Patent Application
20250177470
This invention relates to the field of nutritional supplements, and more particularly, to a composition comprising a lipidosterolic extract of Serenoa repens used to promote urinary and prostate function in men with enlarged prostates, or reduce androgenic hair loss in a patient, and a method for extracting the lipid components and enhancing the free fatty acids from the Serenoa repens berries for prostate, skin, and hair health.
Saw palmetto (Serenoa repens) is a shrubby palm that grows in the southeastern United States and is prolific on pine flatwoods in peninsular Florida and southern Georgia. The saw palmetto grows a seed stalk (spadix) in late spring, which flowers and then develops a hard green fruit about the size of a small olive. The fruit ripens in the summer, turning shades of yellow, orange and black. The ripe fruit is harvested by hand during the late summer and early fall.
Saw palmetto fruit (berries) have been ingested by Native Americans for many centuries. Early in the 20th century, naturopathic physicians in the U.S. began prescribing a tea made from saw palmetto berries as a mild diuretic and for genitourinary problems. It became a popular male tonic, but eventually the therapeutic value of the tea (a water extract) came into question, and in the early 1940's it lost both its popularity and its inclusion in the National Formulary. Research in Europe in the 1960's led to the identification of the beneficial aspects of the lipid portion of the berry and its medically active components, including free fatty acids and sterols.
Saw palmetto was listed in the U.S. Pharmacopeia from 1900 to 1916, and the National Formulary from 1925 to 1942. At various times from 1938 to 1990, Food and Drug Acts and Amendments were passed to address the problems of drug safety and effectiveness. Beginning in 1972, the FDA began evaluating over-the-counter drugs, including herbal preparations. In the U.S., however, it is permissible to sell herbal products only if no claims or statements regarding their value in the prevention or treatment of disease are made.
Saw palmetto berries are a common source for many modern nutraceutical formulations. When the saw palmetto berries are initially harvested, they are usually placed into standard citrus boxes, which are then loaded onto flatbed trailers, and transported by truck to drying facilities. The ripe berries contain approximately 66% water and are dried for several days at about 130-140° F., and after drying, may be stored for several years without deteriorating.
Dried berries are generally bagged and shipped to processing facilities where they are ground into a powder. A therapeutically effective dose can be prepared from a lipidosterolic extract of the dried berries. In Europe, ethanol or hexane solvents are commonly used to isolate the lipidosterolic extract from the berries, leaving a residue of the harsh organic chemicals remaining in the extract. These lipidosterolic extracts, such as the hexanic extracts, include fatty acids as various esters and free fatty acids, sterols and long-chain (fatty) alcohols. The fatty acids include caproic, caprylic, capric, lauric, myristic, palmitic, palmitoleic, stearic, oleic, linoleic and linolenic acid. Sterols include beta-sitosterol, campesterol, stigmasterol and stigmastanol. Long-chain alcohols include octacosanol, hexacosanol, triacontanol and tetracosanol. Usually, this extract is formulated into a convenient oral dosage form within a gelatin capsule.
Other techniques include the supercritical CO2 extraction of saw palmetto conducted at about 45° C. at about 220 bar to produce an extract SABALSELECT® that may be used without further purification, and includes fatty acids, alcohols and sterols. These saw palmetto extracts vary widely in their final composition with free fatty acids ranging in concentration from about 40% to below about 80%, methyl and ethyl esters ranging from about 1.5% to 16.7%, and glycerides ranging from about 6.8% to about 52.2%.
The fact that the concentration of various fatty acids, esters, and glycerides can vary in a saw palmetto extract makes it challenging to determine the effectiveness of any single saw palmetto extract, thus requiring a biological assay of each manufactured lot produced as a saw palmetto extract. To maintain continuous biological assays of each manufactured lot of saw palmetto extract to determine the impact on efficacy from changes in components, such as individual free fatty acids and ratios of free fatty acids to total fatty acids is complex, inefficient and expensive. Better control over pre-extraction growth, harvesting and processing together with controlled extraction would be advantageous that can be used to obtain a consistent, effective standardized lipid profile.
Lipidosterolic extracts of Serena repens (LSESr), may be sold alone, generally in 320 mg softgels, or in a variety of herbal formulations to support urinary and prostate health in men with prostate enlargement. In some instances, saw palmetto extracts have also been used for reducing androgenic hair loss in a patient. Certain saw palmetto extracts, such as Permixon®, are regulated as a prescription drug in France and Germany, and are used by many patients for treatment of benign prostatic hyperplasia (BPH), or enlarged prostate, but has drawbacks as a hexane extract containing residual solvent.
In the United States, however, lipidosterolic extracts of Serenoa repens dried fruit are considered dietary supplements which may also provide some health benefits, including benefits associated with promoting urinary and prostate function, improve urinary flow and control, or address hair loss in a patient, including Androgenic Alopecia (AGA), or to promote healthy hair growth. The currently available saw palmetto products, such as Permixon®, are usually lipid extracts removed as the whole lipid composition of the saw palmetto berry. Traditionally, extraction processes have not allowed for the formulation of a saw palmetto extract that include a desired blend of the various lipid fractions, and therefore, it was challenging to custom formulate a LSESr composition to meet customer specifications and have optimal therapeutic value. Although some CO2 extraction processes have yielded beneficial saw palmetto extracts, further control to enhance the percentages and ratios of compositional compounds, including different free fatty acids with a reduction in esters and glycerides, and an extended shelf life and low peroxide value without added antioxidants and stabilizers, would be beneficial for specific applications, such as prostate function, hair growth and health, skin care, such as acne remedies, and other applications.
A composition may be formulated in a therapeutic amount to maintain and promote urinary and prostate function in a human. The composition may comprise a shelf stable, supercritical CO2 fluid extracted, enhanced lipidosterolic extract of Serenoa repens (LSESr). The enhanced LSESr may comprise a ratio of free fatty acids to total fatty acids that is greater than about 83.0%, and an enrichment of lauric, myristic, oleic and linoleic acids as free fatty acids to total free fatty acids that is about 81.0% or greater.
The enhanced LSESr may have enrichment of lauric, myristic, oleic and linoleic acids as free fatty acids to total fatty acids that is greater than about 70.0%. The enhanced LSESr may have an enrichment of lauric, myristic, oleic and linoleic acids as free fatty acids that is greater than about 65.0%. The composition may include any one or more of zinc, vitamin D, biotin, collagen, rosemary oil and olive oil. The enhanced LSESr may have a balance of about 90.0% of total fatty acids to about 72.0% to about 75.0% of free fatty acids.
The enhanced LSESr may have an enrichment of lauric, myristic, oleic and linoleic free fatty acids to total free fatty acids of about 81.0% to about 84.0%. The enhanced LSESr may have a ratio of free fatty acids to total fatty acids from about 83.0% to greater than about 90%. The enhanced LSESr may have a ratio of free fatty acids to total fatty acids that is greater than about 908. The enhanced LSESr may have a peroxide value that is less than about 3 meg/kg and a shelf stability of at least about 4 years without added antioxidants and stabilizers. The composition may be formulated into an oral dosage form and in a capsule of about 70 mg to about 320 mg. The composition may include greater than about 0.2 percent w/w of saw palmetto sterols. The composition may include a dry excipient comprising at least one of silicon dioxide, calcium silicate, calcium phosphate, magnesium oxide, magnesium carbonate, calcium carbonate, rice fiber, and maltodextrin.
A method aspect to maintain and promote urinary and prostate function in a human includes administering a therapeutic amount of a composition to the human in need thereof. The composition may comprise a shelf stable, supercritical CO2 fluid extracted, enhanced lipidosterolic extract of Serenoa repens (LSESr). The enhanced LSESr may comprise a ratio of free fatty acids to total fatty acids that is greater than about 83.0%, and an enrichment of lauric, myristic, oleic and linoleic acids as free fatty acids to total free fatty acids that is about 81.0% or greater.
Other objects, features and advantages of the present invention will become apparent from the detailed description of the invention, which follows when considered in light of the accompanying drawings in which:
The present invention will now be described more fully hereinafter with reference to the accompanying drawings, in which preferred embodiments of the invention are shown. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
A method of processing saw palmetto berries to form a lipidosterolic extract as a dietary supplement composition by supercritical fluid extraction using CO2 under high pressure is illustrated in the high level flowchart of
The LSESr of the current invention is an enhanced LSESr that is improved over hexane solvent extracts such as commercially available Permixon® that contains residue of hexane and as it is environmentally friendly process. For purposes of description, the LSESr of the current invention will be described as an enhanced LSESr and has a favorable and advantageous 1) ratio of free fatty acids to total fatty acids; 2) higher enrichment of the free fatty acids of lauric, myristic, oleic, and linoleic as free fatty acids to total free fatty acids; and 3) specific free fatty acids tailored for specific end uses, such as more enhanced linoleic and linolenic free fatty acids such as above 3.7% of total free fatty acids for hair health, and such as in uses for prostate, hair health, and skin care with other free fatty acids. It has been found as explained further below that pre-extraction processing of the saw palmetto berry before the CO2 extraction has an important role in the final enhanced LSESr free fatty acid and fatty acid components that gives the best support for prostate, hair, and skin uses.
For example, hair happens to have numerous androgen receptors and the enhanced LSESr is especially useful for inhibiting the activity of 5α-Reductase-1 and 2 in certain individuals compared to the scalp of normal individuals, which has been seen to have a beneficial effect on hair health. The enhanced LSESr in an example, such as described below with reference to
For example, enhanced berry processing and specific extraction and separation pressure, temperatures and even times may be selected to form a final product as the enhanced LSESr of the current invention as described above, which also provides for a peroxide value that is less than about 3 meq/kg and a shelf stability of at least 4 years without added antioxidants and stabilizers. Generally, the enhanced LSESr is a shelf stable, supercritical CO2 fluid extracted, enhanced lipidosterolic extract of Serenoa repens (LSESr), the LSESr having a ratio of free fatty acids to total fatty acids that is greater than about 80.0% and an enrichment of lauric, myristic, oleic and linoleic acids as free fatty acids to total free fatty acids that is greater than about 82.0%. In an example, the enhanced LSESr may have a balance of about a preferred 90.0% of total fatty acids to about 72.0% free fatty acids, and in a specific example, about 88.9% to about 90.2% of total fatty acids to about 72.4% to about 72.6%. The enhanced LSESr may have an enrichment of lauric, myristic, oleic and linoleic free fatty acids to total free fatty acids of about 82.0% to about 84.0%. The enhanced LSESr may have a ratio of free fatty acids to total fatty acids of about 80.0% to about 82.08. These are non-limiting ranges and the values may range from about 2 to 3% above and below these ranges.
