A composition for protecting joint sub-health and external preparation thereof

Abstract

The application discloses a composition for protecting joint sub-health and an external preparation thereof, and belongs to the technical field of biological preparations. The technical problem to be solved is to provide a composition which can achieve the balance between the treatment and prevention by synergistically regulating cartilage inflammation, cartilage matrix metabolism and osteogenic differentiation and mineralization process through multiple targets and multiple pathways. The composition comprises mousse cream and radix rhei extract liquid with a mass ratio of 10-40:1.

Description

Technical Field

[0001] This invention belongs to the field of biopharmaceutical technology, specifically relating to a composition for protecting joint sub-health and its topical formulation. Background Technology

[0002] Joint sub-health refers to an intermediate state between health and disease in the joints and surrounding tissues. It often manifests as mild joint soreness, morning stiffness, and discomfort after activity. Without intervention, it may further develop into chronic diseases such as osteoarthritis and rheumatoid arthritis, significantly negatively impacting motor function and quality of life. With the accelerating aging population and the increasing number of people suffering from sports injuries, joint sub-health is becoming increasingly prevalent, leading to a strong market demand for safe and effective topical joint care products.

[0003] The metabolic balance of articular cartilage depends on the dynamic regulation between anabolism (synthesis of COL-II and proteoglycans) and catabolism (degradation of ECM by matrix metalloproteinases such as MMPs and ADAMTS) within chondrocytes. Under normal conditions, the two metabolic processes maintain a balance; however, when subjected to mechanical injury, age-related degeneration, or continuous stimulation by inflammatory factors (such as IL-1β, IL-6, and TNF-α), catabolism becomes dominant, leading to progressive loss of cartilage matrix. IL-1β is a core pro-inflammatory cytokine mediating cartilage catabolism, significantly inhibiting the gene expression of COL-II and Aggrecan, and upregulating the activity of matrix-degrading enzymes such as MMP-13 and ADAMTS-5, thereby accelerating cartilage destruction. IL-6 mainly participates in synovial inflammation and is closely related to joint swelling and pain. Therefore, protecting articular cartilage, inhibiting the secretion of inflammatory factors, and promoting COL-II synthesis are core strategies for intervening in sub-optimal joint health.

[0004] Osteoblasts, derived from the directed differentiation of bone marrow mesenchymal stem cells, play a crucial role in bone formation and metabolism. During osteophyte formation and bone repair at the joint-bone interface, the differentiation and mineralization activity of osteoblasts directly affect bone health. Studies have shown that moderately promoting osteoblast differentiation helps maintain the integrity of periarticular bone tissue and prevents osteoporosis-related joint instability. Alkaline phosphatase (ALP) is a marker enzyme for early osteoblast differentiation; its activity directly reflects the osteogenic induction effect. Alizarin red staining is used to assess the degree of osteoblast mineralization nodule formation and is the gold standard indicator of osteoblast functional maturity.

[0005] Joint sub-health is an intermediate state between health and organic joint disease. Current clinical interventions primarily focus on symptomatic treatment. Various conventional treatment plans have significant limitations and potential risks, making them unsuitable for the long-term daily maintenance needs of the population. Specifically: (1) Oral anti-inflammatory and analgesic drugs: Clinically commonly used non-steroidal oral drugs (celecoxib, ibuprofen, etc.) can quickly suppress pain and relieve acute inflammation, but they can only treat the symptoms and cannot reverse the loss of cartilage matrix or regulate the process of osteogenic differentiation. Long-term continuous use can easily induce gastrointestinal ulcer bleeding, liver and kidney damage, and increase the risk of adverse cardiovascular and cerebrovascular events. The use of drugs is severely limited for the elderly and people with chronic diseases. It is only recommended for acute phase and short-term intermittent use. It is not suitable for long-term preventive maintenance of joint sub-healthy people.

[0006] (2) Intra-articular injection therapy: Joint injections such as glucocorticoids have a rapid onset of action and can quickly eliminate joint swelling and pain. However, repeated injections of glucocorticoids will directly accelerate chondrocyte apoptosis and aggravate matrix degradation. Moreover, the injection is an invasive procedure with risks of joint infection, local swelling and pain, and tendon damage. It needs to be performed regularly at medical institutions, which has a high operation threshold and cannot be used as a daily care method.

[0007] (3) Surgical repair / replacement therapy: Unicompartmental osteotomy and artificial joint replacement are only suitable for mid-to-late stage severe osteoarthritis. They are end-stage salvage options, which are highly invasive, have a long postoperative recovery period, and have complications such as prosthesis wear and loosening, postoperative infection, and nerve stimulation. For sub-healthy joint patients who only have soreness and morning stiffness, there are no indications for surgery. The intervention cost is extremely high and it does not have early protective value.

[0008] (4) Hospital physical therapy and equipment assistance: Infrared, shock wave, ultrasound, hot compress therapy device, joint brace, etc. can only improve local blood circulation and relax the surface soft tissue, but cannot act on the deep cartilage metabolic pathway; shock wave therapy has the risk of subcutaneous hematoma, skin burn and nerve stimulation, and long-term use of heat lamp can easily cause skin pigmentation; moreover, physical therapy requires going to medical institutions at fixed times, and home devices are bulky and inconvenient to carry, making it difficult to integrate into fragmented daily scenarios such as commuting, sports and home, and can only be used as an intermittent auxiliary relief method, and cannot continuously and effectively maintain joints.

[0009] Patent document CN113332405A (publication date September 3, 2021) discloses a biological agent for preventing and alleviating joint sub-health. The biological agent includes components such as sodium chondroitin sulfate, extract of *Lycopodium clavatum*, extract of *Luffa cylindrica*, extract of ginger root, extract of *Morinda officinalis*, frankincense oil, methyl salicylate, menthol, and camphor. This invention is administered through the skin in the form of a liquid preparation, which can effectively prevent and alleviate joint inflammation.

[0010] Patent document CN119818625A (publication date April 15, 2025) discloses a topical composition for preventing and relieving joint discomfort. This topical composition includes: Panax notoginseng extract, Uncaria rhynchophylla extract, Geranium wilfordii extract, Carthamus tinctorius extract, Clematis chinensis extract, Angelica sinensis root extract, Aconitum carmichaelii extract, Saussurea involucrata extract, Pheretima aspergillum extract, Corydalis yanhusuo extract, Sparganium stoloniferum extract, Achyranthes bidentata extract, Cinnamomum cassia extract, Morinda officinalis extract, Pinus massoniana extract, Lycopodium clavatum extract, Liquidambar formosana extract, sodium chondroitin sulfate, dimethyl sulfone, Piper nigrum extract, and Zingiber officinale root extract. This invention, through the meridian-tropism of each component, induces the active ingredients to directly reach the site of joint discomfort; it can quickly relieve symptoms such as numbness and pain, regulate function, and promote recovery.

[0011] Existing topical joint preparations have advantages such as ease of use, integration into daily life, and low risk, forming a new approach to joint care. However, they still have the following problems: (1) poor permeability of active ingredients and short duration of efficacy; (2) single formulation, making it difficult to achieve comprehensive intervention on multiple targets; (3) lack of verification of the synergistic mechanism of osteoblast differentiation and chondrocyte protection. Therefore, developing a composition with synergistic effects to achieve multi-target and multi-mechanism combined intervention for sub-healthy joints has significant technological innovation value and market application prospects. Summary of the Invention

[0012] The purpose of this invention is to provide a composition for protecting against sub-health of joints and related technologies, in order to solve technical problems such as improving the efficacy of protecting against sub-health of joints, or a combination thereof.

