neutral lipids (HxFr, batch #37-85-P.O.) from omentum powder. This source is obtained directly by hexane extraction of omentum powder.

Reza Sample #3) 300 grams of SuperCritical-CO₂ extract of porcine omentum powder (equal portions of V8 and V9 extracts of the last 3 runs).

Reza Sample #4) 300 grams of Angio-Medical porcine subcutaneous adipose tissue i.e. P.S.A.T. (batch #38-85). This source is directly extracted by hexane (HxFr).

Reza Sample #5) 50 grams of Angio-Medical P.S.A.T. (batch #15-85-F.B.). This source which should be similar to sample #4, is extracted by Supercritical CO₂ (V8).

Reza Sample #6 500 grams of thermally extracted porcine omentum powder (batch #39-85-P.O.).

Formulations of Composition

The hair growth composition may be formulated in any conventional topical form such as lotions, creams, ointments, aerosols, sticks, gels, and other forms known in the art. Preferably, a lotion is formulated having a percentage by weight of mammalian omental material ranging from about 5% to 20% of the topical preparation, preferably from about 15-20%.

The following examples describe the manner and process of making and using the lotion composition and set forth the best mode contemplated by the inventor of carrying out the inventive method, but are not to be construed as limiting. EXAMPLE I

Topical Lotion (2-36A) or Activa^TM Lotion

CTFA NAME TRADE NAME SUPPLIER^* AMOUNT

(%)

Omentum (Porcine HxFr) Angio-Medical 15.00 Retinyl Palmitate and Vitamins A and D Hoffman LaRoche .10 Ergocalciferol Sorbitan Stearate Span 60 ICI 3.05 Propylparaben Propylparaben Mallinckrodt 0.10 Polysorbate 60 Tween 60 ICI 2.2 Glyceryl Stearate Myverol 18-07 Eastman Kodak 1.13 Tocopheryl Acetate Vitamin E Acetate Eisai U.S.A. 1.0 Dimethicone Dow Corning 200, 350 cps Dew Corning 3.0

Carbomer 940 (2% soln.) Carbopol 940 (2% soln. ) B.F. Goodrich 5.0 Water Purified Water USP 48.4

Butylene Glycol 1,3-Butylene Glycol Union Carbide 5.00 Methylparaben Methylparaben Mallinckrodt 0.30 Trisodium EDTA Trisodium EDTA Dow 0.10 Phenoxyethanol Emeressence 1160 Emery 0.50 Triethanolamine Triethanolainine Union Carbide .10 Sodium Hyaluronate Actimoist^R Active Organics 15.00

100.00

* Mallinkrodt-Chemical Co . , P . O . Box 5439 St . Louis , Mo 63147

ICI/Concord Pike/New Murphy Rd . Wilmington DC 19817

Eastman Kodak/343 State ST , /Rochester NY 14650

Eisai USA Inc . /Los Angeles , CA Hoffman LaRoche/

340 Kingsland St/ Nutley , N . J . 07110

Dow Corning/P . O . Box 1767 /Midland , MI 48640

BF Goodrich/ 6100 Oaktree Blvd/Cleveland , Ohio 44131

Union Carbide/Old Ridgebury Rd/Danbury , CT 06817

Dow Chemical Co/ 2020 Dow Centre/Midland Mi 48640

Emery/ 1300 Carew Tower/Cinncinati Ohio 45202

Active Organics/7715 Densmore Ave/Van Nuys Calif 91406 Another lotion formulation is as follows :

LOTION INGREDIENTS (2-17C)

CTFA NAME TRADE NAME SUPPLIER^* AMDUNT (%) Omentum (Porcine HxFr) Angio-Medical 20.00 Sorbitan Stearate Span 60 ICI 4.06 Propylparaben Propylparaben Mallinckrodt 0.10 Polysorbate 60 Tween 60 ICI 2.94 Glyceryl Stearate Myverol 18-07 Eastman Kodak 1.50 Tocopheryl Acetate Vitamin E Acetate Eisai USA 0.20 Dimethicone Dow Corning 200, 350 cps Dow Corning 5.00

Carbomer 940 (2% soln. ) Carbopol 940 (2% soln..) B.F. Goodrich 0.07 Water Purified Water USP 40.16

Butylene Glycol 1,3-Butylene Glycol Union Carbide 5.00 Methylparaben Methylparaben Mallinckrodt 0.30 Trisodium EDTA Triscdium EDTA Dow 0.10 Phenoxyethanol Emeressence 1160 Emery 0.50 Triethanolamine Triethanolamine Union Carbide 0.07 Sodium Hyaluronate Actimoist^R Active Organics 20.00

100.00

*Mallinkrodt-Chemical Co., P.O. Box 5439 St. Louis, Mo 63147

ICI/Concord Pike/New Murphy Rd. Wilmington DC 19817

Eastman Kodak/343 State ST,/Rochester NY 14650

Eisai USA Inc. /Los Angeles, CA Dow Corning/

P.O. Box 1767/Midland, MI 48640

BF Goodrich/6100 Oaktree Blvd/Cleveland, Ohio 44131

Union Carbide/Old Ridgebury Rd/Danbury, CT 06817

Dow Chemical Co/2020 Dow Centre/Midland Mi 48640

Emery/1300 Carew Tower/Cinncinati Ohio 45202

Active Organics/7715 Densmore Ave/Van Nuys Calif 91406 PREFERRED LOTION INGREDIENTS

Sodium Hvaluronate

This ingredient is sold under the trademark

ACTIMOIST^R owned by Active Organics of Van Nuys, California.