The enhanced LSESr may be used in a composition formulated in a therapeutic amount to maintain and promote hair health and growth in a human in need thereof. In this example the composition is a shelf-stable, supercritical CO2 fluid extracted, enhanced LSESr that has a ratio of free fatty acids to total fatty acids that is greater than about 80.0% and an enrichment of lauric, myristic, oleic and linoleic acids as free fatty acids to total free fatty acids that is greater than about 82.0%, and an enrichment of linoleic and linolenic acids together is greater than about 3.7%. The composition may be formulated as a topical lotion to be applied to the hair. The composition may include any one or more of zinc, vitamin D, biotin, collagen, rosemary oil and olive oil. The composition may include up to about 5.0% of Minoxidil.
The enhanced LSESr is preferably not contained on a carrier such as PEG (polyethylene glycol) to stabilize the free fatty acids, as compared to the commercially available Permixon®, which is on the PEG carrier. In addition to the four active free fatty acids listed above, a fifth as linolenic free fatty acid has been found to be advantageous to help with 5α-Reductase Type 1 as well as 5α-Reductase Type 2 as an inhibitor for hair treatment. The enhanced LSESr is produced using the inventive preprocessing, extraction and separation parameters to deliver the most active form of the free fatty acids for prostate and hair health. Use of preprocessing and pre-extraction processing, such as specific grinding and blending, gives better plating and densifies the powder. The front-end processing and aging of berries and attendant time sequencing of the saw palmetto powder after grinding allows for the exposure of the internal cell structure that helps the overall reaction and end product. These parameters include micromilling under cryogenic conditions and holding or ageing it.
Free fatty acids of the enhanced LSESr are better localized to the prostate, especially as a food source over glucose. The enhanced LSESr inhibits both the 5α-Reductase-1 and 2 enzymes that are especially beneficial for hair growth, and inhibits the 5α-Reductase-2 enzyme that is especially beneficial for prostate health. The free fatty acids of the enhanced LSESr localize to tissues with high high androgen receptor activity, like the prostate or the scalp, and can impact androgen receptors like 5α-Reductase 1 and 2. The preprocessing parameters as described below in greater detail help open up the berry matrix. The free fatty acids are better able to insert into the nuclear membrane with its attendant inhibitory effects on 5α-Reductase to prevent conversion of testosterone to DHT. Normal and routine extraction of saw palmetto berries produces usually about 70% free fatty acids or less, which is well below the percentage of free fatty acids in the enhanced LSESr of the current invention.
Androgens play an important role in prostate growth and development and in hair growth. Chief among these androgens are testosterone and dihydrotestosterone (DHT). The 5α-reductase (5αR) enzymes are responsible for converting testosterone into the more potent dihydrotestosterone (DHT). There are three different types, or isoforms, of the 5α-reductase (5αR) enzymes: 5αR-1, 5αR-2, and 5αR-3. These isoforms can be found in various tissues and in varying amounts throughout the body.
However, 5αR-2 in particular can become overexpressed in prostates of aging men leading to prostate enlargement. Meanwhile, both 5αR-1 and 5αR-2 can become overexpressed in the scalps of men or women who are genetically susceptible to androgenic hair loss. As will be explained in greater detail below with reference to
Both testosterone and DHT can bind to and activate androgen receptors, but DHT is more potent. Activation of androgen receptors leads to gene expression. Since DHT is more potent, it induces higher levels of gene expression when it activates androgen receptors, compared to the less potent testosterone. Overexpression of 5αR enzymes in the prostates of aging men or in the scalps of individuals susceptible to androgenic hair loss leads to the overproduction of DHT. More DHT means an even greater response in gene expression. That imbalance in gene expression can then lead to abnormal function in the tissue, like prostate enlargement or hair loss.
Certain fatty acids found in saw palmetto extract can inhibit the 5αR enzymes in the prostate and scalp as shown in
The combination of the free fatty acids plus bound fatty acids is the total fatty acid level.
Identification: Unlike animal oils or most vegetable oils which contain fatty acids mainly in the form of triglycerides, saw palmetto oil extract is rich in free fatty acids. One method of identifying authentic saw palmetto extracts from imitation or adulterated extracts is to test the free fatty acid levels as a percentage of the total fatty acid levels. High-quality saw palmetto extracts contain ≥70% free fatty acids.
Absorption: Free fatty acids are absorbed twice as well as triglycerides.
Disposition: Once absorbed into the body, free fatty acids tend to localize to the prostate and scalp. Why? Because cells with high expression of androgen receptors, like those in the prostate or scalp, also express fatty acid transporters on the cell surface. Those transporters preferentially pull free fatty acids into the cell instead of bound fatty acids.
Activity: Free fatty acids are the active ingredients in saw palmetto extracts. Certain free fatty acids are potent inhibitors of the 5αR enzymes, versus other forms of fatty acids which are inactive, as shown in
In particular, lauric, myristic, oleic, linoleic, and linolenic acids appear to be most effective at inhibiting 5αR enzymes in the prostate, while linoleic, linolenic, and oleic acids seem most effective at inhibiting 5αR enzymes in the scalp. Therefore, increasing the overall free fatty acid levels in an LSESr enhances the quality, the absorption, and the localization of fatty acids to the prostate and scalp. Increasing the levels of certain free fatty acids should enhance the potency of the LSESr in specific tissues.
An LSESr that is more effective at inhibiting 5αR enzymes in the prostate should be more effective in maintaining normal urinary and prostate function in aging men than a standard extract. An LSESr that is more effective at inhibiting 5αR enzymes in the scalp should be more effective in promoting healthy hair growth in individuals susceptible to androgenic hair loss than a standard extract.
The preprocessing and milling help obtain a desired particle size and surface area where about 10% is less than about 4 mesh and about 95% less than about 12 mesh powder and help open up the matrix to optimize the free fatty acids as in the enhanced LSESr. Some prior art CO2 extraction parameters operate at 100-350 bar, which has not been found adequate, especially since the process may be single step. Chemical transformation and alkyl hydrolysis has not been found beneficial since it may erode quality.
The enhanced LSESr of the current invention aids in balancing androgen signaling and controlling inflammation. The free fatty acid oleic acid has been found especially effective for 5α-Reductase-1, which is important for hair health, and not as much for 5α-Reductase-2, while linoleic free fatty acid has been found beneficial for both, which is beneficial for hair health treatment. Esterified fatty acids have been found ineffective. Linolenic acid has been found beneficial for both 5α-Reductase-1 and 5α-Reductase 2, and thus effective in hair health. The preprocessing of the saw palmetto berries into the saw palmetto powder helps open up the cell structure, and the aging, such as at least 15 days, and in an example, about 15 to 30 days of ageing, together with other factors, helps give the saw palmetto powder a beneficial moisture level, which aids during the CO2 extraction and separation form the enhanced LSESr. There are positive benefits such as the enhanced LSESr aiding in stem cell signaling, which helps influence hair growth and the hair growth cycle.
For hair health, the inventors have discovered that linoleic and linolenic free fatty acids are beneficial and more active than previously understood by those skilled in the art. It is known that DHT may inhibit WnT signaling, which among the signaling molecules, promotes the differentiation of hair follicle stem cells. The enhanced LSESr helps block 5α-Reductase activity thereby reducing release of DHT that would otherwise inhibit WnT signaling, which plays a key role in simulating hair follicle stem cell regeneration.
An example of producing a LSESr which may be processed for an enhanced LSESr is now described. As shown in the flowchart, dried saw palmetto berries are supplied (Block 102), fed into a feed hopper (Block 104), and metallic particles and other magnetic particles or objects may be magnetically separated from the berries (Block 106). A hammer mill may be used to crush and mill the dried saw palmetto (Block 108). Cooling may be maintained to prevent excessive heating during milling by the application of liquid nitrogen to the hammer mill as a non-limiting example (Block 110). CO2 extraction begins (Block 112) using carbon dioxide in an example (Block 114). Spent marc (Block 116) may include some organic residue and have some end use purposes, and thus, may be retained or discarded. CO2 extraction includes a first fractionation (Block 118) and forming a first extract (Block 120), followed by a second fractionation (Block 122), and forming a second extract (Block 124), and a third fractionation (Block 126), and forming a third extract (Block 128), with recycling of the carbon dioxide (Block 114).
The method as described generally includes drying the berries, grinding the dried berries, extracting the berries with the CO2 under relatively high pressure, and separating the extracted components from the CO2. When ripe berries are harvested, they may contain about 66% water and may be dried for several days at 130-140° F. In the resulting dry state, the berries may be stored for several years without further deterioration. Dried berries may be bagged and shipped to processing facilities where they are ground to a fineness where about 100% are less than about 4 mesh and about 95% less than about 12 mesh into a fine powder.
Supercritical CO2 fluid extraction may be accomplished in an extractor vessel by contacting the ground saw palmetto berries as a fine saw palmetto powder with a continuous flow of CO2 at an extraction pressure of at least about 300 bar, and at a temperature lower than about 85° C. to thereby extract saw palmetto components from the CO2. The process includes separating the extracted saw palmetto components from the CO2 in a series of separator vessels by collecting the carbon dioxide after extraction and decreasing the pressure in a decreasing, stepwise manner.
In an example, the extraction vessel may be loaded with the ground, dried saw palmetto berries as a saw palmetto powder, and the CO2 may be delivered into the vessel in a manner where the CO2 flows through the loaded product before being discharged from the vessel. As the high-pressure CO2 flows through the product, it behaves as a lipophilic solvent and extracts those saw palmetto lipidosterolic components that are soluble in the fluid. Different extraction pressures may be used, but it has been found that an extraction pressure of about 500-550 bar is advantageous. A first separation pressure may be about 250-300 bar, and may be about 160 bar to about 350 bar. Extraction may be conducted at a temperature from about 45° C. or 50° C. to about 90° C., and preferably under a substantially continuous flow of carbon dioxide. In an example, the first fractionation (Block 118 in
The method may include the second fractionation (Block 122 in
Additionally, in accordance with another embodiment, the method may include a third fractionation (Block 126 in
Plural separations may be conducted sequentially and in a substantially continuous flow. Each subsequent separation may have a lower predetermined separation pressure and operate as sequential separations referred to as cascading separations. Sequential separations separate the extracted saw palmetto compounds from the carbon dioxide into a plurality of fractions. Passing the substantially continuous flow of carbon dioxide after extraction through the different separations will separate the extract into several fractions.
In a preferred technique, each individual fraction of the resulting plurality of fractions may have a different fraction of extracted saw palmetto components from the other individual fractions. For example, the first fraction may include a major fraction of saw palmetto sterol and triacylglyceride compounds, the second fraction may include a fraction of saw palmetto components similar to a whole extract, and a third fraction may include a major fraction of saw palmetto unesterified, free fatty acids. This is contrasted where acylglyceride compounds include esters derived from glycerol and 1-3 fatty acids.
