Non-aluminum-based hidroschesis and body odor inhibition composition

Abstract

The invention discloses a composition which does not contain aluminum and has the effects of reducing sweat secretion and inhibiting bromhidrosis. High-energy leukocyte extract, chitosan oligosaccharide, chitosan, vitamin B6 and curcumin are adopted as main active ingredients, and transfer of bromhidrosis precursor substances to the body surface is weakened by reducing sweat secretion; further, proliferation of body odor bacteria is selectively inhibited through colonization of probiotics, so that conversion of odor substances is reduced, and finally, the purpose of inhibiting bromhidrosis is achieved. The aluminum-free composition disclosed by the invention is clear in component, high in biological safety, high in compatibility, controllable in quality, good in batch-to-batch stability and high in safety, and all components are listed in CFDA (Cosmetic New Raw Material Registration Registration Registration Registration Registration Registration Registration Registration Registration Registration Registration Registration).

Description

Technical Field

[0001] The invention belongs to the field of cell and bioengineering technology, and particularly relates to a non-aluminum-based antiperspirant and body odor-inhibiting composition. Background Art

[0002] The prevalence of bromhidrosis in my country is approximately 4-6%, with a peak incidence during adolescence. It poses a significant psychological and social burden. The sweat of people with bromhidrosis is rich in fatty acids, which are broken down by bacteria on the body's surface, producing a distinctive odor. Due to differences in the Abcc11 gene, apolipoprotein D (ApoD) binds to the unsaturated fatty acid E-3M2H, which is secreted and transported to the body by apocrine sweat glands in the axillae. This secretion is broken down by bacteria on the skin's surface, producing a distinctive, pungent odor. The specific mechanism of bromhidrosis is not fully understood. Overactive apocrine sweat glands accelerate sweat secretion, exacerbating bromhidrosis. Local inflammatory diseases can contribute to this overactivity, while localized microinflammatory reactions triggered by sub-health conditions (such as high oxidative stress) are more common, manifesting as increased expression of inflammatory proteins in sweat gland cells. The main bacterial species responsible for body odor are Staphylococcus epidermidis and Corynebacterium xerosis. The precursors of axillary odor in sweat are odorless. After being decomposed by bacteria, they release odorous substances, such as E-3-methyl-2-hexenoic acid (E-3M2H). Killing or inhibiting the growth of these bacteria can effectively inhibit the occurrence of axillary odor.

[0003] Severe cases of bromhidrosis require surgical removal of the apocrine glands under the arms to reduce sweat secretion for treatment. Most mild cases can significantly alleviate symptoms by using antiperspirants, regularly washing to remove secretions, and reducing bacterial growth. Aluminum salt antiperspirants form a gel-like substance with sweat on the body surface, reducing sweat flow; aluminum salts themselves are also antibacterial ingredients, preventing bacterial metabolism from producing odorous products. Under normal physiological conditions, sweat forms a protective lipid film with sebum on the body surface; however, excessive use of aluminum-based antiperspirants can clog pores and hinder the excretion of metabolites. The biocompatibility and safety of aluminum-based products have long been questioned. Therefore, the development of non-aluminum-based antiperspirants and deodorizing products meets the latest needs of the cosmetics industry. Summary of the Invention

[0004] In response to the market demand for existing antiperspirant and deodorant cosmetic products and the shortcomings of existing technologies, the present invention provides a composition that does not contain aluminum components and has synergistic antiperspirant and axillary odor inhibition effects; by using raw materials included in the CFDA "Catalogue of New Cosmetic Ingredients", the product's biosafety and compatibility are improved; the defects of existing aluminum-based products are completely eliminated, meeting the demand for high-end biological cosmetic raw materials.

[0005] Specifically, the present invention provides a non-aluminum-based antiperspirant and body odor-inhibiting composition (abbreviated as Deswodor-Lac), comprising the following components in parts by weight: 5-20 parts of chitosan dry powder, 20-40 parts of chitosan oligosaccharide dry powder, 0.05-0.5 parts of high-energy leukocyte extract freeze-dried powder, 2-8 parts of vitamin B6, and 2-8 parts of probiotic freeze-dried powder.