[0013] Terminology Explanation: Unless otherwise defined, all technical terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms, the definition provided in this chapter shall prevail.

[0014] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0015] Definitions of standard chemical terms can be found in the textbook or reference book "Pharmacopoeia of the People's Republic of China: 2025 Edition".

[0016] Unless otherwise specified, conventional methods within the scope of the art, such as cell proliferation activity assay, alkaline phosphatase activity assay, extracellular matrix mineralization staining, cartilage tissue pathological staining, inflammatory factor ELISA quantification, real-time quantitative PCR, in vitro cartilage tissue culture evaluation, animal joint modeling and behavior, tissue HE pathological staining, and ultrasound imaging, shall be used.

[0017] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0018] Terminology Explanation: Joint sub-health refers to an intermediate sub-healthy state between health and organic lesions in the joints and surrounding soft tissues. It is characterized by joint soreness, morning stiffness, discomfort during movement, difficulty in flexion and extension, mild synovitis and cartilage metabolic imbalance. It is a pre-pathological state that has not yet developed into a clear joint disease such as osteoarthritis or rheumatoid arthritis.

[0019] Composition: This invention specifically refers to a topical efficacy complex system composed of a specific mass ratio of mousse cream and Lulutong essence, prepared according to a defined formula and preparation process, used to protect and intervene in sub-healthy joints.

[0020] Topical formulations: Various topical dosage forms comprising the compositions described in this invention, supplemented with excipients acceptable in biological agents, which can be individually packaged or packaged in sets, for use in the preparation of topical drugs / skin care products / medical devices to protect against joint sub-health.

[0021] Synergistic effect: This refers to the overall efficacy of the mousse cream and Lulutong essence in anti-inflammatory, osteoblast differentiation promotion, extracellular matrix mineralization promotion, type II collagen and glycosaminoglycan synthesis, etc., which is significantly higher than the sum of the effects of the two drugs alone. This indicates that the combined administration is more effective than the single administration and that the two drugs have a synergistic effect.

[0022] Human osteoblasts are functional cells derived from human bone marrow mesenchymal stem cells, possessing directed differentiation and mineralization capabilities. They participate in periarticular bone metabolism, bone repair, and bone homeostasis maintenance, and are a classic in vitro cell model for evaluating osteogenic differentiation and mineralization capacity.

[0023] Human chondrocytes are the core functional cells of articular cartilage tissue. They are responsible for synthesizing cartilage matrix such as type II collagen and glycosaminoglycans, maintaining the integrity of cartilage structure and metabolic balance. They are prone to apoptosis and matrix degradation under inflammatory stimulation.

[0024] Extracellular matrix mineralization: The process of forming mineralized nodules in the late stage of osteoblast differentiation and maturation is a core indicator for evaluating osteogenic activity and bone formation capacity. Alizarin Red S staining is commonly used for qualitative and semi-quantitative detection.

[0025] Alkaline phosphatase (ALP): A marker enzyme for early osteoblast differentiation. Higher ALP activity indicates stronger osteoblast differentiation and osteogenic activity.

[0026] Type II collagen (COL-II): The most important structural collagen in articular cartilage, maintaining cartilage tension and integrity. Its synthesis decreases significantly under cartilage damage and inflammation.

[0027] Inflammatory factors: IL-6: Interleukin-6, a key pro-inflammatory factor, mediates inflammation, swelling and discomfort of the synovial membrane of the joint, and is a core indicator for evaluating the level of joint inflammation; IL-1β: Interleukin-1β, a core pro-inflammatory factor in cartilage catabolism, can inhibit collagen synthesis, upregulate matrix degradation enzymes, and accelerate cartilage degeneration; COX-II: Cyclooxygenase 2, a key enzyme that induces inflammation and mediates inflammatory cascades and pain responses.

[0028] LPS: Lipopolysaccharide, commonly used as an in vitro inflammation modeling agent, can induce significant inflammatory responses in chondrocytes and construct cellular inflammatory injury models.

[0029] RT-qPCR: Real-time quantitative polymerase chain reaction, used to detect the relative expression levels of mRNAs of genes related to inflammation and cartilage metabolism.

[0030] ELISA: Enzyme-linked immunosorbent assay, a routine method for quantitatively detecting the content of inflammatory factors, neuropeptides, and matrix proteins in cell / tissue supernatants.

[0031] The term "homogenization" as used in this article refers to the physical process of refining, crushing, uniformly dispersing, and emulsifying a multiphase mixture of materials, including aqueous phase, oil phase, plant extracts, powder particles, and various functional excipients, within a formulation system. This process reduces the particle size of the materials, eliminates phase stratification, inhibits precipitation, and improves the system's uniformity, fineness, and storage stability, while ensuring the uniform distribution of active ingredients. High-speed shear homogenization, high-pressure homogenization, and stirring emulsification homogenization are all conventional homogenization methods in this field.

[0032] To solve the above-mentioned technical problems, the present invention adopts the following technical solution: In a first aspect, the present invention provides: a composition for protecting joint sub-health, comprising a mousse cream and a lulutong essence in a mass ratio of 10-40:1.

[0033] In some embodiments, the mass ratio of the mousse cream to the Lulutong essence can be selected as 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1, 31:1, 32:1, 33:1, 34:1, 35:1, 36:1, 37:1, 38:1, 39:1, 40:1, or a range derived therefrom; more preferably 10-20:1.

[0034] In some embodiments, the mousse cream comprises, by weight percentage: Phase A: Water, propylene glycol 1%-5%, butylene glycol 1%-3%, disodium EDTA 0.01%-0.1%, acetylglucosamine 0.1%-1.0%, sodium chondroitin sulfate 0.01%-0.5%, lycopodium extract 0.01%-0.5%, and Luffa cylindrica extract 0.01%-0.5%; Phase B: Dipropylene glycol 1.0%-10.0%, sodium polyacrylate grafted starch 0.1%-1.0%; Phase C: Rice bran oil 1.0%-10.0%, Caprylic / Capric triglycerides 1.0%-10.0%, Isopropyl palmitate 1.0%-5.0%, Hydrogenated polyisobutylene 1.0%-4.0%, Isocerocetyl ether-20 0.1%-1%, Methyl glucetyl ether-20 0.1%-1%, Glyceryl stearate 0.1%-1%, Polydimethylsiloxane 0.5%-2.0%, Cyclopentasiloxane 0.1%-0.5%, Cyclohexylsiloxane 0.1%-0.5%, Sorbitan oleate 0.1%-0.5%, Cetearyl alcohol 0.1%-0.5%, Potassium azeloyl diglycinate 0.1%-0.5%; Phase D: Tocopherol (Vitamin E) 0.05%-0.1%, Sunflower seed oil 0.05%-0.1%; Phase E: Tetrahydropiperine 0.01%-0.1%, Propylene glycol 0.05%-1.0%; Phase F: Phenoxyethanol 0.1%-1%, Ethylhexylglycerin 0.1%-1%, Methylparaben 0.05%-0.1%, Ethylparaben 0.05%-0.1%; Phase G: Gentiana macrophylla 0.1%-2%; H phase: 0.01%-0.1% white chrysanthemum flower oil, 0.01%-0.1% rose flower oil, 0.01%-0.1% cinnamon leaf oil, 0.01%-0.14% ylang-ylang flower oil, 0.001%-0.1% ginger root oil, 0.01%-0.2% vanillyl butyl ether, 0.01%-0.2% kava pepper, 0.01%-0.3% white beech leaf oil; Remaining water.