An advantageous benefit of the method as described is the ability to blend the resulting extraction fractions at predetermined amounts to prepare a dietary supplement composition that includes desired proportions of different saw palmetto components, including sterols, unesterified free fatty acids and fatty alcohols. This method allows not only the preparation of custom blends to meet specific consumer demands, but also the preparation of a dietary supplement composition that has been standardized for the final end use of the product.
The resulting dietary supplement composition may be formulated for oral use and nutritional supplementation. The composition may include a pharmaceutically acceptable carrier and may be orally administered as a gel capsule containing a unit dose. In an example, the daily dose may contain about 160 mg b.i.d. to about 320 mg, or at least about 200 mg of saw palmetto lipids as an enhanced lipidosterolic extract of Serenoa repens, (LSERs), and in another example, the unit dose may contain about 320 mg of saw palmetto lipids as enhanced effectiveness from LSERs for a single daily unit consumption.
A method for use may include ingesting the dietary supplement composition having at least about 85% saw palmetto fatty acids as the enhanced LSESr, and preferably greater than about 90% of these fatty acids present as free, unesterified fatty acids. The method for use may include ingesting the dietary supplement composition that has essentially no solvent residue. Ingesting may be accomplished by swallowing a capsule unit dose that includes a pharmaceutically acceptable carrier. A suggested daily dose may be about 320 mg of saw palmetto lipids as a LSESr, or about minimum 200 mg per dose of enhanced LSESr, preferably having a relatively high amount of unesterified fatty acids greater than about 72%.
An example of the production parameters and resulting extraction components from the extraction and isolation of the saw palmetto lipids based upon the process described relative to the flowchart in
During the supercritical carbon dioxide fluid extraction, it may be possible to use entrainers as cosolvents, although this is not necessary to obtain the desired LSESr. It is also possible to modify the phase behavior of supercritical solvents and enhance solubility of the components in the carbon dioxide-rich phase. A cosolvent may alter the polarity, viscosity and density of any gas phases. It may be possible to use cosolvents such as alcohols and hydrocarbons (straight-chained and branched) that may have a higher molecular weight. It may be possible to use ethoxylated compounds.
The enhanced lipidosterolic extract of Serenoa repens (LSESr) as manufactured and described may be formulated for prostate health or hair health, including addressing androgenic hair loss. The enhanced LSESr has been shown to inhibit 5α-Reductase 1 and 2. The enhanced LSESr may specifically target prostate tissue, reduce urinary frequency, and support, maintain and promote healthy urinary and prostate function. The enhanced LSESr may support optimal urinary flow and control in men and inhibit 5α-reductase without sexual side effects.
This enhanced lipidosterolic extract of Serenoa repens may be formulated for hair health treatment and regrowth because the enhanced LSESr has been found to target 5α-Reductase 1 and 5α-Reductase 2, which are found in hair follicles and affect hair regrowth and thickness.
The enhanced LSESr has a high percentage of free fatty acids not commonly found in nature. The enhanced LSESr has markedly different characteristics with a marked change in function and activity to treat symptoms of prostate disorder and promote urinary and prostate function, or for hair loss in a patient to promote prostate health and hair regrowth. The different free fatty acids as fractions alone or in combination provides a marked change in function and activity compared to what is normally found in nature. These benefits are accomplished via the plurality of separations and resulting manufacture of several fractions.
The enhanced lipidosterolic extract of Serenoa repens (LSESr) as manufactured and described may be encapsulated in a gelatin, carrageenan or starch based soft gel for oral consumption. The enhanced LSESr may be combined with dry excipients including, but not limited to one or more of silicon dioxide, calcium silicate, calcium phosphate, magnesium oxide, magnesium carbonate, calcium carbonate, rice fiber, and maltodextrin to create a free-flowing powder form of the extract. The enhanced LSESr may also be combined with excipients and spray dried into a free-flowing powder. The enhanced LSESr may also be combined with water and emulsified using excipients to create a stable emulsion, which may be dried such as by spray drying, belt drying, vacuum drying, or freeze drying to create a powder form.
The different carriers and formulations may also impart markedly different characteristics for the enhanced LSESr to stabilize the composition and impart functions that are significantly more in function than found in nature, and help in bioavailability of the composition that includes the enhanced LSESr when orally digested. It is possible to use a carrier for contacting the skin to help reduce acne.
Other additives may be included with the enhanced LSESr and may include pumpkin seed, Vitamin D, zinc, biotin, collagen, olive oil and rosemary oil, alone or in different combinations. The enhanced LSESr has been found to be stable and not require added antioxidants and has extended shelf life. Other additives may be included, such as Valensa's O2B® Peroxidation Blocker stabilization technology and other components, including astaxanthin, phenolic additives, and natural and synthetic tocopherols and tocotrienols, carnosic acid or carnosol and/or astaxanthin, but even without these additives, the enhanced LSESr has an extended shelf life as noted before.
Other additives with the enhanced LSESr may include a mixture of selected lipophilic and hydrophilic components. Lipophilic additives may be used either alone or in combination with at least one of: a) phenolic additives including at least one of sage, oregano, and rosemary; b) tocopherol(s); c) tocotrienol(s); d) carotenoids including at least one of astaxanthin, lutein, and zeaxanthin; e) ascorbyl acetate; f) ascorbyl palmitate; g) Butylated hydroxytoluene (BHT); h) Butylated hydroxyanisole (BHA); or i) Tertiary Butyl hydroquinone (TBHQ). A hydrophilic additive may include a sequestrant and may include hydrophilic phenolic additives including at least one of grape seed extract, tea extracts, ascorbic acid, citric acid, tartaric acid, and malic acid.
The process for manufacturing the enhanced LSESr as described above, such as with reference to the flowchart of
An example of the enhanced LSESr is shown in the table of
The major contributors of active fatty acids unique to the enhanced LSESr are largely free fatty acids. The free fatty acids in the enhanced LSESr are uniquely intensified compared to other forms of fatty acids found in other sources. Vegetable oils contain fatty acids primarily in the triglyceride form, whereas LSESr contain primarily free fatty acids. In addition, LSESr contain high levels of lauric acid. Most vegetable oils, with the exception of coconut oil, typically do not contain lauric acid.
It is known that free fatty acids, such as derived from fish oil, are better absorbed and more bioavailable than corresponding triacylglycerides or ethyl esters that may be found in fish and similar oils. For example, absorption may occur at greater than 95% for the free fatty acids, versus 57% to 68% for the triacylglycerides, or versus 20% to 21% for the ethyl esters, respectively. There are some saw palmetto extracts that have high levels of free fatty acids, while other saw palmetto extracts have lower levels of free fatty acids. For example, depending on the type of extraction technique applied to saw palmetto berries or ground saw palmetto, the levels of free fatty acids in saw palmetto extracts may vary from 40% to 80%, but typically never beyond 80% even with supercritical CO2 extraction. Saw palmetto extracts can be expensive to produce, and for that reason, there are known vegetable oils that are commonly used to adulterate or imitate saw palmetto extracts, but these oils contain high levels of triglycerides. Those saw palmetto extracts having higher free fatty acid concentrations are better absorbed and made more bioavailable than saw palmetto extracts having lower levels of free fatty acids and vegetable oil comparators. Once absorbed in the body, some free fatty acids specifically target the prostate, or the scalp to aid in hair health and hair growth.
There are some factors making free fatty acids undesirable in vegetable oils, even though they are made more bioavailable when ingested, for example. Free fatty acids are highly reactive and may create stability issues, and vegetable oils may degrade quickly. For example, heat and frying may degrade vegetable oils, and thus, quality is monitored by testing peroxide value as a measure of oxidative degradation.
This issue of oxidative degradation is not as challenging with saw palmetto extracts because the oil extracted from the saw palmetto is acidic, unlike vegetable oils, which has a more neutral pH. This benefit has been realized by the inventors with the enhanced LSESr of the invention. For example, the enhanced LSESr has a four-year shelf life that is a technical achievement for an oil that is high in free fatty acids, and especially with the higher ratio of free fatty acids to total fatty acids. This higher ratio of free fatty acids to total fatty acids and especially the contribution of four free fatty acids of lauric, myristic, oleic and linoleic was accomplished by the unique preprocessing of saw palmetto berries in combination with the unique supercritical CO2 processing parameters. It is feasible to achieve high free fatty acid oil concentrations with an acidic pH, and this benefit of higher concentration is aided because the saw palmetto berries act as a natural oil protection, while excluding oxygen, without having to artificially stabilize the free fatty acids. For example, in an example of the enhanced LSESr, the peroxide value (PV) specification is below 5.0 meq/kg, and is below 1.0, so oxidative damage is controlled as well.
Based on a lipid chemistry perspective, the higher the free fatty acids are driven, the more likely there will be stability issues encountered. However, in accordance with a non-limiting example of the invention, the preprocessing of saw palmetto berries as described with the improved supercritical CO2 extraction and enhanced processing parameters, the stability issue because a more minor issue because the supercritical CO2 extraction process is a closed system in the absence of oxygen and used without organic solvents, such as hexane. This may be distinguished with a known commercial extraction with the organic solvent hexane as used to manufacture the commercially available European extract known as Permixon® with its attendant technical drawbacks as explained below.
Enhanced processing parameters for supercritical carbon dioxide extraction is employed when manufacturing the enhanced LSESr of the current invention rather than hexane, and this permits the process to achieve a stable extract which is “naturally/non-organic solvent” stable. Together with unique preprocessing of the saw palmetto berries, the enhanced LSESr is produced. Thus, any composition that includes the enhanced LSESr achieves a balanced fatty acid profile that is rich in free fatty acids, room temperature stable and stable for years.
Referring now to the tables shown in
The conclusions for the biological assay are shown in
If the concentration of free fatty acids is too high, it may be more difficult to obtain enough enrichment in the four important fatty acids that drive biological activity, i.e., the lauric, myristic, oleic, and linoleic free fatty acids. For that reason, one goal was to achieve the range of about 82.0% to about 84.0% for these four bioactive free fatty acids. This can range from about 82.0% to as high as 86.0%, but preferably about 82.0% to about 84.0%. The better optimization comes from supercritical CO2 extraction control and balance, such as described with reference to the process shown and described relative for the process shown in
Experiment 1 as U4868 was a lower pressure and temperature extraction process such as typically used for extraction of hops products, e.g., around 150 bar and 40° C. The Experiment 1 low pressure process had quality issues by not removing all components, such as the mono and diglycerides and unsaponifiable matter. Although it may develop a ratio of free fatty acids to total fatty acids greater than the commercially available Permixon® and even some fatty acid ratios similar to the enhanced LSESr of Experiments 5 and 6, the lower pressure process produced an extract product differing from the invention in Experiments 5 and 6. The greater than 80% ratio of free fatty acids to total fatty acids and higher percentage of the four active free fatty acids was achieved for the enhanced LSESr shown by Experiments 5 and 6 as Lot Nos. 211022 and 211105, and had excellent shelf stability and as much as 4 years stability without added antioxidants and stabilizers. This was also achieved by the enhanced berry preprocessing with aging of the cryogenically milled saw palmetto berries and the CO2 extraction parameters as later described.