The preferred sodium hyaluronate used in the manufacturing of the hair growth composition has the following chemical analysis:

1. molecular weight - not less than 2,500,000;

2. sodium hyaluronate content - not less than 95%;

3. protein - not more than 0.2%;

4. nucleic acid - not more than 0.2;

5. chondroitin sulfate - not detected;

6. moisture content - not more than 6.0%;

7. color and clarity of solution - colorless and transparent.

Vitamin A and Vitamin D

Vitamins A & D , preferably all-trans vitamin A palmitate and vitamin D₃, respectively, are used in a ratio of 1 million units/gm to 200,000 units/gm to equal about 0.10% of the total solution by weight.

MANUFACTURING PROCEDURE FOR LOTION 2-36A

Prior to the manufacturing of the lotion a carbomer 940 solution which is 2% by weight of the composition formula is prepared. To prepare the solution the following ingredients are needed.

Ingredients Amount ( %)

Purified Water 97.7 Methylparaben 0.20

Propylparaben 0.10

Carbomer 940 2.0

Porcine omentum extract prepared by brittle grinding and direct hexane extraction was used in the preparation of this lotion.

To manufacture lotion 2-36A, purified water is added to a main steam-jacketed stainless steel kettle, preferably equipped with an anchor mixer and preferably heated to 80-85°C. Butylene Glycol is then added and mixed. Methylparaben and Trisodium EDTA are sprinkled into the batch and the ingredients are continuously mixed until they are in solution. At that point Carbomer 940 (2% solution) is added. The temperature is maintained at 75 °C and the kettle is covered to avoid evaporation.

In an auxiliary steam-jacketed stainless steel kettle equipped with a propeller mixer, the ingredients of the oil phase are added as follows: Omentum, Sorbitan Stearate, Proplyparaben, Polysorbate 60, Glyceryl Stearate, Tocopheryl Acetate (Vitamin E), Vitamin A, Vitamin D, and Dow Corning 200 Fluid. The mixture is heated to preferably 70 °C while mixing to achieve uniformity of solution. DO NOT OVERHEAT. The oil phase is then added into the main kettle while mixing. Temperatures should be adjusted to 75°C. Triethanolamine, 99% and Phenoxyethanol are added into the combined mixture and the hot water from the jacketed kettle is drained to begin cooling the batch.

The batch is continually mixed and cooled and at approximately 45 °C, sodium hyaluronate is added.

When the mixture has cooled to 35°C, Dow Corning 200. fluid is added. The mixture should be continually mixed and cooled down to approximately 30°C.

At the preferred temperature, remove the batch from the kettle by either gravity or with the aid of a suitable pump into a clean sanitized container. One should avoid incorporating air into the batch while pumping. Once the mixture is contained, the container should be closed tightly to prevent evaporation.

Procedure for Use To use the resulting inventive lotion, it is preferable that the user wash his or her hair once a day, with a mild acting shampoo, such as a baby shampoo. Once the hair is dry, the lotion should be applied to bald and balding areas of the scalp and gently rubbed for approximately ten seconds.

Preferably, lotion 2-36A (lotion I) is applied topically once or twice a day: in the morning after showering and in the evening before bedtime.

Results

Twenty-two (22) healthy male test subjects ranging in age from 27-59 and having Hamilton types of III - VIII were selected to use the inventive Lotion 2-36A at least once a day as described above.

After only three weeks of use, the subjects were interviewed to ascertain if hair loss had been retarded or stopped and whether new hair growth had been promoted. Of the 22 subjects, 8 reported new hair growth. Of the 15 subjects reporting hair loss prior to treatment, 12 persons reported that hair loss had been retarded or stopped. All 22 subjects reported that their hair felt softer and there were no complaints of tackiness or greasiness.

A second group of 25 subjects applied the Lotion 2-36A to balding areas of their scalp at least once a day for more than 2 months. This group consisted of men ranging in age from 23-59 years old having Hamilton types of IV to VIII. Of the 25 subjects, 17 reported new hair growth. Of the 19 subjects reporting hair loss prior to treatment, 15 reported retardation or stoppage after use of the lotion.

One test subject had been using the lotion for almost five (5) months at the time he was interviewed. He is a 56 year old white male classified as having Hamilton type VII-VIII male pattern baldness for 30 years with a positive family history for androgenetic alopecia. Prior to the present topical application of lotion 2-36A, he had had a small hair transplant of approximately 75 grafts to the mid-crown about 20 years ago.

The growth of new terminal hair on this test subject is illustrated in comparison Figures 2-4. At the time Figure 2 was taken, the subject had been using the Lotion 2-36-A for about six weeks. Distinctive new terminal hairs can be seen in the vertex area below the area containing transplanted hairs. Figure 3 includes two photographic views of the test subject's vertex after 2 ½months of applying the Activa TM lotion to the balding area and shows new hair growth. Figure 4 is a photograph taken of the subject's vertex after about five months of treatment illustrating additional new hair, growth. When comparing Figures 2-4, one sees an increase of new hair growth in the bald area of the subject's vertex after only five months of topically applying the Lotion 2-36A to his balding vertex. The overall results of the new hair growth study indicate that approximately 80% of the subjects participating in these studies reported that hair loss had stopped after treatment with the inventive composition for only three weeks. Approximately 32% of the subjects reported new hair growth following treatment for only three weeks.

After two months of treatment, approximately 70% of the 25 test subjects reported new hair growth and approximately 80% of the 19 test subjects reporting hair loss prior to treatment reported that hair loss had been stopped or retarded. Of the 17 test subject reporting new hair growth, 3 subjects had Hamilton types of VII - VIII, while 13 subjects had type V-VI.