[0035] In other embodiments, the mousse cream comprises, by weight percentage: Phase A: Water, propylene glycol 2.0-4.0%, butylene glycol 1.5-2.5%, disodium EDTA 0.03-0.07%, acetylglucosamine 0.2%-0.8%, sodium chondroitin sulfate 0.15%-0.35%, lycopodium extract 0.15%-0.35%, and Luffa cylindrica extract 0.1%-0.3%; Phase B: Dipropylene glycol 3.0%-7.0%, sodium polyacrylate grafted starch 0.15%-0.5%; Phase C: Rice bran oil 3.0%-9.0%, caprylic / capric triglycerides 5%-10%, isopropyl palmitate 2.0%-4.0%, hydrogenated polyisobutylene 1.0%-3.0%, isocetyl alcohol polyether-20 0.3%-0.7%, methyl glucetol polyether-20 0.1%-0.5%, glyceryl stearate 0.4%-0.8%, polydimethylsiloxane 0.5%-1.5%, cyclopentamethoxydimethylsiloxane 0.2%-0.4%, cyclohexylsiloxane 0.2%-0.4%, sorbitan oleate 0.2%-0.4%, cetearyl alcohol 0.2%-0.4%, potassium azeloyl diglycinate 0.2%-0.4%; Phase D: Tocopherol 0.06%-0.1%, Sunflower seed oil 0.06%-0.1%; Phase E: Tetrahydropiperine 0.06%-0.1%, Propylene glycol 0.06%-0.1%; Phase F: Phenoxyethanol 0.3%-0.5%, Ethylhexylglycerin 0.03%-0.5%, Methylparaben 0.05%-0.08%, Ethylparaben 0.05%-0.08%; Phase G: Gentiana macrophylla 0.5%-1.5%; H phase: 0.03%-0.07% white chrysanthemum flower oil, 0.03%-0.07% rose flower oil, 0.03%-0.07% cinnamon leaf oil, 0.03%-0.10% ylang-ylang flower oil, 0.06%-0.09% ginger root oil, 0.05%-0.12% vanillyl butyl ether, 0.4%-1.4% kava pepper, 0.15%-0.25% white beech leaf oil; Remaining water.

[0036] In other embodiments, the mousse cream comprises, by weight percentage: Phase A: Water, propylene glycol 3.0%, butylene glycol 2.0%, disodium EDTA 0.05%, acetylglucosamine 0.5%, sodium chondroitin sulfate 0.25%, lycopodium extract 0.25%, and Luffa cylindrica extract 0.2%; Phase B: Dipropylene glycol 5.0%, sodium polyacrylate grafted starch 0.2%; Phase C: Rice bran oil 6.0%, Caprylic / Capric triglyceride 8.0%, Isopropyl palmitate 3.0%, Hydrogenated polyisobutylene 2.0%, Isocerocetyl ether-20 0.5%, Methyl glucetyl ether-20 0.3%, Glyceryl stearate 0.6%, Polydimethylsiloxane 1.0%, Cyclopentasiloxane 0.3%, Cyclohexylsiloxane 0.3%, Sorbitan oleate 0.3%, Cetearyl alcohol 0.3%, Potassium azeloyl diglycinate 0.3%; Phase D: Tocopherol (Vitamin E) 0.08%, Sunflower Seed Oil 0.08%; Phase E: Tetrahydropiperine 0.08%, Propylene glycol 0.08%; Phase F: Phenoxyethanol 0.5%, Ethylhexylglycerin 0.5%, Methylparaben 0.08%, Ethylparaben 0.08%; G phase: Gentiana macrophylla 1.0%; H phase: 0.05% white chamomile flower oil, 0.05% rose flower oil, 0.05% cinnamon leaf oil, 0.07% ylang-ylang flower oil, 0.08% ginger root oil, 0.08% vanillyl butyl ether, 0.9% kava pepper, 0.2% flat-leaved white beech leaf oil; Remaining water.

[0037] In some embodiments, the method for preparing the mousse cream includes the following steps: Step 1: After stirring and dissolving phase A and homogenizing it, keep it warm for later use; Step 2: Cool down, add phase C and phase D, and stir well; Step 3: Continue adding phase B and homogenizing, then add phase E and mix thoroughly; Step 4: Cool down to below 45℃, add phase F, phase G and phase H, stir well, and it's ready.

[0038] Preferably, the insulation temperature in step 1 is 92-95℃, such as 92℃, 93℃, 94℃, 95℃, or a range derived therefrom.

[0039] Preferably, the cooling temperature in step 2 is 85-88°C, such as 85°C, 86°C, 87°C, 88°C, or a range derived therefrom.

[0040] In some embodiments, the Lulutong essence comprises, by weight percentage: Phase A: Water, propylene glycol 1%-15%, magnesium aluminum silicate 0.05%-0.3%, sodium chondroitin sulfate 0.01%-0.5%, lycopodium extract 0.01%-0.2%, and Luffa cylindrica extract 0.01%-0.2%; Phase B: Sodium stearoyl glutamate 0.10%-0.2%, lauryl ether-3 0.5%-3%, sorbitan sesquioleate 0.1%-1%, sorbitan trioleate 0.1%-1%, sorbitan stearate 0.1%-0.5%; Phase C: 0.1%-1% acrylic (ester) copolymers, 0.1%-0.5% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 0.10%-0.5% isohexadecane, 0.01%-0.5% polysorbate-60, 0.01%-0.5% sorbitan isostearate; Phase D: Frankincense (BOSWELLIA CARTERII) oil 0.1%-1%, methyl salicylate 0.1%-1%, Ceylon cinnamon leaf oil 0.1%-1%, lemongrass leaf oil / citronella oil 0.1%-1%, vanillyl butyl ether 0.05%-1.5%, camphor 0.1%-4%, menthol 0.1%-1.5%, butylated hydroxytoluene 0.01%-0.2%, tetrahydropiperine 0.01%-0.2%; Phase E: Triethanolamine 0.05%-0.2%, Citric Acid 0.01%-0.2%; Phase F: 1,2-Pentanediol 0.10%-0.5%, Butylene glycol 0.1%-2%, Hydroxyphenylpropionamide benzoic acid 0.01%-0.5%, Ascorbyl palmitate 0.1%-0.5%, Glycerin 0.1%-5%, Ginger root extract 0.01%-1%, Potassium sorbate 0.01%-0.5%, Sodium benzoate 0.01%-0.5%, Morinda officinalis fruit extract 0.01%-0.5%; Phase G: Phenoxyethanol 0.1%-1%, Ethylhexylglycerin 0.1%-0.5%, Methylparaben 0.01%-0.1%, Ethylparaben 0.01%-0.1%; Remaining water.