Experiment 3 had similar CO2 extraction processing parameters with slight differences, but no enhanced preprocessing of berries with specific cryogenic milling and aging of saw palmetto powder as with Experiments 5 and 6. Thus from those test results, it was evident that biological activity for the LSESr of Experiments 5 and 6 was not driven by simple factors of total fatty acids, free fatty acids, or the level of the important fatty acids as free fatty acids. The better profile of the enhanced LSESr was driven by the more desirable and inventive pre-extraction handling parameters described below and the inventive supercritical CO2 extraction and separation parameters on the ripe berries, such as described relative to the flowchart of
Comparing the percentages across the different lots, the total fatty acids is less important than the ratio of total fatty acids to free fatty acids. What is also important is the optimization of the contribution of the four bioactive fatty acids (lauric, myristic, oleic, and linoleic) as a percentage of total free fatty acids. These beneficial values in the enhanced LSESr are driven by the inventive preprocessing parameters for the saw palmetto berry and supercritical CO2 extraction parameters. A fifth bioactive free fatty acid, linolenic, also contributes especially for hair health and operates to help reduce inflammation and aid signaling. It is possible to achieve a similar percentage of free fatty acids to total fatty acids from proportionally lower total fatty acids or higher free fatty acids. For example, Experiment 1 as the low pressure process (U4868) delivered more of these total free fatty acids at 87.0% and the contribution of free fatty acids to total fatty acids at a higher 94%, but less than 82.0% for the active four free fatty acids as in Experiments 5 and 6.
The aging, milling, and particle size deliver the optimum ratio of 4 (four) critical free fatty acids to total fatty acids via the supercritical CO2 extraction. The invention and optimized aging/moisture, particle size and extraction parameters drive the value of the four active free fatty acids to total free fatty acids, not dependent on achieving highest total fatty acids or free fatty acids. The preconditioning of the berries allows the extraction process to achieve the inherently beneficial profile.
The 5α-Reductase Type 2 50% (IC50) inhibition data are evident in the graphs and bar charts of
In Inhibition Experiment 1 (
Inhibition Experiment 2 shown in
Lipid extracts manufactured using conventional techniques such as hexane extraction (Permixon®) or ethanol extraction (SABALSELECT®) from ripe saw palmetto berries typically contain over about ca. 70% free fatty acids. The experiments and studies show that ripe saw palmetto berries that have been preprocessed using techniques of milling and grinding as developed by the inventors and coupled with selected CO2 extraction and separation pressures and temperatures as developed by the inventors may be employed to optimize the free fatty acids, which has been found to be a key to effectiveness, such as accomplished with the enhanced LSESr of Experiments 5 and 6 as Lot Nos. 211022 and 211105. Both the total and free fatty acids are important, but the higher level of free fatty acids and especially the four bioactive free fatty acids of lauric, myristic, oleic and linoleic are more biologically more relevant. Experiments have also shown that only certain types of fatty acids inhibit 5α-Reductase isoenzymes Type 1 and Type 2, such as the four primary more bioactive free fatty acids of lauric, myristic, oleic, and linoleic acids, and with a fifth as linolenic free fatty acid in combination with linoleic acid, and each of the free fatty acids having a different effect whether it is 5α-Reductase Type 1 or Type 2 enzyme.
Linolenic acid has been found advantageous for hair health in combination with linoleic acid as free fatty acids and inhibiting 5α-Reductase Type 1 enzyme and as an aid in signaling. These free fatty acids are believed to act synergistically in inhibiting 5α-Reductase both Type 1 and Type 2. The overall lipid profile of the enhanced LSESr confirms authenticity. By establishing pre-extraction processing parameters of ripe berries, together with the production parameters for supercritical CO2 extraction, it is possible to ensure that the desired free fatty acid concentrations and ratios, especially with the four bioactive free fatty acids, are met to form the enhanced LSESr. Thus, a quality product may be produced such as the enhanced LSESr. Biological assays will not be required for every production lot since the repeated use of the preprocessing parameters of the saw palmetto berries and the specific supercritical CO2 extraction parameters will repeatedly produce the enhanced LSESr having a ratio of free fatty acids to total fatty acids that is greater than about 80.0% and an enrichment of lauric, myristic, oleic and linoleic free fatty acids that is greater than about 82.0%.
In an example, this enhanced LSESr may have a balance of about 90.0% of total fatty acids to about 72.0% of free fatty acids, and an enrichment of the four active free fatty acids of about 82.0% to about 84.08. It has been found the ratio of free fatty acids to total fatty acids of about 80.0% to about 82.0% is effective with about 82.0% to about 84.0% of the four bioactive free fatty acids.
The fatty acids in the LSESr target the prostate. The 5α-Reductase enzymes are found in several different tissues in the body. However, the free fatty acids in the enhanced LSESr target the prostate cells, and unlike most cells in the body, the prostate cells preferentially uptake these free fatty acids instead of glucose. The androgen receptors up regulate fatty acid transporters on the surface of prostate cells.
It has been found that there is a difference in function between Finasteride as a 5α-Reductase inhibitor and the enhanced LSESr bioactive free fatty acids. Finasteride permanently locks the enzyme closed, and competitively blocks the binding sites. The enhanced LSESr bioactive free fatty acids, on the other hand, temporarily change the make-up of the nuclear membrane, which disrupts the stability of the enzyme, and non-competitively blocks the binding sites. This is one reason why the type, quantity and ratio of the free fatty acids relative to fatty acids can be important and slight variations, such as found with the enhanced LSESr of the invention as compared to other CO2 extracted products or Permixon® or other hexane extracted saw palmetto extracts can have such a positive difference in function.
By specifically targeting prostate cells, the enhanced LSESr has no effect on circulating DHT (Dihydrotestosterone) levels and has low side effects, including low sexual side effects. The free fatty acids in the enhanced LSESr change the lipid composition of the nuclear membrane, which disrupts 5α-Reductase activity. The impact on the 5α-Reductase enzymes is reversible and non-competitive, while the biological function of the 5α-Reductase enzyme is conserved. These effects are not seen with vegetable oils or saw palmetto imitators because these oils have the wrong forms of fatty acids and free fatty acids, the wrong ratios, and the wrong lipid profile. Powdered saw palmetto products are very common, but they do not have enough lipid content, and often have an inferior level and type of fatty acids.
The free fatty acids are inserted into the prostate cell nuclear membrane and impact functionality of that cell nuclear membrane, disrupt 5α-Reductase enzyme activity, lead to no further or limited enlargement of the prostate, lead to less inflammation with a resultant improvement in urinary flow and control, and no delayed progression of the condition. Small changes in amounts, ratios and specific free fatty acids make large differences as the inventors have determined, such as with the four bioactive free fatty acids, enabling peak results for the enhanced LSESr. The free fatty acids of the enhanced LSESr operate as active ingredients for clinically proven anti-inflammatory and anti-proliferative effects, which are likely due to reduced inflammatory enzyme activity such as 5-lipoxgenase, COX-2, Phospholipase A2, and similar enzymes via disruption of the cell nuclear membrane. The inhibition of the NF-KB activation reduces inflammatory gene expression.
These effects and functions can be explained when comparing a balanced immune system and an unbalanced immune system. For example, a balanced immune system responds to triggers such as an infection or injury and activates pro-inflammatory enzymes and pro-inflammatory mediators to deal with the threat. A balanced immune system also resolves inflammation after the threat has been neutralized, which promotes healing of tissues by increasing production of growth factors. An unbalanced immune system, on the other hand, does not resolve inflammation, even after the threat is no longer present. An unbalanced immune system may have an uncontrolled activation of pro-inflammatory mediators, which leads to tissue damage and dysregulates production of growth factors, resulting in abnormal cell proliferation and tissue remodeling. Hypoxemia may occur from rampant cell growth that leads to angiogenesis and production of more growth factors.
The enhanced LSESr of the current invention disrupts the activity of pro-inflammatory enzymes and the expression of inflammatory genes. Similar to 5α-Reductase, the enhanced LSESr changes the composition of the cell nuclear membrane, which disrupts the activity of pro-inflammatory enzymes. By blocking activation of NF-KB, the enhanced LSESr reduces the expression of numerous inflammatory mediators such as pro-inflammatory enzymes, adhesion molecules, cytokines, chemokines, growth factors, receptors, and transcription factors.
It is understood that the fatty acid transporters on prostate cells preferentially take up the free fatty acids instead of glucose, unlike most cells in the body. Thus, the free fatty acids of the enhanced LSESr inhibit the 5α-Reductase and are anti-inflammatory. The enhanced LSESr of the current invention is rich in free fatty acids and their specific combination and ratio. An example is the enhanced LSESr of Experiments 5 and 6 (
Reference is made again to
Inhibition Experiment 2 (
It should be understood that the enhanced LSESr is also beneficial for hair treatment of adrogenic hair loss and reduces inflammation and aids in stem cell signaling. Hair growth and loss is a natural occurrence with the growth phase referred to as Anagen where the follicles are anchored at the base. This stage occurs for about 2-6 years duration and about 90% to about 95% of the follicles are at this stage of the hair growth cycle. The transition phase (Catagen) is about 2-3 weeks and less than 1% of hair follicles are at that stage. In this Catagen stage, the hair follicles slowly detach from nourishing blood supply and stop growing. The resting phase known as Telogen is about 2-3 months and about 5% to 10% of follicles are in this stage, where there is no nourishment and the hair dies and falls out. The early Anagen phase follows where the hair follicle re-establishes and hair regrowth begins. The duration of the growth phase imparts hair length and can be 2-6 years in duration. Blood flow provides nourishment that enables growth.
When the hair growth cycle is impacted, the result is hair thinning and resultant shorter and finer hair follicles over time, such as occurs with miniaturization and alopecia. Thus, any hair loss remedy must balance the cycle of growth, loss, and the resting phase before growth begins again. Humans are typically born with all hair follicles, e.g., about 100,000 on the scalp. More hair follicles are not produced, but hair growth and hair health can be supported.
Androgens are considered the main regulators of the hair follicle. Normal hair growth cycle takes place in the presence of androgens. However, androgens may inhibit hair follicle growth in genetically susceptible individuals, leading to small, thin hair and hair loss, also known as Androgenic Alopecia (AGA). Usually AGA is a hereditary thinning of the hair induced by androgens and may begin between the ages of 12 and 40 in both sexes, but it is less severe in women due to the lower levels of androgens.