These results are significant in view of the fact that studies testing the effectiveness of minoxidil on baldness did not have representation of advanced stages of baldness, namely types VII and VIII. Additionally, even the types V and VI included in the minoxidil study exhibited a low response to the pharmaceutical treatment, de Villez, R.L. Arch Dermatol 121:197-202 (1985).

One test subject liberally applied inventive Lotion 2-36A on one arm and his face two to four times daily for approximately one year without any noticeable hair growth on his balding scalp. When the lotion was applied to bald areas of his scalp, however, he became aware of increased hair density after only one month of local application. No hair growth was observed on the arm and face areas where the lotion had been applied. This result indicates that topical application of the lotion stimulates hair growth in those areas of mammalian skin which are genetically programmed for hair follicles. Further, the lotion has a topical rather than a systemic action.

At least one woman has participated in studies applying Activa^TM lotion to her thinning scalp resulting in a reduction of hair loss and promotion of some new hair growth. EXAMPLE II Studies were conducted by the inventors to demonstrate the effectiveness of the isolated omental material in the hair growth composition and its effect in synergism with other ingredients of the composition. Nine (9) batches of Lotion 2-36A (Activa TM lotion) were made according to the manufacturing procedure supra with the following diversions:

Batch No. Composition

1 Example I manufacturing method, supra

2 Example I manufacturing method, supra

3 Example I manufacturing method supra with the addition of bovine brain gangliosides

4 Example I manufacturing method, supra

5 Example I manufacturing method, supra minus the sodium hyaluronate ingredient

6 Example I manufacturing method, supra minus vitamins A, D and E

7 Example I manufacturing method, supra minus silicon

8 Example I manufacturing method, supra minus the isolated omental lipid material

9 Example I manufacturing method, supra minus polysorbate 60 Fifty-eight (58) healthy male subjects ranging in age from 24-72 and having Hamilton types ranging from II -

VIII were selected and assigned to apply one of the nine batches of Activa TM lotion on bald and balding areas of their scalp once or twice a day according to the protocol of Example I, supra.

The subjects were interviewed from one to four months following application of the Activa TM lotion and results indicate that batch No. 8 lacking the omental lipid material had a significant reduction in new hair growth when compared to new hair growth in subjects applying the preferred composition of the Activa TM lotion (Batch Nos

1, 2, and 4). Further, subjects applying Activa TM lotion minus the sodium hyaluronate (Batch No. 5) also had a significant reduction in new hair growth when compared to new hair growth achieved in subjects applying the preferred Activa

TM composition. Subjects applying Batch No. 6 ( Activa^TM lotion minus vitamins A, D and E) reported a slightly higher percentage of new hair growth than that reported with the use of the preferred Activa^TM lotion.

CAM assay tests were also performed on the nine batches as described in copending U.S. Patent Application,

Serial No. 811,505, filed on December 20, 1985 entitled

"Lipids from Omentum and Methods for Cosmetic Use" by

Catsimpoolas et al; herein incorporated by reference. Results indicate that Activa TM lotion minus omental material (Batch No. 8) did not exhibit positive angiogenic activity and isolated vitamins A, D and E inhibited angiogenesis. These results indicate that new hair growth activity is not dependent upon the presence of the vitamins A, D and E and that it may be due either to the omental lipid content above or to the synergistic effect of the omental material and sodium hyaluronate.

It would be obvious to those skilled in the art that the inventive composition may be made by combining sodium hyaluronate with any type of lipid or fatty material known in the art. These lipids may be isolated from mammalian tissue, as well as plant tissue, according to techniques known in the art.

EXAMPLE III A preferred cream formulation known as Activa^TM Cream is as follows :

CREAM INGREDIENTS ( Activa^TM Cream)

CTFA NAME TRADE NAME SUPPLIER* AMOUNT (%) Omentum (Porcine HxFr) Angio-Medical 25.00 all-trans vitamin A Vitamins A and D Hoffman LaRoche .10

Retinyl Palmitate and

Ergocalciferol

Sorbitan Stearate Span 60 ICI 3.57

Propylparaben Propylparaben Mallinckrodt 0.10

Polysorbate 60 Tween 60 ICI 2.5

Glyceryl Stearate Myverol 18-07 Eastman Kodak 2.15

Tocopheryl Acetate Vitamin E Acetate Eisai USA 1.0

Dimethicone Dew Corning 200, 350 cps Dew Corning 5.00

Carbomer 940 (2% soln.) Carbopol 940 (2% soln.) B.F. Goodrich 12.50

Water Purified Water USP 13.69

Butylene Glycol 1,3-Butylene Glycol Union Carbide 5.00

Methylparaben Methylparaben Mallinckrodt 0.30

Trisodium EDTA Trisodium EDTA Dow 0.10

Phenoxyethanol Emeressence 1160 Emery 0.50

Triethanolamine Triethanolamine Union Carbide .25

Sodium Hyaluronate Actimoist^R Active Organics 25.00

Cetyl alcohol Cetyl alcohol .25

Stearic acid Stearic acid 3.0

100.00 *Mallinkrodt-Chemical Co., P.O. Box 5439 St. Louis, Mo 63147 ICI/Concord Pike/New Murphy Rd. Wilmington DC 19817 Eastman Kodak/343 State ST,/Rochester NY 14650 Eisai USA/Los Angeles CA Hoffman LaRoche/340 Kingsland St/ Nutley, N.J. 07110

Dow Corning/P.O. Box 1767/Midland, MI 48640 BF Goodrich/6100 Oaktree Blvd/Cleveland, Ohio 44131 Union Carbide/Old Ridgebury Rd/Danbury, CT 06817 Dow Chemical Co/2020 Dow Centre/Midland Mi 48640 Emery/1300 Carew Tower/Cinncinati Ohio 45202 Active Organics/7715 Densmore Ave/Van Nuys Calif 91406 This formula variation is manufactured by the process described supra for the lotion formulation. Other formulations of the cream varying the proportions in the cream and achieving a nonseparating cream at 50°C will be obvious to those skilled in the art. There are no therapeutical differences between the topical application of the lotion versus the cream formulations.