[0041] In other embodiments, the Lulutong essence comprises, by weight percentage: Phase A: Water, propylene glycol 8.0%-12.0%, magnesium aluminum silicate 0.05%-0.15%, sodium chondroitin sulfate 0.2%-0.4%, lycopodium extract 0.05%-0.15%, and Luffa cylindrica extract 0.05%-0.15%; Phase B: Sodium stearoyl glutamate 0.12%-0.18%, lauryl ether-3 1.5%-2.5%, sorbitan sesquioleate 0.3%-0.7%, sorbitan trioleate 0.3%-0.7%, sorbitan stearate 0.2%-0.4%; Phase C: 0.5%-0.9% acrylic (ester) copolymers, 0.2%-0.4% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 0.2%-0.4% isohexadecane, 0.25%-0.35% polysorbate-60, and 0.2%-0.4% sorbitan isostearate. Phase D: Frankincense (BOSWELLIA CARTERII) oil 0.2%-0.8%, methyl salicylate 0.2%-0.8%, Ceylon cinnamon leaf oil 0.2%-0.8%, lemongrass leaf oil / citronella oil 0.2%-0.8%, vanillyl butyl ether 0.07%-1.2%, camphor 1%-3%, menthol 0.8%-1.2%, butylated hydroxytoluene 0.05%-0.15%, tetrahydropiperine 0.05%-0.15%; Phase E: Triethanolamine 0.05%-0.15%, Citric Acid 0.05%-0.15%; Phase F: 1,2-Pentanediol 0.10%-0.3%, Butylene glycol 0.5%-2%, Hydroxyphenylpropionamide benzoic acid 0.1%-0.3%, Ascorbyl palmitate 0.2%-0.4%, Glycerin 0.2%-0.5%, Ginger root extract 0.2%-0.8%, Potassium sorbate 0.1%-0.3%, Sodium benzoate 0.1%-0.3%, Morinda officinalis fruit extract 0.2%-0.4%; Phase G: Phenoxyethanol 0.2%-0.5%, Ethylhexylglycerin 0.1%-0.3%, Methylparaben 0.02%-0.05%, Ethylparaben 0.02%-0.05%; Remaining water.

[0042] In other embodiments, the Lulutong essence comprises, by weight percentage: Phase A: Water, 10% propylene glycol, 0.1% magnesium aluminum silicate, 0.3% sodium chondroitin sulfate, 0.1% lycopodium extract, 0.1% luffa extract; Phase B: Sodium stearoyl glutamate 0.15%, lauryl ether-3 2%, sorbitan sesquioleate 0.5%, sorbitan trioleate 0.5%, sorbitan stearate 0.3%; Phase C: 0.7% acrylic (ester) copolymer, 0.3% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 0.3% isohexadecane, 0.3% polysorbate-60, 0.3% sorbitan isostearate; Phase D: 0.5% Boswellia Carterii oil, 0.5% methyl salicylate, 0.5% Ceylon cinnamon leaf oil, 0.5% lemongrass leaf oil / citronella oil, 1% vanillyl butyl ether, 2% camphor, 1% menthol, 0.1% butylated hydroxytoluene, 0.1% piperine; Phase E: Triethanolamine 0.1%, Citric Acid 0.1%; Phase F: 1,2-Pentanediol 0.2%, Butylene glycol 1%, Hydroxyphenylpropionamide benzoic acid 0.2%, Ascorbyl palmitate 0.3%, Glycerin 0.3%, Ginger root extract 0.5%, Potassium sorbate 0.2%, Sodium benzoate 0.3%, Morinda officinalis fruit extract 0.3%; Phase G: Phenoxyethanol 0.5%, Ethylhexylglycerin 0.3%, Methylparaben 0.05%, Ethylparaben 0.05%; Remaining water.

[0043] In some embodiments, the preparation method of the Lulutong essence includes the following steps: Step 1: After stirring and dissolving phase A and homogenizing it, keep it warm for later use; Step 2: Cool down, add phase B, and stir well; Step 3: Continue adding phase D and homogenizing, then add phase C and homogenize. Step 4: Cool down to below 45℃, add phase E, phase F and phase G, stir well, and it's ready.

[0044] Preferably, the insulation temperature in step 1 is 92-95℃, such as 92℃, 93℃, 94℃, 95℃, or a range derived therefrom.

[0045] Preferably, the cooling temperature in step 2 is 85-88°C, such as 85°C, 86°C, 87°C, 88°C, or a range derived therefrom.

[0046] In a second aspect, the present invention provides: a topical preparation for protecting against joint sub-health, comprising the aforementioned composition.

[0047] The protection described in this invention includes at least one of the following: relieving joint soreness and swelling, improving joint function, inhibiting the expression of chondrocyte inflammatory factor IL-6, promoting collagen COL-II synthesis, promoting osteoblast differentiation, and promoting osteoblast mineralization.

[0048] In some embodiments, the composition can be used alone as a topical preparation to protect against joint sub-health.

[0049] In some implementations, the mousse cream and Lulutong essence in the topical preparation can be packaged separately or combined into a set of products for unified packaging.

[0050] In some embodiments, the composition can be used as an active ingredient in the preparation of topical formulations.

[0051] In some embodiments, the topical formulation may also include acceptable excipients.

[0052] In some embodiments, the topical preparation may include ointments, creams, gels, liniments, coatings, films, tinctures, lotions, powders, patches, plasters, films, sprays, or aerosols.

[0053] In some embodiments, the excipients include at least one of pressure-sensitive adhesives, foaming agents, defoamers, adsorbents, absorbents, colorants, fragrances, pH adjusters, osmotic pressure adjusters, buffers, antibacterial agents, antioxidants, antioxidant synergists, chelating agents, preservatives, humectants, skin penetration enhancers, softeners, thickeners, plasticizers, encapsulating agents, suspending agents, film-forming agents, anti-adhesion agents, flow aids, anti-caking agents, emulsifiers, solubilizers, cosolvents, surfactants, solvents, propellants, ointment bases, and carrier materials.

[0054] In some embodiments, the preparation method of the topical formulation is selected from the methods described in the general rules for preparation of the Chinese Pharmacopoeia, the Japanese Pharmacopoeia, the United States Pharmacopoeia, and the European Pharmacopoeia. The specific preparation method depends on the dosage form.

[0055] Based on further solutions to the technical problems of the present invention, or simultaneous solutions to multiple technical problems, the preferred solution in the technical solution provided in the first aspect of the present invention includes: The mass ratio of the mousse cream to the Lulutong essence is 10-40:1. This technical solution, while improving the efficacy of protecting against sub-healthy joints, further produces a synergistic effect, strengthening the efficacy of protecting against sub-healthy joints.

[0056] In this invention, Examples 1-3 at least support the protection scope of "the mass ratio of the mousse cream and the Lulutong essence".

[0057] The mass ratio of mousse cream to Luffa cylindrica essence is 10:1, 20:1, and 40:1 respectively, as explained above and / or in Examples 1-3, summarized by the common feature "mass ratio of mousse cream to Luffa cylindrica essence". Therefore, those skilled in the art can reasonably infer that the technical feature of a mass ratio of mousse cream to Luffa cylindrica essence of 10-40:1, its subordinate concepts, substantially equivalent technical means, and technical means that can replace the mass ratio of mousse cream to Luffa cylindrica essence of 10-40:1 based on existing technology and conventional technical means and common knowledge should all fall within the protection scope of this invention. For example, replacing the mass ratio of mousse cream to Luffa cylindrica essence with 11:1, 39:1, 19:1, 21:1, etc., while keeping other technical features unchanged, still falls within the protection scope of this invention.