[0006] The preferred non-aluminum-based antiperspirant and body odor-inhibiting composition of the present invention comprises the following components in parts by weight: 10 parts of chitosan dry powder, 35 parts of chitosan oligosaccharide dry powder, 0.1 parts of high-energy leukocyte extract freeze-dried powder, 5 parts of vitamin B6, and 5 parts of probiotic freeze-dried powder.

[0007] The high-energy leukocyte extract of the present invention is obtained through three steps: the first step is to separate the leukocytes in the blood of slaughtered animals, the second step is to perform stress removal, and the third step is to crush and extract the contents.

[0008] Specifically, the second step of destress treatment uses a destress treatment solution, which is composed of 0.9% NaCl, 1% hydroxyethyl starch, 0.125% taurine, 0.325% sodium ascorbyl phosphate, 1.5% ectoine, 1% proline, and 2% sodium butyrate in percentage by mass. The destressing is completed at 32°C, 5% CO2, and a volume ratio of the treatment solution to the white blood cell suspension of 10:1 at 4-6 hours.

[0009] The slaughtered animals can be rabbits, horses, cattle, sheep, deer, pigs and other farmed animals.

[0010] The preferred probiotic freeze-dried powder of the present invention is one or more of Lactobacillus bulgaricus freeze-dried powder, Lactobacillus delbrueckii family freeze-dried powder, probiotic yeast freeze-dried powder, and Bifidobacterium freeze-dried powder.

[0011] A further preferred concentration of the lyophilized Lactobacillus bulgaricus powder is 10 billion CFU / g.

[0012] The composition of the present invention can be prepared as dry powder, aqueous solution, emulsion or colloid.

[0013] The present invention provides a method for obtaining a high-energy liquid leukocyte extract by destressing. Leukocytes of slaughtered animals are separated and obtained at 4°C; they are destressed by incubation at a sub-low temperature (32°C) using a special destressing treatment solution; and high-energy leukocyte extracts are obtained according to conventional steps. After destressing, the key functional protein indicators of the leukocyte extract are improved. In the present invention, the use of high-energy leukocyte extracts has two purposes: (1) improving the inflammatory environment of skin cells in the axillary region; and (2) high-energy leukocytes are effective microbial inhibitors.

[0014] In the present invention, the chitosan-chitosan oligosaccharide mixture is used in the special environment where sweat and armpit odor occur. (1) Chitosan-chitosan oligosaccharide powder, the solubility of chitosan is extremely low, causing pores to be blocked and closed, and the secretion of sweat to the skin surface is reduced, but this blockage is temporary and reversible. The pores reopen as the powder is washed off during bathing; compared with aluminum-based antiperspirants, this "blocking" method is milder. (2) The pH value of sweat is about 4-5. At this time, the chitosan partially dissolves to form a membrane colloid. The water retention of the colloid helps to maintain the effective ingredients for a long time. (3) Chitosan-chitosan oligosaccharide is a good biological antibacterial component. Blending enhances bioavailability and complements each other in solubility, absorption and mechanism.

[0015] The vitamin B6 in the present invention is used to improve the excessive oil secretion of the skin.

[0016] Curcumin in the present invention is used to improve the skin environment of the armpit, reduce the micro-inflammatory reaction caused by excessive generation of free radicals, and further weaken the activity of apocrine sweat gland cells.

[0017] The use of probiotics (preferably Lactobacillus bulgaricus) in the present invention is suitable for the special environment where sweat and axillary odor occur. (1) As intestinal probiotics, safety is guaranteed; and they can colonize on the surface of animal skin. (2) The temperature, pH, and oxygen-rich environment of the axilla are suitable for the growth of lactobacilli. (3) Chitosan-oligochitosan not only does not significantly inhibit the growth of lactobacilli, but can also act as a prebiotic to promote the improvement of the bacterial flora. (4) During the oxygen-rich growth process, low concentrations of the aromatic substance acetaldehyde can be produced, which can further improve body odor.