EXAMPLE IV

Omentum Cream 2-19c

Another preferred cream formulation is as follows for the inventive composition:

Omentum Cream 2-19C

Deionized Water 19.33

Butylene Glycol 5.00

Methylparaben 0.30

TriSodium EDTA 0.10

2% Carbopol 940 (2% soln.) 12.50

Omentum 25.00

Span 60 3.57

Propylparaben 0.10

Tween 60 2.50

Myverol 18-07 2.15

Tocopheryl Acetate 0.20

Dow Corning 200 Fluid 1.00

Triethanolamine 99% 0.25

Phenoxyethanol 0.50

1% ACTIMOIST ^R Solution 25.00

Dow Corning 200 Fluid, 350 cps 2.50

100.00 The inventive composition is preferably used in lotion form as cream formulations have been found to have a tacky or greasy feeling when applied to scalp areas. However, it may be appreciated that variations of the proportions of formula will be obvious to those skilled in the art. It is further possible to use various lipid omental extracts on fractions and to incorporate the omental material in an ointment, gel, aerosol or stick form.

EXAMPLE V

Irritability, mutagenicity and toxicological studies were run on the inventive composition as described infra. Similarly test methods were described previously in copending U.S. Patent Application, Serial No. 811,505, filed on December 20, 1985 entitled "Lipids from Omentum and Methods for Cosmetic Use" by Catsimpoolas, et al.; herein incorporated by reference.

The Ames mutagenicity test (Ames et al., (1975) Mutation Research 31: 347-364) performed on the porcine omentum extract (POCMFr) by Product Safety Laboratories of 725 Cranbury Rd, East Brunswick, New Jersey 08816. The mutagenic potential of the test material was measured by its ability to induce reverse "(Back") mutations in specially constricted strains of the bacterium Salmonella Typhimuruim. Mutagens which require activation by enzymes found in mammalian tissues are detected in the bacterial assay by the addition of mammalian microsomes (S-9 fractions).

Positive Controls:

In the absence of metabolic activation, N-Methyl-N-nitro-N-nitrosoguanidine (MNNG), 9-amino- acridine (9AA), and 2-nitroflourene (2NF) were used with the appropriate strains. In the presence of metabolic activation, 2-amino-anthracene (2AA) was used as a positive control for all tester strains. Negative Controls:

The spontaneous mutability of each strain in the presence and absence of metabolic activation was determined. Solvent controls were used at the highest volume tested in the experiment.

The Salmonella/mammalian microsome assay of Ames (Ames et al., 1975) utilizes specially constructed strains of the bacterium, Salmonella typhimurium, to detect mutagenic activity of chemicals. The rationale for the test lies in the established correlation between mutagenic activity and carcinogenic potential, and, similarly, with birth defects and heritable disease.

Gene (point) mutations are usually recognized as change in a phenotypic characteristic. In the Ames assay, the dependence of the Salmonella tester strains on an exogenous source of histidine (auxotrophy) is altered by reverse mutation to histidine independence (prototrophy). In this particular system, the reversion occurs by either base-pair substitution or frameshift mutation, depending upon the strain. The various strains carry additional genetic markers which enhance their sensitivity to mutagens.

In the assay, tester strains are exposed to a series of doses of the test article at concentrations up to the highest practicable does within the limitations of toxicity or solubility. Mutagenic activity is manifested by an increase in the numbers of mutant colonies which grow in the absence of histidine, compared to the spontaneous (background) number of mutants. Mutagens which require activation by enzymes found in mammalian tissues, are detected in the bacterial assay by the addition of mammalian microsomes (S-9 fractions). Ames Test Materials: Bacterial Indicator Strains

Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 were originally obtained from Dr. Bruce N. Ames, Berkeley, California. Strains were propagated in nutrient broth containing 0.5% NaCl, checked for appropriate genetic markers, and stored frozen (Revco freezer) after addition of DMSO (dimethyl sulfoxide) to approximately 8% final concentration.

Media

1. Mutrient broth (Difco) was supplemented with 0.5% NaCl.

2. Top agar contained 0.6% Difco agar, 0.5% Nacl, 0.5 mM biotin, and 0.5 mM L-histidine-HCl.

3. Minimal-glucose agar medium contained 1.5% Bacto-Difco agar in Vogel-Bonner Medium E (Vogel and Bonner, 1956) with 2% glucose.

4. Tryptone agar.

Positive Control Chemicals

N-methyl-N-nitro-N-nitrosoguanidine (MNNG), 9-aminoacridine (9-AA), and 2-nitrofluorene (2-NF) were used in the direct mutagenesis assay with indicator strains as shown in Appendix 1.

2-aminoanthracene (2-AA) was used with all strains in the assay with metabolic activation as shown in Appendix 2.