[0058] The present invention has at least the following beneficial effects: 1. Compared with existing technologies, this invention provides a topical composition for protecting against joint sub-health. Utilizing the synergistic effects of the components in the mousse cream and the Lulutong essence, it exhibits significant protective and improving effects on joint sub-health. This composition achieves a comprehensive intervention effect by synergistically regulating cartilage inflammation, cartilage matrix metabolism, and osteogenic differentiation and mineralization processes through multiple targets and pathways. It is particularly suitable for joint sub-health conditions caused by joint soreness, limited range of motion, mild synovitis, and cartilage metabolic imbalance.

[0059] 2. This invention specifies a particular compounding ratio of 10-40:1 between the mousse cream and the Lulutong essence. Experimental data show that the composition within the specified ratio range of this invention exhibits a synergistic effect in promoting extracellular matrix mineralization and enhancing type II collagen synthesis, both significantly superior to single mousse cream or single Lulutong essence.

[0060] 3. This invention provides a topical composition and pharmaceutical preparation with clearly defined efficacy targets, outstanding synergistic advantages, and good safety. It is an important supplement and technological optimization to existing joint sub-health protection products and has significant clinical application value. Attached Figure Description

[0061] Figure 1 The results are for extracellular matrix mineralization detection. Compared to the NC group, #P < 0.05; compared to the MC group, P < 0.05, compared to group A; ^ P < 0.05; Figure 2 The results show the content of type II collagen and glycosaminoglycans in human chondrocytes in vitro. Compared to the NC group, #P < 0.05; compared to the MC group, P < 0.05, compared to group A; ^ P < 0.05; Figure 3 The results of behavioral change detection are shown in Figure a, which is a statistical graph of the average footprint area, and Figure b, which is a statistical graph of the hind limb stride width. Compared to the MC group, P < 0.05; P < 0.01; Figure 4 The graph shows the statistical results of relevant indicators in joint tissue. I represents the IL-6 content in joint tissue, II represents the substance P content in joint tissue, and III represents the CGRP content in joint tissue. Different letters indicate significant differences between groups (P < 0.05). Figure 5 This is a statistical graph showing the IL-6 content in skin tissue. Different letters indicate significant differences between groups (P < 0.05). Detailed Implementation

[0062] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0063] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0064] All numerical values ​​or expressions relating to component amounts, process conditions, etc., used in this invention shall be understood to be modified by the word "about" in all cases. When referring to a quantity or range of values, the quantity or range is an approximation within experimental variability (or within statistical experimental error). In this invention, the term "about" shall have the meaning of being within 10%, preferably within 5%, of the specified value or range.

[0065] Unless otherwise specified, percentages in this invention refer to mass percentages, temperatures refer to room temperature, and solvents refer to water.

[0066] In this document, “room temperature” refers to ambient temperature, ranging from about 10°C to about 40°C. In some embodiments, “room temperature” refers to a temperature ranging from about 20°C to about 30°C; in other embodiments, “room temperature” refers to a temperature ranging from about 25°C to about 30°C; and in still other embodiments, “room temperature” refers to 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, etc.

[0067] Acetyl glucosamine, purchased from HeYi Commercial Port (Guangzhou) Trade Development Co., Ltd., item number RH2024100602; sodium chondroitin sulfate, purchased from Shandong Zhongshan Biotechnology Co., Ltd., item number CSB25001; lycopodium extract, purchased from Xi'an Weite Biotechnology Co., Ltd., item number DC. SS-20250922; *Lysimachia christinae* extract, purchased from Xi'an Weite Biotechnology Co., Ltd., item number DCLLT-20251210; Sodium polyacrylate grafted starch, purchased from Guangzhou Hengtao Trading Co., Ltd., item number K-52012; Rice bran oil, purchased from Guangzhou Siyi Biotechnology Co., Ltd., item number SE20250923; Sunflower seed oil, purchased from Guangzhou Nuoran Biotechnology Co., Ltd., item number CH9000248; Gentiana macrophylla, purchased from Guangzhou Senken Biotechnology Co., Ltd., item number 2024011901; White chrysanthemum flower oil, purchased from Foshan Ruiheng Biotechnology Co., Ltd., item number 20251208; Rose flower oil, purchased from Guangzhou... The following oils were purchased from various companies: Zhouyueze Chemical Co., Ltd. (item number MFE0925051811); Cinnamon leaf oil (item number 2025.05.12, purchased from Ji'an Zhongxiang Natural Plant Co., Ltd.); Ylang-ylang oil (item number 2026.01.12, purchased from Ji'an Zhongxiang Natural Plant Co., Ltd.); Ginger root oil (item number GOL1010L10-2025102301, purchased from Guangzhou Nuoran Biotechnology Co., Ltd.); Kava pepper (item number PMM3001-260306, purchased from Xi'an Lvtian Biotechnology Co., Ltd.); White beech leaf oil (item number 2024030301, purchased from Foshan Lvcuiyuan Biotechnology Co., Ltd.); Frankincense (BOSWELLIA). Carterii oil, purchased from Guangzhou Yuxin Biotechnology Co., Ltd., product number K-2490-KE-2025; Ceylon cinnamon leaf oil, purchased from Ji'an Zhongxiang Natural Plant Co., Ltd., product number 2025.11.18; Lemongrass (Cymbopogon) CITRATUS leaf oil, purchased from Foshan Ruiheng Biotechnology Co., Ltd., item number 20250728; ginger root extract, purchased from Guangzhou Nuoran Biotechnology Co., Ltd., item number 2573340; Morinda officinalis fruit extract, purchased from Guangzhou Nuoran Biotechnology Co., Ltd., item number 2446216; hydrogenated polyisobutylene, purchased from Shanghai Xinyan International Trade Co., Ltd., item number 2601MCL003; polydimethylsiloxane, purchased from Guangzhou Guyi Chemical Technology Co., Ltd., item number H135Q2R001; tetrahydropiperine, purchased from Hainan Ruiding Chemical Trade Co., Ltd., item number HN260056; potassium azeloyl diglycinate, purchased from Guangzhou Gude Trade Co., Ltd., item number 0250000019.

[0068] Examples 1-3 Composition for protecting against joint sub-health The formulations of the compositions used in Examples 1-3 for protecting against joint sub-health are shown in Table 1.

[0069] Table 1

[0070] The mousse cream, by weight percentage, comprises: Phase A: Water, propylene glycol 3.0%, butylene glycol 2.0%, disodium EDTA 0.05%, acetylglucosamine 0.5%, sodium chondroitin sulfate 0.25%, lycopodium extract 0.25%, and Luffa cylindrica extract 0.2%; Phase B: Dipropylene glycol 5.0%, sodium polyacrylate grafted starch 0.2%; Phase C: Rice bran oil 6.0%, Caprylic / Capric triglyceride 8.0%, Isopropyl palmitate 3.0%, Hydrogenated polyisobutylene 2.0%, Isocerocetyl ether-20 0.5%, Methyl glucetyl ether-20 0.3%, Glyceryl stearate 0.6%, Polydimethylsiloxane 1.0%, Cyclopentasiloxane 0.3%, Cyclohexylsiloxane 0.3%, Sorbitan oleate 0.3%, Cetearyl alcohol 0.3%, Potassium azeloyl diglycinate 0.3%; Phase D: Tocopherol (Vitamin E) 0.08%, Sunflower Seed Oil 0.08%; Phase E: Tetrahydropiperine 0.08%, Propylene glycol 0.08%; Phase F: Phenoxyethanol 0.5%, Ethylhexylglycerin 0.5%, Methylparaben 0.08%, Ethylparaben 0.08%; G phase: Gentiana macrophylla 1.0%; H phase: 0.05% white chamomile flower oil, 0.05% rose flower oil, 0.05% cinnamon leaf oil, 0.07% ylang-ylang flower oil, 0.08% ginger root oil, 0.08% vanillyl butyl ether, 0.9% kava pepper, 0.2% flat-leaved white beech leaf oil; Remaining water.