There is an interplay between androgens and the androgen receptor and the hair follicle. Androgens from the blood, such as circulating testosterone, may enter the hair follicle via the dermal papilla's blood supply and may be converted by 5α-Reductase to the more potent androgen DHT, which may bind to the androgen receptors in the dermal papilla cells, causing changes in their production of regulatory paracrine factors. This directly alters the activity of dermal papilla cells. Keratinocytes and melanocytes may also be affected. The complex then binds to elements in the DNA, altering the expression of specific androgen-dependent genes.
If both testosterone and DHT are present in similar quantities, the androgen receptor will bind 5α-dihydrotestosterone (DHT) rather than testosterone. Once the 5α-DHT complex is formed, the receptor complex undergoes conformational change. Along with co-activating proteins, the 5α-DHT complex binds to elements in the DNA, altering expression of specific androgen-dependent genes. The effect is the hormone-receptor complex-activated genes change large terminal follicles to miniaturized follicles. Each successive hair cycle undergoes a shorter growth phase and the follicles become smaller, producing shorter, finer hair that poorly covers the scalp.
Signaling is indicative that the hair follicle stem cells move from the resting phase and start the growth phase. DHT interferes with the signaling that begins the hair growth cycle and can activate expression of an important gene (DKK-1), a major hair loss factor secreted from dermal papilla (DP) cells that may induce and accelerate AGA. DHT up-regulates Catagen inducers, and the dermal papilla cells produce DKK-1 in response to DHT in the AGA scalp.
DKK-1 is a Catagen inducer and suppresses dermal papilla cells and keratinocyte proliferation and slows the differentiation of hair follicle stem cells. DHT also up-regulates the secretion of IL-6 and T6F-β1 by dermal papilla cells. This suppresses keratinocyte proliferation and inhibits hair shaft elongation. Scalp DKK-1 is much higher in individuals with AGA than evident with others serving as controls. Men have more 5α-Reductase enzymes and androgen receptors in the scalp than women, which leads to higher levels of DHT-AR complex and higher DKK-1 levels in scalps. For that reason, androgenic hair loss in men is more severe than for women.
The 5α-Reductase enzymes Type 1 and Type 2 are located within hair follicles, where they influence hair growth and hair miniaturization. The 5α-Reductase Type 1 enzyme is almost everywhere, while the 5α-Reductase Type 2 is very localized. 5α-Reductase 1 and 2 are present in higher levels in women with female pattern hair loss and the 5α-Reductase inhibitor therapy has been shown to be effective in men with androgenic hair loss. The hair structure includes different components. The inner root sheath anchors and protects the hair shaft and the outer root sheath surrounds the entire hair follicle and is a source of stem cells. The matrix cells generate hair and the papilla allow for vasculature for blood supply. The more localized 5α-Reductase Type 2 enzyme is not typically located in the outer root sheath and papilla. For this reason, the enhanced LSESr may affect 5α-Reductase Type 1 enzyme, and having enhanced linolenic and linoleic free fatty acids, may be better suited for hair health.
Saw palmetto extract of about 320 milligrams per day versus finasteride of 1 milligram per day had been studied in 100 men with mild to moderate androgenic alopecia to assess changing hair density over 24 months. Both treatments increased hair growth but finasteride was found more effective. The saw palmetto extract stabilized hair loss in 52% of men, and 38% of men treated with the saw palmetto extract had increased hair growth, and 68% of men treated with finasteride noted an improvement.
However, as is well known, finasteride comes with side effects not seen with saw palmetto extracts. Inhibiting the 5α-Reductase Type 1 enzyme helps limit the production of DHT and supports hair health. The graphs in
Reference is now made to
The hair follicle includes lipids and especially fatty acids and free fatty acids. The highest concentration of lipids in human hair is about 4.3 mg/g of free fatty acids. For example, the medulla of the hair follicle includes squalene, triglycerides, cholesterol, wax esters, and free fatty acids such as oleic acid and palmitic acid. The cortex of the hair follicle includes integral fatty acids such as linoleic acid and alpha-linoleic acid and non-covalently attached fatty acids, triglycerides, cholesterol, wax esters, and squalene. The interface between the cortex and cuticle includes a lipid containing N-acetylglucosamine. The entire cuticle contains ceramides and cholesterol. An epicuticle includes 18-methyl eicosanoic acid (18-MEA) and an exocuticle includes free fatty acids and covalently-attached fatty acids. An endocuticle includes free fatty acids and covalently-attached fatty acids. Free fatty acids are common lipids in hair, mainly in the hair matrix cells.
Recent studies show that linoleic acid promotes hair growth in normal mice, and promotes the Anagen phase in normal shaved mice by up-regulating hair growth related proteins. After about 20 days, linoleic acid operated better than controls and equivalent to Minoxidil with about 100% hair growth.
Referring now to
It is known that oral free fatty acids may be localized to the skin and hair follicles in addition to the prostate where the five alpha-reductase enzymes are located. The human hair contains a high percentage of fatty acids, mainly in the free fatty acid form. Thus, it is possible that some of the linolenic acid may have instead become incorporated into the fur of the guinea pigs. The enhanced benefit of the free fatty acid linolenic acid for hair health and hair growth is evident.
The enhanced LSESr of the current invention may also be used to reduce acne since there is localization of 5α-Reductase enzyme in inflammatory acne lesions and inhibition of 5α-Reductase has been shown to improve acne in men with androgenic hair loss. 5α-Reductase Type 1 enzymes are the predominan isotype in acne lesions and are localized in the sebaceous glands. 5α-Reductase Type 2 enzymes are the minor isotype in acne lesions and are localized to sebaceous duct and endothelial cells.
The enhanced LSESr of the current invention inhibits 5α-Reductase 1 and 2 enzymes that play an important role in androgenic hair thinning and eventually hair loss. The enhanced LSESr of the current invention interferes with DHT production in the scalp. The DHT impacts the hair growth cycle in genetically susceptible individuals, causing the hair follicle to miniaturize, which results in hair thinning and eventually hair loss. Thus, the enhanced LSESr of the current invention inhibits 5α-Reductase 1 and 2 enzymes that convert testosterone to DHT, and thus, supports a healthy hair growth cycle, hair health, and becomes an important active ingredient to be used for supplements that help inhibit thinning hair.
The enhanced LSESr of the current invention has a high ratio of free fatty acids to total fatty acids that is greater than about 80.0% and enrichment of lauric, myristic, oleic and linoleic free fatty acids greater than about 82.0%, and inhibits 5α-Reductase types 1 and 2 enzymes that play an important role in androgenic hair thinning and eventually hair loss. The enhanced LSESr of the current invention interferes with DHT production and reduces the amount of DHT that may accumulate in the scalp, and which impacts the hair growth cycle. DHT may accumulate in the scalp and impact the hair growth cycle, causing the hair follicle to miniaturize in genetically susceptible individuals and result in hair thinning and eventually hair loss. The enhanced LSESr of the current invention inhibits 5α-Reductase type 1 and type 2 enzymes that convert testosterone to DHT and supports a healthy hair growth cycle and hair health. The enhanced LSESr may be an important active ingredient added to dietary supplements for addressing thinning hair and helping to make hair thicker. The enhanced LSESr of the current invention has a higher ratio of free fatty acids to total fatty acids as shown by experiments and will support hair thickness and hair fullness, and operate as a natural and safe ingredient for hair health.
It is possible to add vitamin D with the enhanced LSESr of the current invention. The vitamin D receptor expressed in dermal papilla cells will maintain hair follicle homeostasis, and especially anagen initiation. The dermal papilla cells are an important site for DHT impact on hair. The vitamin D receptor in both serums and tissues are significantly different in adults with androgenic hair loss than other controls (p=0). An insufficient amount of vitamin D may mean that keratinocytes in hair follicles are not able to regulate hair growth and shedding. It is known that women with mild-to-moderate female pattern hair loss (FPHL) have significantly higher mean serum vitamin D levels than women with severe hair loss. The micro-inflammation that does not lead to scarring of the scalp has been evident in women with FPHL and may be related to the hair follicle miniaturization process.
There are also synergies using the enhanced LSESr of the current invention with zinc because it has been shown to increase the proportion of follicles in anagen, hair volume, and hair appearance in men with AGA. Zinc deficiency may reduce the efficacy of Minoxidil treatment in men with AGA. Zinc may inhibit DHT production by limiting production of NADPH, which is necessary for the function of 5α-Reductase enzymes. A combination topical lotion containing the enhanced LSESr of the current invention and an herbal supplement containing zinc may be 50% more effective than either alone in male and female AGA. Some studies have shown that treatment with a topical saw palmetto combination product for four weeks increased the average hair count and terminal hair count at 12 and 24 weeks in 50 men with androgenic alopecia. The enhanced LSESr of the current invention will provide greater enhancements.
It also possible to formulate the enhanced LSESr of the current invention with rosemary oil, and optionally in combination with 5% Minoxidil. This combination should be significantly better than Minoxidil alone and improve the mean hair diameter and hair mass. The enhanced LSESr of the current invention may be combined with one or more of vitamin D, zinc, biotin, collagen, olive oil and rosemary oil to address androgenic-mediated hair thinning and loss, support hair growth and regrowth, and address micro-inflammation in the hair follicle in adults suffering from androgen-mediated hair loss. Zinc, which inhibits 5α-Reductase through a different mechanism of action than LSESr, may also act synergistically with the enhanced LSESr of the current invention to support hair follicle health, growth of new hair, hair volume and fullness, and overall hair health and appearance. The rosemary oil may act synergistically to support optimal hair growth in patients already using Minoxidil.
The enhanced LSESr of the current invention is produced using pre-extraction processing parameters for saw palmetto berries and specific processing parameters of ultra-high-pressure, supercritical CO2 extraction, which creates a hexane-like extract but without the harsh organic chemicals. It is rich in the free fatty acids which have good bioavailability and are absorbed twice as well as triglycerides, and includes better localization to the site of activity, such as the scalp or prostate as compared to triglycerides.
There now follows a description of parameters associated with the preprocessing of ripe saw palmetto berries as pre-extraction parameters, which in combination with specific processing parameters in the supercritical CO2 extraction, will produce the enhanced LSESr as in Experiments 5 and 6 in
In an example, dried mature saw palmetto berries of less than about 12% to about 13% moisture, and preferably ranging from 10% to 12% moisture, were released for the production process by a batch reference number. The saw palmetto berries by batch were introduced to a cryogenic milling system to create a consistent fine powder. The cryogenic milling system included a control feeder, and in an example, a 50 horsepower chopper mill with ½ inch screen openings. Different horsepower ratings may be used depending on the type of machine. A cryogenic conveyor included a liquid nitrogen addition to create chopped berries at about −40° C. In an example, the temperature was no greater than −20° C. This range can vary as much as 10-20 degrees (F. or C) above and below the −40° C. value.