Solvent

Acetone was the solvent used for the test article. 1. S-9 Metabolic Activation Mixture

S-9 was thawed on the day of assay, diluted first in Tris-KCl buffer to a protein concentration of 10 mg/ml, then added to S-9 buffer to give a final concentration of 100 microliter S-9/ml. The complete S-9 mix was passed through a 0.45 um disposable Nalgene filter unit for sterilization and stored at approximately 4°C until use on the same day.

The following components and their final concentrations were used in S-9 buffer:

Component Final Concentration/ml S-9 mix

MgCl₂ 8 micromole

KCl 33 micromole

Component Final Concentration/ml S-9 mix

Glucose-6-phosphate 5 micromole

NADP 4 micromole

NaH₂PO₄ . H₂O, pH 7.4 100 micromole

Methods for Ames Test:

Bacterial Preparation

The cultures used in this assay were started from the frozen permanents in 21 ml of nutrient broth with 0.5% NaCl and incubated 12-16 hours on a shaker water bath at 37°C.

The fresh cultures were stored in a refrigerator while tests for the appropriate genetic markers were conducted. These tests, which were carried out as described by Ames et al. (1975), showed no growth in the absence of histidine, and no growth in the presence of crystal violet or after exposure of ultraviolet light. Strains TA 1535, TA 1537 and TA 1538 were susceptible to ampicillin, while strains Ta 98 and TA 100 were resistant. Preliminary Toxicity Testing

The effect of the test article on the survival of the bacterial strains was determined prior to the Ames bioassay. This was accomplished by adding 0.1 ml of different levels of the test article and solvent to tubes containing 2.0 ml top agar (at 45°C) and 0.1 ml of the tester strain. After mixing, the tube contents were poured onto the surface of tryptone agar plates. The plates were incubated at 37°C for 40-48 hours and the background lawn of bacteria in test article plates was compared to the bacterial lawn in the solvent plates. Toxicity on the tryptone agar plates was detectable by a thinning or disappearance of this background lawn of bacteria.

Plate Assay for the Detection of Direct Acting Mutagens

Assays were performed according to the method of Ames et. al. (1975). The test material was dissolved and diluted in solvent (acetone) on the day of the assay. The following were added, in order, to 2 ml of molten top agar (45°C):

1. 0.1 ml of culture of the appropriate indicator strain;

2. 0.1 ml of the appropriate concentrations of test material, control article, or solvent.

Each tube was prepared individually, immediately vortexed, and the contents poured over the surface of a minimal-glucose agar plate. Each plate was rotated to evenly distribute the top agar before it hardened. All plates were incubated for approximately 48 hours at 37 °C. After incubation, the plates were observed for revertant colonies and the colonies were counted and recorded.

Five doses of test article were tested in triplicate with each strain. Positive control articles for each strain and spontaneous revertant controls consisting of the indicator strains and solvent were run concurrently. The solvent and the highest dose level of the test material were checked for sterility by adding 0.1 ml of each to.2 ml of top agar without the test organisms and pouring the entire contents over minimal-glucose agar plates.

Plate Assay for the Detection of Mutagens Requiring Metabolic Activation

Plate assays with S-9 were conducted as described above, the only modification being the addition of 0.5 ml of S-9 mix to each tube of top agar immediately before vortexing and pouring.

The positive control article for each strain and spontaneous revertant controls consisting of the indicator strains and S-9 mix were run concurrently. The S-9 mix was checked for sterility by adding 0.5 ml to 2 ml of top agar without the test organisms and pouring the entire contents over minimal-glucose agar plates. All plates were incubated approximately 48 hours at 37°C.

Criteria

The following criteria were used in the evaluation and reporting of the mutagenic potential of the test material:

1. The spontaneous revertant levels for each strain when used in either the direct plate assay or the activated plate assay must not differ significantly from the range of historical values shown in Appendix 3.

2. All sterility controls must be negative.

3. All positive controls must demonstrate that the indicator strains are functional with known mutagens as evidenced by an increase of at least three times the number of revertant colonies per plate as the spontaneous revertant controls.

4. To be considered positive for mutagenic activity, the test material should exhibit a dose response effect

(increasing number of revertant colonies with increased amounts of the test sample).

5. To be considered mutagenic for strains TA 1535, TA 1537, TA 1537, TA 1538, and TA 98, the test sample should produce a positive dose response over three concentrations with the lowest increase in revertants/plate greater than or equal to 3x the solvent control value or the S-9 fraction control value, as applicable.

6. To be considered mutagenic for strain TA 100, the test sample should produce a positive dose response over three concentrations with at least one dose producing an increase in revertants/plate greater than or equal to 3.5 x the solvent control value or the S-9 fraction control value, as applicable.

A valid test requires that criteria 1, 2 and 3 be met. To be considered mutagenic, criteria 4, and 5 or 6 must also be met.

AMES TEST

Appendix 1

POSITIVE CONTROL CHEMICALS IN THE DIRECT MUTAGENESIS ASSAY

Positive Indicator Control Cone. Mutagenic

Strain Chemical (ug/plate) Solvent Activity

TA 1535 MNNG 5.0 DMSO Base-pair

Substitution

TA 1537 9-AA 250.0 DMSO Frameshift Mutation

TA 1538 2-NF 0.5 DMSO Frameshift Mutation

TA 98 2-NF 0.5 DMSO Frameshift Mutation

TA 100 MNNG 5.0 DMSO Base-pair

Substitution

MMNG, N-methyl-N-nitro-N-nitrosoguanidine; 9-AA, 9- aminoacridine; 2-NF, 2-nitrofluorene; DMSO, dimethyl- sulfoxide. AMES TEST