[0018] Beneficial effects of the present invention:

[0019] The composition of the present invention can reversibly and temporarily block the secretion channels of sweat glands, inhibit the secretion of apocrine sweat glands in the armpits, and reduce the transport of sweat to the body surface; through active biological preparations (high-energy leukocyte extracts), it can weaken the local inflammatory response of skin cells in the armpits, reduce cell activity, and reduce the production of precursor substances of underarm odor; utilize multiple biological antibacterial components to regulate the underarm microbial community, selectively inhibit or kill bacteria that transform underarm odor; and regulate the underarm microbial flora through the colonization of probiotics on the body surface. The synergistic effect of various components can achieve the purpose of inhibiting underarm odor. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The stress index of the high-energy leukocyte extract is improved by the de-stressing treatment in Example 1 of the present application;

[0021] Figure 2 The de-stressing treatment in Example 1 of the present application improves the inflammatory indexes of the high-energy leukocyte extract;

[0022] Figure 3The selective inhibition of the axillary odor transforming strain and the probiotic strain in Deswodor in Example 3 of the present application;

[0023] Figure 4 The effect of Deswodor on the growth state of sweat gland cells in Example 4 of the present application;

[0024] Figure 5 The effect of Deswodor on sweat gland cell viability in Example 4 of this application;

[0025] Figure 6 This is a comparison of the underarm odor occurrence indicators of normal sweat gland cells and underarm odor sweat gland cells in Example 4 of the present application;

[0026] Figure 7 These are the indicators related to the occurrence of underarm odor in underarm sweat gland cells that Deswodor improves in Example 4 of the present application. DETAILED DESCRIPTION

[0027] In order to make the experimental purpose, technical solutions and advantages of the present invention more clear, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention, but are not intended to limit the present invention.

[0028] Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional in the art. Experimental methods in the following examples, where specific experimental conditions are not specified, are generally performed under conventional experimental conditions or according to the experimental conditions recommended by the manufacturer. Unless otherwise specified, the reagents and raw materials used in the present invention are commercially available.

[0029] Example 1: Obtaining high-energy leukocyte extract

[0030] 1) Separate the blood of slaughtered rabbits by standard density gradient centrifugation to obtain rabbit peripheral blood leukocytes, and adjust the cell density to 5×10 7 ~1×10 8 / mL, sampling.

[0031] 2) Prepare a leukocyte destressing solution by mass percentage: 0.9% NaCl solution containing hydroxyethyl starch 130 (1%), dextran 40 (2%), taurine (0.125%), sodium ascorbyl phosphate (0.325%), ectoine (1.5%), proline (1%), and sodium butyrate (2%). After fully dissolving, sterilize and filter through a 0.22 μm filter membrane.

[0032] 3) Under sterile conditions, the treatment solution and cells were mixed at a volume ratio of 10:1, and the cell concentration was approximately 5×10 6 ~1×10 7pieces / mL.

[0033] 4) Induce and incubate the immune cells at 32° C., 5% CO 2 , and saturated humidity for 4-6 hours.

[0034] 5) Wash with 0.9% NaCl solution at 4°C at 3000 rpm to obtain stressed leukocytes and take samples.

[0035] 6) Obtain high-energy leukocyte extracts according to standard leukocyte extract preparation methods (ultrasonic disruption, centrifugation to remove cell debris, and 3KD ultrafiltration to remove small molecules), and take samples to test corresponding biological indicators.

[0036] 7) A freeze-drying method is used to obtain a high-energy leukocyte extract dry powder.

[0037] 8) Detection of stress marker levels: The heat stress protein family (HSP family) is a cell stress marker. Approximately 5 million cells were collected before and after de-stressing, and the changes in HSP70 transcription levels were compared using an RT-PCR kit, with GAPDH as the internal reference. In this example, freshly isolated leukocytes were in a high stress state (untreated - 0 hr); de-stressing treatment (treatment solution - 32°C - 4 hr) significantly reduced the stress level; in the absence of treatment solution, the expression of stress markers remained essentially unchanged after 4°C treatment (4°C - 4 hr), while sub-low temperature incubation resulted in limited recovery of stress (32°C - 4 hr). The results are shown in Table 1. Figure 1 shown.