Frozen material was dropped by gravity into a 50 horsepower pulverizing mill with a 3/16 inch screen opening. The feed rate was typically about 119+/−6 kilograms per hour (kg/hr) with a resulting mill amperage of about 4 amps, and an amperage range preferably of about 2.5 to 5.5 amps. The 50 horsepower chopper amperage had a similar amperage range and is in-line with the process at a consistent feed rate. The liquid nitrogen requirements were about 20+/−1 standard cubic feet (SCF) cryogenic nitrogen per kilogram of chopped and dried saw palmetto berries to achieve a recommended process temperature of feed into the mill on a continuous basis. This can vary by about 10% to 20% above and below these values.
The fine, milled saw palmetto powder about 100% less than about 4 mesh and about 95% less than about 12 mesh was fed into super sacks at about 600 to about 900 kilograms each on pallets. The entire lots once completed were stored by lot in a warehouse under hold, until aged to a target time. The aging was greater than about 15 days, and may preferably about 15 days to about 30 days, and in another example, be greater than 20 days, but not excessive such as beyond 40 day aging, and in an example, about 15 days to about 30 days for best results. The cryogenic milling followed by at least 15 days aging was found important with the CO2 extraction parameters. The completed milled and aged saw palmetto powder by lot was released to an extraction and separation process.
Referring now to
The aging impact on the free fatty acids (FFA) with fine milled berries as processed above was determined at a central Florida location having ambient warehouse conditions. An average temperature is shown in
Ambient aging conditions have an effect on the saw palmetto berries. Thus, various warehouse ambient conditions are important for the milled saw palmetto, and aging is appropriate prior to extraction.
The mature, dried, and milled saw palmetto berries that are aged are then extracted with the enhanced processing parameters. The scatter plot graph data shown in
Referring now to
Referring now to
In
The hold time impact on the variance of the percentage of free fatty acids and the fatty acids are shown in the test results as a bracketed line of
Different fatty acid statistical tables with ranges are shown in
This evidence also shows that the saw palmetto berries that are harvested and dried have various levels of fatty acids in a specific relationship to each other. The free fatty acid quality for the same fatty acids varies naturally over the season and region of harvest. By selecting specific berry preprocessing conditions and parameters and specific supercritical CO2 extraction parameters, it is possible to maximize and standardize to a tight variance the individual free fatty acid levels and their ratio relative to fatty acids that are important for prostate health and hair health. These results also add insight into the interactions and reactions occurring with the fatty acids and free fatty acids as based on the processing parameters and the post mill hold time.
Referring now to
The oleic acid weight fraction is shown in
The linolenic acid weight fraction versus the post milling hold time is shown in
There are probable interactions as one of the active fatty acids shift. An analysis is helpful about which fatty acid shifts the most and which fatty acids are important to the total free fatty acids and total fatty acids. The data may be analyzed by doing a regression analysis to determine which fatty acids and free fatty acids interact the most to the total free fatty acid in the process with the saw palmetto extract.
The table in
Referring now to the bar charts in
The graphs in
The multiple regression for the total free fatty acids and model building report are shown. The variables for maximizing total free fatty acids and the reactions and interaction of the fatty acids and free fatty acids are shown as they contribute to total free fatty acid and can be analyzed to determine any maximizing behavior. The five active fatty acids are grouped together as one factor (with free fatty acids as lauric, myristic, oleic, linoleic, and linolenic) and the volatile short chain fatty acids then analyzed. The R2 adjusted is 99.96% and the P-value is about 0.006 on the regression, which are very good values. The interactions and second order reactions with the shorter chains operating towards maximizing the total free acid.
The multiple regression for free fatty acids is shown in
The results of the various experiments, such as described relative to the data of
The fitted line plot of
Another scatter plot of the lauric acid and free fatty acid ratio versus the calculated percentage of oil in berries is shown in the graph of
Referring now to
Referring now to
Using the same data as above, the test for equal variance (
Referring now to
In this example, the process starts (Block 502) where 19, 958 kilogram (kg) of dried saw palmetto (Block 504) was fed through a feed hopper (Block 506). A magnet extracted any magnetic particles or contaminants (Block 508). The product was fed into the hammer mill (Block 510) where liquid nitrogen (Block 512) was applied at about −40° C. to about −30° C., which could possibly range from about −44° C. to about −27° C., a variation of about 108. The product was aged (Block 514) as noted above, up to about 15 days at least, and preferably at least greater than 15-20 days and up to about 15 to 30 days, or even 15 to 40 days. The temperature may vary during ageing as noted above with a desired mean of about 59° F. to about 78° F. As noted in the data above, aging to greater than 15 days has been found important together with the unique CO2 extraction parameters to optimize the contribution of the four bioactive free fatty acids.
Reference is made to some of the aging parameters described above that were followed. After aging, the aged and milled dried saw palmetto powder was placed into the supercritical CO2 extractor and carbon dioxide used as the supercritical fluid (Block 516) was applied at about 550 bar at 85° C. for CO2 extraction (Block 518). These values can range from about 495 bar to about 605 bar and 77° C. to about 94° C. In this example, based upon the 19958 kg initial amount, the spent marc for the solid waste leftover (Block 520) was obtained at about 16,645 kg.
The first fractionation (Block 522) occurred and the extract was separated at a first stage and the amount as the first separated extract (Block 524) was about 2, 426 kg. This first fractionation occurred at about 140 bar at about 77° C. in this example. This pressure could range from about 126 bar to about 350 bar, and the temperature range from about 35° C. to about 85° C. A second fractionation (Block 526) occurred at about 55 bar and at about 25° C. and the second separated extract (Block 528) was about 1, 164 kg. The pressure range may be about 30 bar to about 80 bar, and a temperature range may be about 12° C. to about 28° C. The first and second separated extracts may be dewatered and decanted (Block 530), followed by blending to form a final product of about 2, 377 kg (Block 532) corresponding to the enhanced LSESr. The process ends (Block 534).
It has been found that greater control over the saw palmetto berry selection and the aging process, such as in some cases aging close to 20 days based on berry selection, together with fine tuning of the supercritical CO2 extraction process of these selected aged berries, will enhance not only the free fatty acid profile, but also the four primary bioactive free fatty acids described above. This is advantageous because free fatty acids are the primary building blocks of hair as shown in the graphical representation of a hair follicle of
Table 2 below illustrates a comparison in the fatty acid and free fatty acid composition parameters of the new enhanced LSESr composition as compared to the original fatty acid and free fatty acid composition parameters as described above in the parent application filed on Aug. 8, 2022. The more enhanced compositional parameters are listed in the middle column for the new enhanced LSESr and the original enhanced LSESr compositional parameters described in the parent application are shown in the left-hand column. A third party is included for its fatty acid and free fatty acid compositional parameters in this table, which is a selected product commercially sold under the Dr. Best/Euromed product name as a saw palmetto extract. Both the original disclosed enhanced LSESr compositional parameters from the parent application filed on Aug. 8, 2022 and disclosed above, and more particularly, the new enhanced LSESr are produced using the supercritical CO2 extraction process with select aging of berries. The new enhanced LSESr with greater control over berry selection and aging and CO2 extraction has better fatty acid/free fatty acid parameters. The new enhanced LSESr is also referred to as USPlus Derm throughout this description.
In the following tables of
The bar chart in
The lower values of the compositional parameters for the USPlus shown with its example of Sample 1 compositional parameters in
Reference is again made to Table 2 below showing a comparison among the enhanced LSESr compositional parameters as described above and presented in the parent application filed Aug. 8, 2022 and the current new enhanced LSESr, and the competition compositional parameters, such as Dr. Best/Euromed products. The new enhanced LSESr has been tested and found more effective as explained below. Most notable improvements for the new enhanced LSESr shown in Table 2 are seen with the larger percentage of free fatty acids to total fatty acids, the higher percentage of LA and LNA, the much higher percentage of free fatty acids, and importantly the higher percentage of the four bioactive free fatty acids (FFA4) and FFA4 to TFA. This also allows a smaller dose of 70 mg to 320 mg to be used instead of 200 mg to 320 mg.
The benefits of the new enhanced LSESr as the USPlus Derm have been determined from a series of experiments, including a) in vitro, b) ex vivo, and c) topical work. The beneficial results were obtained from the experimental work conducted by QMA Monasterium GMBH using samples of the new enhanced LSESr as the USPlus Derm, including a topical serum study with different volunteer participants over a test period of three months as explained below.
It has been found that treatment with the new enhanced LSESr, such as disclosed in Table 2, also referred to as the USPlus Derm, prolongs anagen through positive modulation of dermal papilla inductivity, and reinforces the hair follicle stem cell niche in organ cultured, occipital healthy male scalp hair follicles.
Male pattern androgenetic alopecia areata (mpAGA) is driven by an increased 5-α reductase (5-αR) activity in hair follicles (HFs) of affected frontal-vertex scalp skin areas. The supercritical CO2 extraction process of aged saw palmetto berries as described above, including the saw palmetto berry preparation, allows production of the new enhanced LSESr as USPlus Derm. The new enhanced LSESr exerts not only 5-αR inhibitory activity, but it also contains more concentrated levels of the bioactive free fatty acids shown in Table 2 compared to other saw palmetto extracts. As integral lipids are important components of the hair shaft and regulator of the hair cycle, the clinical investigation determined whether the new enhanced LSESr as the USPlus Derm may have enhanced hair growth promoting effects, which are independent of 5-αR activity.
Experiments were conducted that used micro-dissected full-length occipital hair follicles (HFs) from five male healthy donors and applied the new enhanced LSESr as USPlus Derm at two different concentrations (i.e., 0.4 and 10 μg/ml). The 10 μg/ml showed 5-αR inhibition in vitro. Anagen was prolonged by both concentrations, but only 0.4 μg/ml of the new enhanced LSESr as USPlus Derm tendentially increased proliferation of hair matrix keratinocytes (Ki-67+ cells).
As shown in
The graph at
The bar chart in
The dermal papilla (DP) inductivity was determined by examining alkaline phosphatase activity and versican expression to assess the mechanism underlying anagen prolongation. The application of 0.4 μg/ml of the new enhanced LSESr known as USPlus Derm tended to increase both parameters. The emigration of dermal papilla (DP) fibroblast into the DP stalk is one of the first signs of catagen initiation and, if persistent, leads to reduced DP size and hair follicle (HF) miniaturization. Treatment with the enhanced LSESr as the USPlus Derm enhanced DP size and significantly reduced cell density in the DP stalk, such as at 10 μg/ml, whereas the DP cell number was unaffected, indicating reduced fibroblast emigration in the presence of USPlus Derm.