[0071] The preparation method of the mousse cream is as follows: Step 1: After stirring and dissolving phase A and homogenizing, keep it at 92℃ for later use; Step 2: Cool down to 85℃, add phase C and phase D, and stir until homogeneous; Step 3: Continue adding phase B and homogenizing, then add phase E and mix thoroughly; Step 4: Cool down to 35℃, add phase F, phase G and phase H, stir well, and it's ready.

[0072] The Lulutong essence, by mass percentage, comprises: Phase A: Water, 10% propylene glycol, 0.1% magnesium aluminum silicate, 0.3% sodium chondroitin sulfate, 0.1% lycopodium extract, 0.1% luffa extract; Phase B: Sodium stearoyl glutamate 0.15%, lauryl ether-3 2%, sorbitan sesquioleate 0.5%, sorbitan trioleate 0.5%, sorbitan stearate 0.3%; Phase C: 0.7% acrylic (ester) copolymer, 0.3% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 0.3% isohexadecane, 0.3% polysorbate-60, 0.3% sorbitan isostearate; Phase D: 0.5% Boswellia Carterii oil, 0.5% methyl salicylate, 0.5% Ceylon cinnamon leaf oil, 0.5% lemongrass leaf oil, 1% vanillyl butyl ether, 2% camphor, 1% menthol, 0.1% butylated hydroxytoluene, 0.1% piperine; Phase E: Triethanolamine 0.1%, Citric Acid 0.1%; Phase F: 1,2-Pentanediol 0.2%, Butylene glycol 1%, Hydroxyphenylpropionamide benzoic acid 0.2%, Ascorbyl palmitate 0.3%, Glycerin 0.3%, Ginger root extract 0.5%, Potassium sorbate 0.2%, Sodium benzoate 0.3%, Morinda officinalis fruit extract 0.3%; Phase G: Phenoxyethanol 0.5%, Ethylhexylglycerin 0.3%, Methylparaben 0.05%, Ethylparaben 0.05%; Remaining water.

[0073] The preparation method of the Lulutong essence is as follows: Step 1: After stirring and dissolving phase A and homogenizing, keep it at 92°C for later use; Step 2: Cool down to 85℃, add phase B, and stir well; Step 3: Continue adding phase D and homogenizing, then add phase C and homogenize. Step 4: Cool down to 35℃, add phase E, phase F and phase G and stir well.

[0074] Test Example 1 Cytotoxicity test Immortalized human osteoblasts, purchased from Shanghai Jinyuan Biotechnology Co., Ltd., were seeded at a density of 5 × 10⁶ cells / year. 4 Cells were seeded per well in 96-well plates and cultured in an incubator (37°C, 5% CO2) until 80% confluence.

[0075] Remove the culture medium (DMEM medium containing 10% FBS), divide into groups, and set up 3 replicates for each group, as follows: Sample group: Add 100 μL of sample solution of different concentrations to each well (prepared with DMEM medium containing 10% FBS as solvent); Blank control group: Add 100 μL of DMEM medium containing 10% FBS to each well; Background group: Cell-free, with 100 μL of DMEM medium containing 10% FBS added to each well; The culture plate was exposed in an incubator (37℃, 5% CO2) for 48 h. The plate was then removed, the culture medium in each well was discarded, and 100 μL of 1% FBS DMEM medium was added to each well. 20 μL of MTT solution was added to each well, and the plate was incubated at 37℃ for 3 ± 0.5 h. The MTT solution was then removed, and 100 μL of DMSO was added to each well. After shaking in the dark for 10 min, the absorbance (OD) was measured at 570 nm.

[0076] Cell viability was calculated by taking the cell viability of the blank control group as 100% and calculating the relative cell viability (%) of each group.

[0077] Relative cell viability (%) = (OD sample group - OD background group) × 100% / (OD blank control group - OD background group).

[0078] The results are shown in Table 2.

[0079] Table 2

[0080] Note: 1. In the table, A represents mousse cream and B represents Shululutong essence.

[0081] The results showed that the compositions of the present invention had good safety at concentrations of 0.00055 v / v% to 0.0551 v / v%, and could be used for subsequent experiments.

[0082] Test Example 2 osteoblast differentiation alkaline phosphatase (ALP) activity Following the cell culture method of Test Example 1, cells (immortized human osteoblasts, purchased from Shanghai Jinyuan Biotechnology Co., Ltd.) were seeded into 12-well plates and cultured in an incubator for 24 hours.

[0083] Remove the culture plate, discard the culture medium in the wells, and group the samples, with 3 replicates per well, as follows: Negative control (NC): Add DMEM culture medium containing 1% FBS to each well; Model control group (MC): DMEM culture medium containing IL-1β and 1% FBS was added to each well; Sample group: Add DMEM culture medium containing IL-1β, 1% FBS and different concentrations of sample solution to each well; Positive control group (PC): Each well was filled with DMEM culture medium containing IL-1β, 1% FBS and dexamethasone; After culturing for 48 hours, the supernatant was collected and stored at -80℃. ALP activity was measured using an alkaline phosphatase assay kit.

[0084] The relative content of ALP activity in each group was calculated with the model control group as 100%.

[0085] The results are shown in Table 3.

[0086] Table 3

[0087] Note: 1. In the table, A represents mousse cream and B represents Shululutong essence; 2. # indicates a statistically significant difference compared to the MC group (P < 0.05); # indicates a statistically significant difference compared to the NC group (P < 0.05).

[0088] The results showed that the relative ALP content was significantly lower in the MC group than in the NC group (p<0.05); and the relative ALP content was significantly higher in the PC group than in the MC group (p<0.05), indicating that the model was successfully established.

[0089] Meanwhile, the relative ALP content in each A+B group was significantly higher than that in the MC group (P<0.05), indicating that the combined use of the mousse cream and Lulutong essence of the present invention has the effect of promoting ALP activity.

[0090] Test Example 3 Extracellular matrix mineralization detection Following the cell culture method of Test Example 1, cells (immortized human osteoblasts, purchased from Shanghai Jinyuan Biotechnology Co., Ltd.) were seeded into 12-well plates and cultured in an incubator for 24 hours.

[0091] Remove the culture plate, discard the culture medium in the wells, and group the samples, with 3 replicates per well, as follows: Negative control (NC): Add DMEM culture medium containing ascorbic acid and sodium β-glycerophosphate to each well; Model control group (MC): DMEM culture medium containing IL-1β, ascorbic acid and sodium β-glycerophosphate was added to each well; Sample group: Add DMEM culture medium containing IL-1β, ascorbic acid, sodium β-glycerophosphate and different concentrations of the sample to each well; Change the medium every two days and culture for 14 days. Discard the supernatant, wash once with PBS, fix with fixative for 20 minutes, and wash three times with PBS. Add 1.5 mL of Alizarin Red S staining solution to each well and stain at room temperature for 30 minutes.