[0038] 9) Inflammatory marker detection: Compare the changes in the transcription levels of IL-1β (pro-inflammatory factor indicator) and IL-10 (inflammatory inhibitor) before and after de-stress treatment. In freshly isolated white blood cells, the pro-inflammatory factor IL-1β index is at a high level; de-stress treatment (treatment fluid -32℃-4hr) can significantly improve the outcome; in the absence of treatment fluid, the level of inflammatory markers remains basically unchanged after treatment at 4℃ (4℃-4hr), and mild hypothermia has limited improvement (32℃-4hr). However, the level of inflammatory inhibitor protein IL-10 is basically stable, as shown in the results. Figure 2 ; It shows that the white blood cells are in a high-energy state after stress treatment.

[0039] Example 2: Preparation of the non-aluminum-based antiperspirant and body odor suppressing composition of the present invention (abbreviated as Deswodor-Lac)

[0040] 1) Accurately weigh 10 g of chitosan powder (Sigma, CAS 9012-76-4) with a degree of deacetylation of 95%.

[0041] 2) Accurately weigh 35 g of chitosan oligosaccharide powder (Sigma, CAS 148411-57-8) with a degree of polymerization of 2-20 and a molecular weight of 2000;

[0042] 3) Accurately weigh 0.1 g of homemade high-energy leukocyte extract.

[0043] 4) Accurately weigh 5 g of vitamin B6 / pyridoxine (Sigma-Aldrich, CAS 65-23-6) in the dark.

[0044] 5) Accurately weigh 1 mg of curcumin (Fluka, CAS 458-37-7) in the dark.

[0045] 6) The above components were thoroughly mixed to obtain the intermediate product Deswodor (each 50.101g of powder contained 10g of chitosan, 35g of chitosan oligosaccharide, 0.1g of high-energy leukocyte extract, 5g of vitamin B6, and 1mg of curcumin).

[0046] 7) Accurately weigh 5 g of lyophilized Lactobacillus bulgaricus powder (prebiotic peptide source, ATCC11842) with a concentration of 10 billion CFU / g.

[0047] 8) The probiotic powder was mixed with 50.101 g of the intermediate powder to obtain the final product Deswodor-Lac.

[0048] Example 3: Determination of the microbial inhibitory function of Deswodor

[0049] 1) Dissolution of the intermediate product Deswodor: 50 g of Deswodor was fully dissolved in 500 ml of 1xPBS (BioLab), and the insoluble matter was removed by centrifugation at 3000 rpm. The supernatant was sterile filtered through a 0.22 μm filter membrane.

[0050] 2) Microbial Culture: Escherichia coli and Staphtlococcus aureus were cultured in LB liquid medium (Solarbio) at 37°C with a shaker at 250 rpm until the logarithmic growth phase. Lactobacillus bulgaricus was cultured in MRS broth (Solarbio) at 44°C with a shaker at 250 rpm until the logarithmic growth phase.

[0051] 3) Colony plate preparation: Prepare colony plates by thoroughly mixing 500 μl of each of Escherichia coli and Staphylococcus aureus (colony count 107-108 / ml) with 15 ml of 42°C solid LB medium (Solarbio). Prepare colony plates by thoroughly mixing 500 μl of Lactobacillus bulgaricus (colony count 107-108 / ml) with 15 ml of 48°C MRS solid medium (Solarbio).

[0052] 4) Use a 3 mm sterile punch to punch holes in the colony plate agar; add 300 μl of fully dissolved antiperspirant powder solution to the holes.

[0053] 5) Place the plate upright in the refrigerator for 3 hours to allow the sample to fully diffuse into the agar layer. Incubate the plate upside down for 24 hours at 37°C (for Escherichia coli and Staphylococcus aureus) and 44°C (for Lactobacillus). Remove the plate and measure the diameter of the inhibition zone. Repeat at least 3 times for each sample and take the average value. Figure 3 The intermediate product Deswodor has a significant inhibitory effect on representative microorganisms related to axillary odor (Escherichia coli and Staphylococcus aureus), and the inhibitory effect is somewhat lower than that of 75% ethanol; and this inhibition shows obvious concentration dependence. As the dilution multiple increases, the inhibitory effect is significantly weakened, showing the obvious working physicochemical characteristics of biological antibacterial preparations; but the inhibitory effect on lactobacilli is weaker, indicating that Deswodor selectively inhibits different bacterial groups, and the probiotic colonization of Deswodo-Lac in the later stage will not be affected.