Lastly, expression and proliferation of K15+ HF stem cells (HFSCs) were analyzed following the treatment from the new enhanced LSESr as the USPlus Derm treatment. Application of 0.4 μg/ml of the new enhanced LSESr as the USPlus Derm enhanced K15 expression and reduced the number of Ki-67+K15+ cells, while K15+ cell numbers were unaltered, indicating HF stem cell niche reinforcement. Taken together, this data shows 5-αR independent, beneficial effects, i.e., enhanced hair growth and HFSC stemness, of the new enhanced LSESr as the USPlus Derm in “clinically” healthy HFs and support its further investigation for the management of male pattern hair loss and other, 5-αR independent, hair growth disorders.
The parameters that were evaluated include hair follicle elongation (hair shaft production) as an ex vivo experiment using a digital brightfield microscope. The methodology included a) embedding hair follicles for in situ protein analysis, b) collecting medium for follow-up analysis, c) hair follicle sectioning, d) microscopic hair cycle staging and scoring using Ki-67/TUNEL immunofluorescence and Masson Fontana histochemistry, e) analyzing hair matrix proliferation and apoptosis using Ki-67/TUNEL immunofluorescence, and f) analyzing melanin clumping in the pigmentary units using Masson Fontana histochemistry.
To determine the hair follicle length, each hair follicle was measured from the end of the bulb connective tissue sheath to the end of the hair shaft using a digital light microscope at 50× magnification, such as a VHX900 microscope from Keyence Corporation, Osaka, Japan and affiliated software.
Ectopic melanin clumps were counted in Masson Fontana staining of hair follicles at the end of the culture in a defined region of interest, starting at the Auber's line and including the precortical hair matrix. Incontinence and melanin clumping were signs of hair follicle cytotoxicity and dystrophy.
Hair cycle staging was performed at the end of the culture and determined according to several established macroscopic and microscopic parameters. HFs were microscopically evaluated for the hair cycle staging using Masson-Fontana histochemistry and Ki-67/TUNEL immunostaining as previously described. To each hair cycle stage, an arbitrary score was given as following: a) anagen: 100; b) early catagen: 200; c) mid catagen: 300; d) late catagen: 400; and e) dystrophic: 500.
The quantitative evaluation of hair matrix keratinocytes proliferation and apoptosis was performed on images of Ki-67/TUNEL stained hair follicle cryosections. Initially, the hair follicle region of interests as germinal hair matrix (gHM1+gHM2) and precortical hair matrix (pcHM) were predicted by leveraging a transfer learning approach from AI techniques. The accuracy and correctness of ROIs recognition was verified by a human investigator using an ImageJ based (in-house implemented) QC pipeline. In a second AI phase, DAPI positive cells were segmented within the predicted ROIs by using the Cellpose open-source AI library on DAPI channel images.
The subsequent counting of the total number of predicted DAPI positive cells in DAPI channel images was automated using a Python programming language based script. Next, in each predicted DAPI positive cell, the mean intensity was computationally determined in the corresponding TRITC channel (Ki-67) image and a threshold based analysis (filtering) was implemented to identify and count Ki-67 positive nuclei. In a final step, the germinative hair matrix keratinocyte proliferation was calculated as the percentage of Ki-67 positive nuclei in gHM (1+2).
The quantitative assessment of hair matric keratinocytes apoptosis was performed manually in the FITC channel in AI predicted ROIS (gHM1, gMH2) and expressed as the percentage of TUNEL (TdT-mediated dUTP-biotin nick-end labeling) positive nuclei in gHM and pcHM per DAPI positive nuclei (AI counted).
Statistical analyses was performed using Graphpad Prism 9 (GraphPad Software Inc.). Data was tested for normality distribution suing the D'Agostino and Pearson omnibus normality test before subsequent analysis using the appropriate parametric (One-way ANOVA) and non-parametric (Kruskal-Wallis) tests, including multiple comparison testing. Two-column comparisons were performed using Mann-Whitney U-test whenever the data did not follow a Gaussian distribution or an unpaired student's t-test whenever they did. Data was expressed as mean±SEM or fold-change of mean±SEM. All data was analyzed by comparing the different conditions to the vehicle and p-values <0.05 were regarded as significant. The significance was noted as follows:
Further details of the tests such as bar charts and images are now explained.
As shown
The hair cycle analysis is shown in
These experimental results show that the new enhanced LSESr as the USPlus Derm promotes anagen, tendentially enhances dermal papilla inductivity, and maintains the hair follicle stem cell niche. The majority of those effects may be seen with the lower concentration of the new enhanced LSESr at 0.4 mcg/ml. As also evident, there are interesting effects on the 5 alpha-reductase-inhibition-independent hair growth-promoting effects.
Referring now to Table 3 below, there is illustrated a topical serum formulation as a lotion that includes a 2% LSESr incorporating the new enhanced LSESr as the US Plus Derm in a human consumer use trial. The percentages, such as w/w shown in Table 3, are specific to the tested sample used by the consumers as the participants. The ranges of those values may vary. For example, the deionized water may vary from about 72.00% to about 80.50% and the hydroxyethyl urea may vary from about 4.75% to about 5.25%. The hydroxyethyl urea moisturizes and hydrates and operates as a hydrophilic and moisture-retaining component. It also operates as a humectant. The acacia senegal gum and xanthan gum may vary from 0.28% to 0.32% in non-limiting examples and operate as a thickening, stabilizing, and texturizing component. The acacia gum may be used as an emulsifier while the xanthan gum may thicken aqueous solutions. It may operate to thicken the topical serum.
The new enhanced LSESr as USPlus Derm may vary from 1.9% to about 2.1% in this non-limiting example, but the ranges may vary more. Polysorbate 20 may be used and vary from about 5.7% to about 6.3%. Polysorbate 20 is a non-ionic surfactant that operates as a wetting agent and a washing agent, stabilizer, and solubilizer, and an excipient that may stabilize an emulsion and suspension. As is well known, it is made by ethoxylating sorbitan monolaurate as 20 repeated units of polyethylene glycol. The hydrophilic groups are the polyethers known as the polyoxyethylene groups that are polymers of ethylene oxide. The numeric designation for 20 refers to the lipophilic group as the oleic acid. Some prior art commercial formulations of saw palmetto extracts use higher amounts of polysorbate 20, but the 5.7% to 6.3% has been found adequate with one reason out of several that the new enhanced LSESr includes that higher percentage of oleic acid as a free fatty acid.
The denatured alcohol may vary from 9.5% to about 10.5%. A fragrance may be added and vary from about 0.09% to about 0.11%, while citric acid may vary from about 0.04% to about 0.06% and help the pH maintain to about 6.0 to about 6.5 for the formulation as a serum/lotion. The citric acid may aid in bringing pH levels down to improve hair appearance and manageability by reducing frizz.
The clinical outcome of the topical application of the hair growth composition, i.e., the 2.0% new enhanced LSESr as USPlus Derm, in men and women shows its positive benefits. The clinical study was performed to determine the benefits of the composition as a topical hair serum as a blinded consumer user trial. As background, the hair loss treatment products market was valued at about US $20.5 billion in 2023 and is projected to reach a market size of about US $32.92 billion by the end of 2030. Over the forecast period of 2024-2030, the market is projected to grow at a compound annual growth rate of about 7%. People can typically lose 50 to 100 hairs a day, but this usually is not noticeable because the same number of new hairs begin growth every day.
Noticeable hair loss occurs when new hair does not replace the hair that has fallen out, or grows in as miniaturized, thinner hair. Hair loss is typically related to one or more of the following factors: genetics, hormonal changes, underlying health issues or medication use, stress, or damage to hair follicles, including chemical treatment. Hair loss can be reversed by drugs like finasteride, which may also be used for treating benign prostate hyperplasia since it is a 5-α reductase inhibitor, but many consumers instead are looking for cost-effective non-therapeutics as a natural solution to address the problem.
The new enhanced LSESr as USPlus Derm is highly enriched in free fatty acids important for hair follicles and the hair growth cycle. It includes a naturally concentrated blend of the four most important bioactive free fatty acids indicated and described above, which are important to address hair loss and support hair regrowth. They are also important in improving prostate health.
The aim of the blinded consumer use trial was to investigate the outcome of a 2% topical application as a serum formulation of the new enhanced LSESr as USPlus Derm in adult men and women who had been experiencing unresolved hair loss for at least about six months.
Study Design: The clinical study was conducted with 15 men and 7 women, aged 25 to 65 (average 49.3+11.0 years) who had been experiencing unresolved hair loss for at least 6 months. The pie chart in
The blinded clinical study product, i.e., a 2% serum of the new enhanced LSESr as USPlus Derm topical serum, was applied using two (2) pipettes of product 3-times a week on wet hair and once a week on dry hair and massaged into the scalp. This 2% serum was formulated to be used as a leave-in product that did not require washing off. Acceptance of the sensory characteristics of the product and tolerance/safety were also monitored.
In addition to concerns for hair loss, other hair problems were noted by the study subjects at baseline. Over 40% of study subjects were suffering from dandruff, and over 30% suffered from damaged, brittle or fragile hair. The 22 study participants had a wide range of hair types (long, short, straight, wavy, etc.) and hair conditions (oily, dry, etc.) at study entry.
Study participants completed a 15-question closed assessment on hair perception and hair characteristics at baseline and each month for 3 months. Results were reported as the percent of study participants who marked top-2 box preference, Strongly Agree or Agree, for each question and compared to baseline. Study participants also completed a numeric ranking questionnaire on general hair loss, hair loss with styling, and hair density at these time-points. For 10-point numeric ranking, outcome was 0=No hair loss to 10=Severe hair loss for General Hair Loss and for the question on Hair Loss with Styling; Hair Density was scored 0=Very low hair density to 10=Very high hair density. Numeric rank questionnaires were self-reported at baseline and at monthly intervals for 3 months. In addition, weekly status of perception of hair loss was collected in weekly phone calls with study participants at the same time that tolerance/safety reporting was documented.
Results: After 2 weeks of use of the 2% new enhanced LSESr as the USPlus Derm topical serum, 27% of study subjects noted less hair loss, and 50% of study participants reported a favorable improvement after 3 weeks. After 6 weeks of use, the improvement in satisfaction in hair loss was statistically significant.
At 4 weeks, 60% of study participants noticed the 2% new enhanced LSESr as USPlus Derm topical serum appeared to reduce hair loss and that they were losing less hair every day. 68.2% of the study participants reported that their hair had more texture and that the product seemed to stimulate their scalp. Also, 63.6% of the participants agreed or strongly agreed that their hair seemed strengthened, they felt like they were losing less hair when styling, and that the product helped maintain better hair density. Within the first month of use, 59.1% of the study participants said the clinical product appeared to stimulate hair growth/regrowth.