[0092] Microscopic images were taken, and semi-quantitative analysis was performed using hexadecylpyridine chloride solution.

[0093] The results are shown in Table 4 and Figure 1 As shown.

[0094] Table 4

[0095] Note: 1. A is a standalone mousse cream, B is a standalone Lulutong essence, and C is a standalone zitanol; 2. # indicates a statistically significant difference compared to group MC (P < 0.05); # indicates a statistically significant difference compared to group NC (P < 0.05); ^ indicates a statistically significant difference compared to group A (P < 0.05).

[0096] The results showed that the relative expression level of extracellular matrix mineralization was significantly lower in the MC group compared with that in the NC group (P<0.05), indicating that the model was successfully established.

[0097] Meanwhile, after drug intervention, the relative expression of extracellular matrix mineralization in each group was significantly higher than that in the MC group (P<0.05); indicating that the individual mousse cream (A), Lulutong essence (B), and catalpol (C), as well as the combination of mousse cream (A) with Lulutong essence (B) and catalpol (C), all have the effect of promoting extracellular matrix mineralization.

[0098] Meanwhile, it was found that compared with group A, there were no significant differences in the relative expression levels of extracellular matrix mineralization in groups B, C, and A+C (P>0.05); however, the relative expression levels of extracellular matrix mineralization in each group A+B were significantly increased (P<0.05). This indicates that compared with mousse cream (A) alone, the combination of mousse cream (A) and Lulutong essence (B), even with a 50% reduction in dosage, still has a better effect on promoting extracellular matrix mineralization. This proves that the combination of mousse cream (A) and Lulutong essence (B) produces a synergistic effect.

[0099] Test Example 4 In vitro human chondrocyte anti-inflammatory assay Following the cell culture method of Test Example 1, cells (human chondrocytes, purchased from Shanghai Jinyuan Biotechnology Co., Ltd.) were seeded into 6-well plates and cultured in an incubator for 24 h.

[0100] Remove the culture plate, discard the culture medium in the wells, and group the samples, with 3 replicates per well, as follows: Negative control (NC): Add serum-free DMEM culture medium to each well; Model control group (MC): each well was filled with serum-free DMEM culture medium containing LPS; Sample group: Add serum-free DMEM culture medium containing LPS and different concentrations of test samples to each well; Incubate for 24 h. Remove the culture plate, collect the supernatant, store at -80℃, and measure IL-6 using an ELISA kit; add Trizol to lyse and collect the cells, store at -80℃, and detect using RT-qPCR.

[0101] The results are shown in Table 5.

[0102] Table 5

[0103] Note: 1. In the table, A is mousse cream and B is Lulutong essence; 2. # indicates a statistically significant difference compared to the MC group (P < 0.05); # indicates a statistically significant difference compared to the NC group (P < 0.05).

[0104] The results showed that compared with the NC group, the expression levels of IL-6, COX-II, and IL-1β were significantly increased in the MC group (p<0.05), indicating that the model was successfully established.

[0105] Meanwhile, after drug intervention, the relative IL-6 content in each A+B group was significantly lower than that in the MC group (P<0.05), and the expression levels of COX-II and IL-1β genes in the A+B (10:1) group and the A+B (20:1) group were significantly lower than those in the MC group (P<0.05); indicating that the combined use of mousse cream and Lulutong essence in this invention has a certain soothing effect on osteoarthritis.

[0106] Test Example 5 In vitro test of type II collagen and glycosaminoglycan content in human chondrocytes Following the cell culture method of Test Example 1, cells (human chondrocytes, purchased from Shanghai Jinyuan Biotechnology Co., Ltd.) were seeded into 6-well plates and cultured in an incubator for 24 h.

[0107] Remove the culture plate, discard the culture medium in the wells, and group the samples, with 3 replicates per well, as follows: Negative control (NC): Add serum-free DMEM culture medium to each well; Model control group (MC): each well was filled with serum-free DMEM culture medium containing LPS; Sample group: Add serum-free DMEM culture medium containing LPS and different concentrations of test samples to each well; Incubate for 24 hours. Remove the culture plate, collect the supernatant, store at -80℃, and determine the concentration using an ELISA kit. Calculate the relative concentration of COL-II for each group, with the MC group as 100%.

[0108] The results are shown in Table 6 and Figure 2As shown.

[0109] Table 6

[0110] Note: 1. A is a standalone mousse cream, B is a standalone Lulutong essence, and C is a standalone zitanol; 2. # indicates a statistically significant difference compared to group MC (P < 0.05); # indicates a statistically significant difference compared to group NC (P < 0.05); ^ indicates a statistically significant difference compared to group A (P < 0.05).

[0111] The results showed that the relative expression level of COL-II was significantly lower in the MC group compared with that in the NC group (P<0.05), indicating that the modeling was successful.

[0112] Meanwhile, after drug intervention, the relative expression level of COL-II in each group was significantly higher than that in the MC group (P<0.05); indicating that the mousse cream (A), Lulutong essence (B), and catalpol (C) alone, as well as the combination of mousse cream (A) with Lulutong essence (B) and catalpol (C), all have a certain soothing effect on osteoarthritis.

[0113] Meanwhile, it was found that compared with group A, there were no significant differences in the relative expression levels of COL-II in groups B, C, and A+C (P>0.05); however, the relative expression levels of COL-II in each of the A+B groups were significantly increased (P<0.05). This indicates that compared with mousse cream (A) alone, the combination of mousse cream (A) and Lulutong essence (B), even with a 50% reduction in dosage, still has a better soothing effect on osteoarthritis. This demonstrates that the combination of mousse cream (A) and Lulutong essence (B) produces a synergistic effect.

[0114] Test Example 6 Evaluation of isolated porcine articular cartilage-osteoarthritis model (1) Obtaining porcine articular cartilage: The femoral trochlea was fully exposed, and a cartilage block was taken from the femoral trochlea of ​​the knee joint. The cartilage block was weighed and its wet weight was recorded. The cartilage block was first soaked in HBSS at 4℃, rinsed twice, and then rinsed three times with PBS before being placed in a 24-well plate for culture.

[0115] (2) Culture of porcine articular cartilage: Incubate at 37°C and 5% CO2 for 14 days, with 2 mL of culture medium per well (DMEM containing 0.1% ITS liquid culture medium supplement). Change the medium every 3 days.

[0116] 1. Cartilage vitality screening: The cultured porcine articular cartilage was placed in a 24-well plate and cultured in an incubator for 7 days. Remove the original culture medium, add 2 mL of sample dilution solution of different concentrations to each well, and incubate in an incubator for 7 days; Remove the culture plate, collect all the supernatant from the wells, and perform the detection according to the LDH kit instructions, measuring the absorbance at a wavelength of 440 nm.

[0117] The results are shown in Table 7.

[0118] Table 7

[0119] Note: In the table, A is mousse cream and B is Lulutong essence.

[0120] The results showed that the composition had good safety at concentrations below 0.0551 v / v%, and could be used for subsequent experiments.