[0054] Example 4: Effect of Deswodor on sweat gland cells

[0055] 1) Preparation of sweat gland cell culture medium: DMEM / F12 medium containing 5% fetal bovine serum, 10 μg / L GF, 2 μmol / L triiodothyronine, 0.4 mg / L hemisuccinyl hydrocortisone, 10 mg / L insulin-transferrin-sodium selenite, 1x 10 5 u / L penicillin and 0.1g / L streptomycin sulfate.

[0056] 2) Sweat gland cell culture: Primary sweat gland cells in a stable state (normal human apocrine sweat gland cells and apocrine sweat gland cells collected from the armpits of patients with bromhidrosis) were cultured at a cell density of 2×10 4 Cells were inoculated into 5 ml of sweat gland cell culture medium and cultured under the conditions of 5% CO2 and saturated humidity of 95%. The medium was changed half every 2 to 3 days for 8 consecutive days. The cells were examined under a microscope every day, and the cell count and viability were calculated. The results are shown in the table. Figure 4 .

[0057] 3) Effect of the intermediate product Deswodor on the growth of sweat gland cells: 1 ml of the Deswodor solution in Example 3 was added to the sweat gland cell culture medium, and the effect of Deswodor on the growth of sweat gland cells was determined according to the method in step 2; the sweat gland cells showed a typical cell proliferation curve, but the plateau phase was significantly longer; the addition of Deswodor did not significantly inhibit the growth of cells, as shown in the results. Figure 4The culture of sweat gland cells is more difficult, showing an increase in the percentage of dead cells stained during cell culture, but Deswodor treatment did not significantly change cell viability. Figure 5 .

[0058] 4) Effects on biological characteristics of deswodor sweat gland cells: RT-PCR kits were used to compare the transcriptional changes of inflammatory-related indicators (IL-1β, IL-2, IL-6, IL-8), inflammatory-suppressive indicators (IL-10), and axillary odor precursor transport-related indicators (ApoD, ASOB), with GAPDH as the internal reference; the results are shown in Figure 6 and Figure 7 Among them, compared with normal samples, the inflammatory indicators (IL-1β, IL-2, IL-6, IL-8) and precursor substance transport related indicators of axillary odor samples were increased, while the inflammatory inhibition indicators were basically the same. The results are shown in Figure 6 After treatment with Deswodor, the inflammatory indexes and precursor substance transport indexes of the axillary odor samples decreased significantly, but did not reach the level of normal samples. The inflammatory inhibition index was basically unaffected by the treatment method. The results are shown in Figure 7 .

[0059] Example 5: Preparation of the non-aluminum-based antiperspirant and body odor suppressing composition of the present invention (abbreviated as Deswodor-Lac)

[0060] 1) Accurately weigh 5 g of chitosan powder (Sigma, CAS 9012-76-4) with a degree of deacetylation of 95%.

[0061] 2) Accurately weigh 20 g of dry chitosan oligosaccharide powder (Sigma, CAS 148411-57-8) with a degree of polymerization of 2-20 and a molecular weight of 2000;

[0062] 3) Accurately weigh 0.5 g of homemade high-energy leukocyte extract.

[0063] 4) Accurately weigh 2 g of vitamin B6 / pyridoxine (Sigma-Aldrich, CAS 65-23-6) in the dark.

[0064] 5) Accurately weigh 0.5 mg of curcumin (Fluka, CAS 458-37-7) in the dark.

[0065] 6) The above components were thoroughly mixed to obtain the intermediate product Deswodor (each 27.5005g powder contained 5g chitosan, 20g chitosan oligosaccharide, 0.5g high-energy leukocyte extract, 2g vitamin B6, and 0.5mg curcumin).

[0066] 7) Accurately weigh 2 g of lyophilized Lactobacillus bulgaricus powder (prebiotic peptide source, ATCC11842) with a concentration of 10 billion CFU / g.

[0067] 8) The probiotic powder was mixed with 27.505 g of the intermediate powder to obtain the final product Deswodor-Lac.

[0068] Example 6: Preparation of the non-aluminum-based antiperspirant and body odor suppressing composition of the present invention (abbreviated as Deswodor-Lac)

[0069] 1) Accurately weigh 20 g of chitosan powder (Sigma, CAS 9012-76-4) with a degree of deacetylation of 95%.