Appendix 2

POSITIVE CONTROL CHEMICALS IN THE METABOLIC ACTIVATION MUTAGENESIS ASSAY

Positive Indicator Control Cone. Mutagenic

Strain Chemical (ug/plate) Solvent Activity

TA 1535 2-AA 2.5 DMSO Base-pair

Substitution

TA 1537 2-AA 2.5 DMSO Frameshift Mutation

TA 1538 2-AA 2.5 DMSO Frameshift Mutation

TA 98 2-AA 2.5 DMSO Frameshift Mutation

TA 100 2-AA 2.5 DMSO Base-pair

Substitution

2-AA, 2-aminoanthracene; DMSO, dimethylsulfoxide. AMES TEST

Appendix 3

HISTORICAL BACKGROUND OF SPONTANEOUS REVERTANT LEVELS FOR THE BACTERIAL INDICATOR STRAINS*

Mean Range

(# Revertants Standard (# Revertants Strain /Plate) Deviation /Plates)

A. Plate Assay

Without Metabolic Activation System

TA 1535 18.09 8.04 2-43

TA 1537 6.29 2.47 2-15

TA 1538 13.72 4.48 5-32

TA 98 23.54 9.03 9-60

TA 100 144.94 41.89 66-318

B. Plate Assay With Metabolic Activation System

TA 1535 15.86 5.98 5-40

TA 1537 8.19 2.68 2-15

TA 1538 26.27 10.22 12-65

TA 98 39.85 14.31 16-90

TA 100 156.53 44.61 73-407 EXAMPLE VI

Oral toxicity tests on the PO HxFr (white creme) were done by Product Safety Labs, 340 Commercial Avenue, New Brunswick, NJ 08901. The test material was warmed under tap water until liquified.

During the test period the animals (rats) were each uniquely identified and individually housed in stainless steel wire bottomed cages in an environmentally controlled room with a 12 hour light/dark cycle. Feed and water were provided ad-libitum after dosing.

Procedure

Acute Oral Toxicity. FHSLA, 16 CFR 1500.3. The rats were fasted for 18 hours and then individually and singly dosed by gavage with 5.0 g/kg body weight of test material. The rats were individually caged and observed for mortality or other signs of gross toxicity for 14 days. Feed and water were provided ad-libitum.

The oral LD₅₀ of the Porcine HxFr test material is greater than 5.0 g/kg.

At autopsy, no macroscopic changes could be noted in any of the internal organs of the treated rats and the test material is not toxic as defined in 16 CFR 1500.3.

EXAMPLE VII Oral toxicity tests were also done on the Activa^TM Super Rich Skin Energizer for Day (Lotion) and Activa^TM

Super Rich Skin Energizer (Cream) by Leberco Testing, 123

Hawthorne St., Roselle Park, N.J. 07204.

The test material was administered orally at a dose of 5g/kg to a group of 15 male and 15 females rats as described supra in Example XI.

At autopsy, no macroscopic changes were noted in any of the internal organs of the treated rats and the test material is not toxic as defined in 16 CFR 1500.3. EXAMPLE VIII

Eye irritation studies were done on the PO HxCMFr and the BO HxCMFr (bovine omentum hexane extracted CMFr) on rabbits, by Product Safety Labs of New Brunswick, N.J.

During the test period the six animals were individually housed in wire bottomed cages in an environmentally controlled room with a 12 hour light/dark cycle. Feed and water were provided ad-libitum after dosing.

Procedure

Primary Eye Irritation was done according to FHSLA 16 CFR 1500.42. Six healthy young adult albino rabbits were each uniquely identified. One-tenth of a milliliter (0.1 ml) of the test material was placed on the reverted lower lid of one eye of each rabbit. The upper and lower lids were gently held together for 1 second before releasing, to prevent loss of the test material. The other eye of each rabbit remained untreated and served as a control. Ocular lesions were evaluated by the method of Draize et al. J. Pharmacol. Exp. Ther. 83: 337-390, 1944. The Draize scores were then classified according to Kay and Callandra, J. Soc. Cos. Chem. 13: 281-289, 1962. Lesions were evaluated at 24, 48 and 72 hours.

MMTS* Classification Symbol

0.0 - 0.5 Non-irritating N

0.6 - 2.5 Practically non-irritating PN

2.6 - 15.0 Minimally irritating M₁

15.1 - 25.0 Mildly irritating M₂

25.1 - 50.0 Moderately irritating M₃

50.1 - 80.0 Severely irritating S

80.1 - 100.0 Extremely irritating E

100.0 - 110.0 Maximally irritating M_X * maximum mean total score The MMTS of the PO HxCMFr was 1.00. The material is considered to be practically non-irritating (PN) . The MMTS of the BO HxCMFr was 0.00. The test material is considered to be non-irritating.

EXAMPLE IX

Eye irritation studies were also done on the Activa

TM Super Rich Skin Energizer for Day (lotion) and Activa

TM Super Rich Skin Energizer (cream) on rabbits by Leberco

Testing Inc., 123 Hawthorne St., Roselle Park, N.J. 07204.

Procedure:

Six normal health albino rabbits were used in this test. Each animal has 0.1 ml of the test sample instilled into the right eye with no wash out. The unwashed left eye of each animal served as its own control.

Both the treated and control eyes were examined every 24 hours for three days and observations were recorded according to the "illustrated guide for grading eye irritation by hazardous substances." The test is considered positive if four or more of the animals in the test group exhibited a positive reaction. If only one an-imal exhibits a positive reaction, the test shall be regarded as negative. If two or three animals exhibit a positive reaction, the test is repeated using a different group of six animals.