After 8 weeks of topical application of the new enhanced LSESr as the USPlus Derm topical serum, the results were improved and findings on hair loss were significantly changed from baseline. 80% of study participants felt they were losing less hair every day and 70% reported that the product seemed to reduce their hair loss. On a numeric scale, hair loss in general had decreased by 18.7%. Hair loss when styling decreased by 29.0% after 2 months of product use and was a significant change from baseline. Hair density had increased by 10.2% and was tangentially significant.
After 3 months of use of the 2% new enhanced LSESr as the USPlus Derm topical serum, the effectiveness on hair loss increased further and reached very good results, according to the study investigators. Overall, 90.9% of study participants stated that the product helped maintain better hair density and 86.4% of participants reported that the product seemed to reduce hair loss. Also, more than 75% of the study participants agreed that they appeared to be losing less hair every day. When styling, the participants reported that the product appeared to stimulate their scalp. For 72.7% of the study participants, after 12 weeks of the product application, they felt that the product appeared to stimulate hair growth/regrowth and that the areas affected by hair loss seem less visible. These results were significant.
At the end of the 12-week consumer use clinical trial, numeric scoring on questionnaires indicated that participants felt hair loss in general had decreased by 22.48. Hair loss when styling decreased by 34.2% and was significant. Also, hair density increased by 19.5% and was also significant.
Dermatological assessments at 12 weeks found significant improvement in hair loss for both men (8.7%, p=0.009) and women (25.2%, p=0.004). The presence of dandruff was visibly less on dermatological assessment, decreasing by 31.7%.
Conclusions: Consumers saw a difference in hair density and/or hair loss as early as 2 weeks (27%) and 50% saw an improvement by 3 weeks. After 6 weeks of use the improvement in satisfaction in hair loss was statistically significant.
Hair loss when styling was statistically significantly improved by week 8, and further improvement noticed at week 12. Hair density improved significantly from baseline at week 8, and further improvement in hair density to about 20% by 3 months. The dermatological change in hair loss score from baseline was statistically improved for both men and women, and scalp/skin improvement in reduction of participants suffering from dandruff was a key finding.
In addition to hair loss, study participants saw an improvement in hair appearance, such as its suppleness or manageability. Participants also noted the improvement in dandruff, and it was significant. At 12 weeks, study participants felt that the blinded product as the 2% new enhanced LSESr as the USPlus Derm topical serum was able to significantly impact (stimulate) hair regrowth, as well as improve the visible look of hair loss on the scalp.
As noted before, free fatty acids are important building blocks of hair, supporting hair characteristics and shine, and play an important role in keeping hair in growth and re-balancing the hair growth cycle. The new enhanced LSESr as the USPlus Derm topical serum is highly enriched in free fatty acids and includes the concentrated blend of the four most bioactive free fatty acids noted above that are known to drive hair growth and the hair growth cycle. The topical serum was able to give men and women visible benefits to their hair and scalp in this blinded consumer use trial.
Study participants started noticing a difference after 3 weeks, and the results were statistically improved from baseline after 6 weeks. General hair loss and hair loss when styling improved at each monthly assessment, as did an improvement in hair density. Hair loss when styling and hair density were significantly improved at 8 weeks and further improved by 12 weeks. Finally, official dermatological assessment noted statistically significant improvement in hair loss scoring for both men and women, as well as an important reduction in dandruff.
This consumer use clinical trial demonstrated the benefit of the 2% new enhanced LSESr as the USPlus Derm topical serum and the ingredients' concentrated bioactive free fatty acids to improve hair characteristics, address hair loss and hair regrowth and scalp conditions, such as dandruff. The product was very well tolerated. The sensory characteristics of the product were well accepted by the study participants. The new enhanced LSEsr as USPlus Derm is an important new ingredient for men and women looking for a natural solution to maintaining hair, addressing hair loss, improving hair density and supporting healthy hair and scalp.
Reference is made to the Table 4 below for the data comparison among: a) the 2% new enhanced LSESr formulated as the topical serum, b) an existing 20% VISPO topical lotion, and c) 100 mg of VISPO per 400 mg capsule.
The VISPO topical lotion includes a standardized saw palmetto extract that contains about 2% to about 3% beta-sitosterol and about 85% total fatty acids. Each 4 mg capsule contains about 100 mg of the VISPO. The composition of the VISPO product with other ingredients is shown in Table 5 below as taken from the article by Sudeep et al., Oral and Topical Administration of a Standardized Saw Palmetto Oil Reduces Hair Fall and Improves the Hair Growth in Androgenic Alopecia Subjects-A 16-Week Randomized, Placebo-Controlled Study; Clinical, Cosmetic and Investigational Dermatology, 2023:16, pp. 3251-3266. The placebo composition is also illustrated in Table 5 in that study.
The data comparison as shown in Table 5 for VISPO topical helps explain the advantageous benefits of the 2% new enhanced LSESr as USPlus Derm as the topical serum as an improvement over competitor products such as the VISPO topical constituting 20% w/w of the Serenoa repens oil and VISPO oral product.
The Sudeep et al. sample size calculations are explained below.
Sample size calculation: The sample size calculation was based on the difference of two treatments considered to be medically relevant. Assuming a common standard deviation of 2.695 at the end of treatment, 18 per group would be sufficient to detect a difference of 1.89 in mean difference between the two treatments with power of 80% and a 0.05. two-sided level of significance.
A total of N number of subjects is required at each Treatment group in the end of the study with all the data being complete for analysis, but a proportion (q) are expected to drop out before the study ends. In this case, the following total number of subjects (N1) would have to be enrolled to ensure that the final sample size (N) in each Treatment group is:
Where q is the proportion of attrition and is generally 10% in this type of study, we considered 10%.
The benefits of the new enhanced LSESr as USPlus Derm are readily apparent from the various studies discussed above.
The composition may include one or more of minoxidil, finasteride, and dutasteride formulated for topical or oral use. These three ingredients may work together with the new enhanced LSESr because when used together to treat hair loss, the three components of the minoxidil, finasteride, and dutasteride may treat hair loss by working through different mechanisms to target the root causes of the hair loss and lead to better results when combined.
For example, the finasteride and dutasteride primarily reduce the effects of dihydrotestosterone (DHT) on hair follicles by inhibiting its production. Minoxidil, on the other hand, works by stimulating hair growth directly and improving blood flow to the scalp. Finasteride and dutasteride are 5 alpha-reductase inhibitors and block the conversion of testosterone to DHT, while minoxidil acts as a vasodilator. They work in a synergistic effect because they address different aspects of the hair loss process. Minoxidil may operate when the sulfotransferase enzyme in the human scalp converts the minoxidil into minoxidil sulfate, which is the active form of the molecule. Minoxidil shortens the telogen phase of the hair growth cycle, which is the resting phase, causing the hair follicles to enter into the growth phase sooner. Minoxidil also has an antihypertensive effect by opening the adenosine triphosphate (ATP)-sensitive potassium channels to relax the smooth muscle vasculature and reduce peripheral resistance to help lower blood pressure levels.
Because the finasteride and dutasteride prevent the testosterone from turning into the DHT, they can operate together to alleviate benign prostate enlargement and also help shrink the prostate and reduce hair loss. The finasteride as a 5 alpha-reductase inhibitor is in antiandrogen, and in some formulations, may decrease the production of DHT by about 70%. Taking the finasteride leads to a reduction in scalp and serum DHT levels and maintains or increases the amount of terminal hairs in the anagen phase by inhibiting and sometimes reversing miniaturization of the hair follicle.
Finasteride may be administered with an oral dosage of about 5 mg/day and 1 or 5 mg/day for prostate applications and even hair dosage. 0.2 mg/day has been found to suppress DHT levels. However, finasteride may lack certain inhibitory effect on the 5 alpha-reductase Type I isoenzyme as compared to Type II. Dutasteride, on the other hand, was found to be an inhibitor of all three isoenzymes of 5 alpha-reductase, which can reduce DHT levels in the entire body by more than 99%.
The new enhanced LSESr has potential use in compounding pharmacies or formulations where the formulation may have a reduced amount of one or more of the Minoxidil, Finasteride, and Dutasteride, whether formulated for topical or oral use. This is beneficial. Often 5% Minoxidil is used in current applications, but with the combination with the new enhanced LSESr, lesser amounts may be used, e.g., 2% to 3% as a non-limiting example. The same applies to use of the new enhanced LSESr whether with one or more of Minoxidil, Finasteride, and Dutasteride are applied in a composition or other formulation, whether topical or oral.
As noted before, any one or more of zinc, vitamin D, biotin, collagen, rosemary oil and olive oil may be added. The collagen may help with hair growth by providing amino acids that support hair structure and keratin production. Collagen may also help fight free radical damage that can cause hair to age or become gray. Collagen helps hair growth via its amino acids, and provide structural support, fight free radical damage and slow thinning. For example, collagen contains amino acids like proline, glycine, and hydroxyproline. These amino acids are used to build keratin, the protein that makes up hair. Collagen provides structural support to hair and scalp and can help fight free radical damage caused by pollution, UV rays, inflammation, and aging. Collagen may help slow down hair loss. Supplementation with collagen peptides has the potential to support hair growth and healthy hair.
Collagen peptides may also maintain hair follicle stem cell quiescence ex vivo. Collagen peptides preserve the populations of human hair follicle stem cells and their progeny ex vivo. Collagen works with hyaluronic acid to hydrate and repair the dermis and follicle environment. Collagen supplements can stimulate collagen synthesis in the scalp, which may improve blood supply to hair. Collagen's antioxidant properties may help combat damage from free radicals, which could negatively affect hair follicles. Although different forms of collagen may be used, it has been found that hydrolyzed collagen peptides can be advantageous. This form of collagen is more easily absorbed by the body and may be more effective for hair growth. Amounts can vary, but it is possible to add about 2.5 to 15 grams per day for oral consumption, including topical administration.
It has also been found that biotin is helpful for healthy hair growth. Biotin is a coenzyme for carboxylase enzymes that assist various metabolic reactions involved in fatty acid synthesis, branched-chain amino acid catabolism, and gluconeogenesis important for maintenance of healthy skin and hair. The biotin may be administered at about 3 milligrams (or 3,000 mcg). This amount can vary depending on individual needs.
Many modifications and other embodiments of the invention will come to the mind of one skilled in the art having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the invention is not to be limited to the specific embodiments disclosed, and that the modifications and embodiments are intended to be included within the scope of the dependent claims.
This continuation-in-part application is based upon U.S. patent application Ser. No. 17/818,011 filed Aug. 8, 2022, which is based upon U.S. provisional Application No. 63/234,308 filed Aug. 18, 2021, the disclosures which are hereby incorporated by reference in their entirety.
| Number | Date | Country | |
|---|---|---|---|
| 63234308 | Aug 2021 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 17818011 | Aug 2022 | US |
| Child | 19049105 | US |