[0070] 2) Accurately weigh 40 g of chitosan oligosaccharide powder (Sigma, CAS 148411-57-8) with a degree of polymerization of 2-20 and a molecular weight of 2000;

[0071] 3) Accurately weigh 0.05 g of homemade high-energy leukocyte extract.

[0072] 4) Accurately weigh 8 g of vitamin B6 / pyridoxine (Sigma-Aldrich, CAS 65-23-6) in the dark.

[0073] 5) Accurately weigh 1.5 mg of curcumin (Fluka, CAS 458-37-7) in the dark.

[0074] 6) The above components were thoroughly mixed to obtain the intermediate product Deswodor (each 68.0515g powder contained 20g chitosan, 40g chitosan oligosaccharide, 0.05g high-energy leukocyte extract, 8g vitamin B6, and 1.5mg curcumin.

[0075] 7) Accurately weigh 8 g of lyophilized Lactobacillus bulgaricus powder (prebiotic peptide source, ATCC11842) with a concentration of 10 billion CFU / g.

[0076] 8) The probiotic powder was mixed with 68.0515 g of the intermediate powder to obtain the final product Deswodor-Lac.

[0077] With reference to Examples 3 and 4, the microbial inhibition function and the effect on sweat gland cells of Deswodor prepared in Examples 5 and 6 were tested respectively, and similar results to those in Examples 3 and 4 were obtained.

[0078] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

Claims

1. A non-aluminum-based antiperspirant and body odor suppressing composition, characterized in that: The composition comprises the following components in parts by weight: 5-20 parts of chitosan dry powder, 20-40 parts of chitosan oligosaccharide dry powder, 0.05-0.5 parts of high-energy leukocyte extract freeze-dried powder, 2-8 parts of vitamin B6, 0.0005-0.0015 parts of curcumin, and 2-8 parts of probiotic freeze-dried powder.

2. The non-aluminum-based antiperspirant and body odor-inhibiting composition according to claim 1, characterized in that: The composition comprises the following components in parts by weight: 10 parts of chitosan dry powder, 35 parts of chitosan oligosaccharide dry powder, 0.1 parts of high-energy leukocyte extract freeze-dried powder, 5 parts of vitamin B6, 0.001 parts of curcumin, and 5 parts of probiotic freeze-dried powder.

3. The non-aluminum-based antiperspirant and body odor suppressing composition according to claim 1, characterized in that: The high-energy leukocyte extract is obtained through three steps: the first step is to separate the leukocytes in the blood of slaughtered animals; the second step is to remove the stress; and the third step is to crush and extract the contents.

4. The non-aluminum-based antiperspirant and body odor suppressing composition according to claim 3, characterized in that: The second step of destress treatment uses a destress treatment solution, which is composed of 0.9% NaCl, 1% hydroxyethyl starch, 0.125% taurine, 0.325% sodium ascorbyl phosphate, 1.5% ectoine, 1% proline, and 2% sodium butyrate in mass percentage. The destressing is completed by incubating at 32°C, 5% CO2 for 4-6 hours at a volume ratio of the treatment solution to the white blood cell suspension of 10:

1.

5. The non-aluminum-based antiperspirant and body odor suppressing composition according to claim 3, characterized in that: The slaughtered animals are rabbits, horses, cattle, sheep, deer and pigs.

6. The non-aluminum-based antiperspirant and body odor-inhibiting composition according to claim 1, characterized in that: The probiotic freeze-dried powder is one or more of Lactobacillus bulgaricus freeze-dried powder, Lactobacillus delbrueckii family freeze-dried powder, probiotic yeast freeze-dried powder, and Bifidobacterium freeze-dried powder.

7. The non-aluminum-based antiperspirant and body odor-inhibiting composition according to claim 6, characterized in that: The concentration of the lyophilized Lactobacillus bulgaricus powder is 10 billion CFU / g.

8. The non-aluminum-based antiperspirant and body odor suppressing composition according to claim 1, characterized in that: It is characterized by: The composition is prepared as dry powder, aqueous solution, emulsion or colloid.

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