Neither the lotion nor cream formulations produced a positive reaction and are not considered eye irritants as defined in 16 CFR 1500.42 (a-c).

EXAMPLE X

Prophetic Patch Test

Material is applied to subject's back under the dressing and kept in place for 48 hours. The test site is graded at 15 minutes and 24 hours after the removal of the dressing. Two weeks after the first application a path is left in place for 48 hours, removed and graded 15 minutes and 24 hours later according to the following scale:

SCORING SCALE X Not patched

0 No reaction

0.5 Minimal reaction

1 Definite erythema

2 Erythema with edema

3 Erythema with yesiculation and edema

4 Intense Erythema with bullae G Glazing

5 Scaling

Subjects

100 healthy human adults free of any significant systemic or dermatologic disorder. The area of the back utilized is free of any blemishes which might interfere with grading the test sites. Written informed consent is obtained from are subjects prior to the start of a test.

Prophetic patch test was done on 2 groups of 50 by

Derma-test Laboratories Inc., 29-28 41st Avenue, Long Island City, N.Y. 11101 on the lotion (Activa^TM Super Rich Skin Energizer for Day) and the cream (Activa^TM Super

Rich Skin Energizer). 0.15 ml of the extracts are used.

Also tested was 0.15 ml petrolatum on 102 subjects using

Parke-Davis patch material. In the inducation phase, patches were removed and sites scored 48 and 72 hours after application; all test materials were negative at each reading. Challenge patches were applied 12 days after removal of the inducation patch readings were made at 48 and 72 hours. After application all patch sites were negative at each reading without any. evidence of dermal contact irritation or sensitization. EXAMPLE XI Investigations as to the presence of any viruses have been performed on porcine omentum CMFr in a comprehensive viral. pxofile performed by Damon Clinical Laboratory as also described in copending U.S. Patent Application Serial No. 811,505, filed on December 20, 1985 by Catsimpoolas and herein incorporated.

Claims

What is claimed:

1. Composition comprising isolated omental lipids for topical application to balding human skin for reducing terminal hair loss or promoting new hair growth.

2. Composition of claim 1 wherein the composition promotes new hair growth on human skin genetically programmed for hair growth.

3. Composition of claim 2 wherein new hair growth comprises terminal hair.

4. Composition of claim 1 wherein the omental lipids are derived from the group consisting porcine, bovine, ovine, feline omental lipids and mixtures thereof.

5. Composition of claim 1 wherein the omental lipids are isolated using a cryogenic technique.

6. Composition of claim 5 wherein the omental lipids are isolated by extraction with at least one organic solvent.

7. Copmposition of claim 6 wherein the extraction is a hexane extraction.

8. Composition of claim 6 wherein the omental lipids are isolated with a chloroform-methanol extraction.

9. Composition of claim 5 wherein the omental lipids are isolated by supercritical gas extraction.

10. Composition of claim 9 wherein the gas is CO₂.

11. Composition of claim 1 wherein the omental lipids are isolated by detergent extraction.

12. Composition of claim 1 wherein the omental lipids are isolated using heat extraction.

13. Composition of claim 1 further comprising sodium hyaluronate.

14. Composition of claim 1 further comprising silicone.

15. Composition of claim 1 further comprising hyaluronate and silicone.

16. Composition of claim 1 further comprising emulsifiers.

17. Composition of claim 16 wherein the emulsifiers are selected from the group consisting of glycerol stearate, sorbitan stearate, carbomer, polysorbate 60 and mixtures thereof.

18. Composition of claim 1 further comprising an antioxidant.

19. Composition of claim 18 wherein the antioxidant is selected from the group consisting of EDTA, Vitamin E and mixtures thereof.

20. Composition of claim 1 further comprising Vitamin A and Vitamin D.

21. Composition of claim 1 further comprising preservatives.

22. Composition of claim 21 wherein the preservatives are selected from the group consisting of methylparaben, proplyparaben, phenoxy-ethanol, butylene glycol, EDTA and mixtures thereof.

23. Composition of claim 1 further comprising hygroscopic agents.

24. Composition of claim 23 wherein the hygroscopic agent is selected from the group consisting of silicone compounds, butylene glycol, hyaluronate compounds, water and mixtures thereof.

25. Composition of claim 24 wherein the silicone compounds are selected from the group consisting of dimethicone, diphenicone, phenyldimethicone, cyclomethicone and mixtures thereof.

26. Composition of claim 1 further comprising metal chelating agents.

27. Composition of claim 26 wherein the chelating agent is EDTA.

28. Composition of claim 1 further comprising skin fillers.

29. Composition of claim 28 wherein the skin filler is carbomer.

30. Composition of claim 1 wherein the omental lipid material is incorporated into a lotion, cream, gel, spray, or stick.

31. Composition of claim 1 wherein the composition reduces hair loss in humans having common pattern baldness.

32. Composition of claim 31 wherein the composition promotes new hair growth in humans having common pattern baldness.

33. Composition of claim 32 wherein the composition reduces hair loss or promotes new hair growth in male human subjects having common pattern baldness.

34. Composition of claim 30 wherein the lotion, ointment, cream, gel, spray or stick is a mixture comprising materials selected from group consisting of omental lipid material, sodium hyaluronate, emulsifiers, hygroscopic agents, preservatives, skin filler, metal chelating agents and mixtures thereof.

35. Composition of claim 30 wherein the lotion, cream ointment, gel, spray or stick consists of a mixture comprising materials selected from the group consisting of omental lipid material, sodium hyaluronate, sorbitan stearate, propylparaben, polysorbate 60, glyceryl stearate, tocopheryl acetate, vitamin A, vitamin D, dimethicone, carbomer 940, water, butylene glycol, methylparaben, trisodium EDTA, phenoxyethanol, triethanolamine and mixtures thereof.

36. Process for promoting the rate of new hair growth in humans comprising: topically applying a composition containing isolated omental lipids to' human skin sites having areas genetically programmed for hair follicle growth.

37. Process of claim 34 wherein the new hair growth comprises pigmented terminal hairs.

38. Process for minimizing the rate of terminal hair loss in humans, comprising: topically applying a composition containing isolated omental lipids to human skin sites having areas genetically programmed for hair follicle growth.

39. Process of claims 36 or 38 wherein the topical application step further comprises: applying the composition at least once a day to the human skin sites having the areas genetically programmed for hair follicle growth.

40. Process of claims 36 or 38, comprising using the composition having isolated omental lipids in a range of from 5% to 20% by weight of the composition.

41. Process of claim 40 comprising topically using the composition having 5% to 20% by weight of isolated omental lipids of the total weight of the composition together with sodium hyaluronate.

42. Process of claim 40 further comprising topically applying the composition having isolated omental lipids incorporated into a lotion, a cream, a spray, a gel or a stick to the genetically programmed areas for hair follicle growth.

43. Process of claim 42 further comprising: topically applying the composition having isolated omental lipids and at least one member of the group consisting of sodium hyaluronate, an emulsifier, a hygroscopic agent, a preservative, a skin filler, a metal chelating agent and a mixture thereof to the human skin areas programmed for hair follicle growth.

44. Process of claim 42 further comprising: topically applying the composition having isolated omental lipids and at least one member of the group consisting of sorbitan stearate, a propylparaben, a polysorbate 60, a glyceryl stearate, a tocopheryl acetate, a vitamin A, a vitamin D, a dimethicone, a carbomer 940, water, a butylene glycol, a methylparaben, a trisodium EDTA, a phenoxyethanol, a triethanolamine, and a mixture thereof to the human skin sites programmed for hair follicle growth.

45. Composition comprising isolated lipids for application to balding human skin to reduce terminal hair loss or promote new hair growth.

46. Composition of claim 45 wherein the composition promotes new hair growth on human skin sites having areas genetically programmed for hair follicle growth.

47. Composition of claim 46 wherein new hair growth comprises terminal hair.

48. Composition of claim 45 wherein the lipids are derived from plant or animal tissue.

49. Composition of claims 45 or 48 wherein the lipids are isolated by a technique of the group of a cryogenic technique, extraction with at least one organic solvent, supercritical gas extraction, detergent extraction and heat extraction.

50. Composition of claim 45 further comprising sodium hyaluronate.

51. Composition of claim 45 further comprising at least one of the group of a silicone, an emulsifier, an antioxidant, vitamin A, vitamin D, vitamin E, a preservative, a hygroscopic agent, a metal chelating agent, a skin filler and a mixture thereof.

52. Composition of claim 45 wherein .the composition reduces terminal hair loss in humans having common pattern baldness.

53. Composition of claim 52 wherein the composition promotes new hair growth in humans having common pattern baldness.

54. Composition of claim 53 wherein the composition reduces terminal hair loss or promotes new hair growth in male human subjects having common pattern baldness.

55. Composition of claim 45 wherein the lipid material is incorporated into a lotion, cream, gel, spray, or stick.

56. Composition of claim 55 wherein the lotion, cream ointment, gel, spray or stick consists of a mixture comprising: materials selected from the group consisting of lipid material, sodium hyaluronate, sorbitan stearate, propylparaben, polysorbate 60, glyceryl stearate, tocopheryl acetate, vitamin A, vitamin D, dimethicone, carbomer 940, water, butylene glycol, methylparaben, trisodium EDTA, phenoxyethanol triethanolamine and a mixture thereof.

57. Process for promoting the rate of new hair growth in humans comprising: topically applying a composition containing isolated lipids to human skin sites having areas genetically programmed for hair follicle growth.

58. Process of claim 57 wherein the new hair growth comprises pigmented terminal hairs.

59. Process for reducing the rate of terminal hair loss in humans, comprising: topically applying a composition containing lipids to human skin sites having areas genetically programmed for hair follicle growth.

60. Process of claims 57 or 59 wherein the topical application step further comprises: applying the composition at least once a day to the human skin sites having, the sites genetically programmed for hair follicle growth.

61. Process of claims 57 or 59, comprising using the composition having isolated lipids in a range of form 5% to 20% by weight of the composition.

62. Process of claim 61 comprising using the composition having 5% - 20% by weight of isolated lipids together with sodium hyaluronate.

63. Process of claim 61 further comprising topically applying the composition having isolated lipids incorporated into a lotion, a cream, a spray, a gel or a stick to the sites genetically programmed for hair follicle growth.

64. Process of claim 63, comprising: topically applying the composition having isolated lipids and at least one member of the group consisting of sodium hyaluronate, an emulsifier, a hygroscopic agent, a preservative, a skin filler, a metal chelating agent and a mixture thereof to the human skin sites programmed for hair follicle growth.

65. Process of claim 63 comprising: topically applying the composition having isolated lipids and at least one member of the group consisting of sorbitan stearate, a propylparaben, a polysorbate 60, a glyceryl stearate, a tocopheryl acetate, a vitamin A, a vitamin D, a dimethicone, a carbomer 940, water, a butylene glycol, a methylparaben, a trisodium EDTA, a phenoxyethanol, a triethanolamine, and a mixture thereof to the human skin sites programmed for hair follicle growth.