[0121] 2. LDH determination The cultured porcine articular cartilage was placed in a 24-well plate and cultured in an incubator for 7 days. Remove the original culture medium, divide into groups, with 8 replicates per group, as follows: Negative control (NC): Add 2 mL of DMEM culture medium containing 0.1% ITS liquid medium supplement to each well; Model control group (MC): 2 mL of DMEM culture medium containing 15 ng / mL IL-1β and 0.1% ITS liquid culture medium supplement was added to each well; Sample group: Add 2 mL of DMEM culture medium containing 15 ng / mL IL-1β, 0.1% ITS liquid culture medium supplement and different concentrations of test samples to each well.

[0122] Incubate for 7 days. Remove the culture plate, collect all supernatant from the wells, and perform the assay according to the LDH kit instructions, measuring the absorbance at 440 nm.

[0123] The test results are shown in Table 8.

[0124] Table 8

[0125] Note: 1. In the table, A is mousse cream and B is Lulutong essence; 2. # indicates a statistically significant difference compared to the MC group (P < 0.05); # indicates a statistically significant difference compared to the NC group (P < 0.05).

[0126] The results showed that the relative release of LDH was higher in the MC group than in the NC group (P<0.05), indicating that the model was successfully established.

[0127] Meanwhile, the relative release of LDH in each A+B group was significantly lower than that in the MC group (P<0.05), which can alleviate chondrocyte damage caused by inflammation.

[0128] In summary, the combination of mousse cream and Lulutong essence in this invention can effectively antagonize matrix degradation during the development of osteoarthritis, protect the integrity of the collagen fiber network, significantly reduce chondrocyte damage, effectively inhibit the degradation of cartilage matrix, and have no adverse effects on chondrocyte morphology, thus having a certain improving effect on osteoarthritis.

[0129] Test Example 7 Efficacy evaluation of animal joint dysfunction models 1. Experimental animals: SPF-grade SD rats, weighing 180±20g, male.

[0130] 2. Grouping: Based on body weight, animals were randomly divided into a blank control group (NC), a model control group (MC), a product prevention group (PR), and a product intervention group (TR), with 24 animals in each group.

[0131] 3. Modeling: Based on the concentration of the inducing agent used in the preliminary experiment, the right knee joint of SD rats under gas anesthesia was flexed backwards at 95-100° to fully open the joint space and position it at its maximum. Then, on days 1, 3, and 7, 0.2 mL of a 3% papain solution was injected; the blank control group (NC) was injected with 0.2 mL of physiological saline. From the date of modeling until before drug administration, the rats' hind paws below the ankle joint were immersed in cool water for 15 minutes daily, while simultaneously being stimulated by an electric fan; the blank control group (NC) rats were injected with 0.2 mL of physiological saline at the same site. During the modeling period, changes in rat weight, mental state, and coat color and luster were continuously observed and recorded. Administration: 0.2 g / rat was applied topically, followed by 1 minute of massage, and then 10 knee flexion and extension repetitions, 3 times a week for 4 weeks. The product prevention group (PR) began administration 2 weeks before modeling. The product is the composition prepared in Example 1, and the specific concentrations of the mousse cream (A) and the Lulutong essence (B) are (A+B=0.010v / v%+0.001v / v%).

[0132] 4. Detection indicators 4.1 Behavioral Changes The VisuGait gait analysis system was used to perform a complete gait analysis on each group of SD rats.

[0133] 4.2 ELISA detection After the experiment, rats were euthanized by spinal dislocation, and joint tissues were collected and flash-frozen in liquid nitrogen. The joint and skin tissues were then dry-milled in a cryogenic grinder, and the corresponding volume of PBS (at a weight-to-volume ratio of 1:9) was added and mixed thoroughly. The homogenate was then sonicated. Finally, the homogenate was centrifuged at 5000×g for 10 min, and the supernatant was collected for analysis. Detection indicators: Substance P, calcitonin gene-related peptide (CGRP), and IL-6.

[0134] 5. Test Results 5.1 Behavioral Changes Behavioral change detection results such as Figure 3 As shown.

[0135] 5.2 ELISA Detection (1) Relevant detection indicators in joint tissues The results are as follows Figure 4 As shown.

[0136] (2) Relevant detection indicators in skin tissue The results are as follows Figure 5 As shown.

[0137] The results show: VisuGait gait analysis showed that the product prevention group (PR) and product intervention group (TR) significantly improved the hind limb stride width and average footprint area in SD rats with rheumatic knee osteoarthritis.

[0138] ELISA results showed that the product prevention group (PR) and product intervention group (TR) significantly reduced the levels of local joint neuropeptide (CGRP), substance P (SP), and inflammatory cytokine IL-6 in the joint tissue of SD rats with rheumatoid arthritis of the knee (P < 0.05). Simultaneously, the product prevention group (PR) and product intervention group (TR) also significantly reduced the level of inflammatory cytokine IL-6 in the skin tissue at the treatment site (P < 0.05).

[0139] In summary, this invention provides a topical composition and pharmaceutical preparation with clearly defined efficacy targets, outstanding synergistic advantages, and good safety. It is an important supplement and technological optimization to existing joint sub-health protection products and has significant clinical application value.

[0140] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A composition for protecting against joint sub-health, characterized in that, It contains a mousse cream and a lulutong essence in a mass ratio of 10-40:1; The mousse cream, by weight percentage, comprises: Phase A: Water, propylene glycol 1%-5%, butylene glycol 1%-3%, disodium EDTA 0.01%-0.1%, acetylglucosamine 0.1%-1.0%, sodium chondroitin sulfate 0.01%-0.5%, lycopodium extract 0.01%-0.5%, and Luffa cylindrica extract 0.01%-0.5%; Phase B: Dipropylene glycol 1.0%-10.0%, sodium polyacrylate grafted starch 0.1%-1.0%; Phase C: Rice bran oil 1.0%-10.0%, Caprylic / Capric triglycerides 1.0%-10.0%, Isopropyl palmitate 1.0%-5.0%, Hydrogenated polyisobutylene 1.0%-4.0%, Isocerocetyl ether-20 0.1%-1%, Methyl glucetyl ether-20 0.1%-1%, Glyceryl stearate 0.1%-1%, Polydimethylsiloxane 0.5%-2.0%, Cyclopentasiloxane 0.1%-0.5%, Cyclohexylsiloxane 0.1%-0.5%, Sorbitan oleate 0.1%-0.5%, Cetearyl alcohol 0.1%-0.5%, Potassium azeloyl diglycinate 0.1%-0.5%; Phase D: Tocopherol 0.05%-0.1%, Sunflower seed oil 0.05%-0.1%; Phase E: Tetrahydropiperine 0.01%-0.1%, Propylene glycol 0.05%-1.0%; Phase F: Phenoxyethanol 0.1%-1%, Ethylhexylglycerin 0.1%-1%, Methylparaben 0.05%-0.1%, Ethylparaben 0.05%-0.1%; Phase G: Gentiana macrophylla 0.1%-2%; H phase: 0.01%-0.1% white chrysanthemum flower oil, 0.01%-0.1% rose flower oil, 0.01%-0.1% cinnamon leaf oil, 0.01%-0.14% ylang-ylang flower oil, 0.001%-0.1% ginger root oil, 0.01%-0.2% vanillyl butyl ether, 0.01%-0.2% kava pepper, 0.01%-0.3% white beech leaf oil; Remaining water; The Lulutong essence, by mass percentage, comprises: Phase A: Water, propylene glycol 1%-15%, magnesium aluminum silicate 0.05%-0.3%, sodium chondroitin sulfate 0.01%-0.5%, lycopodium extract 0.01%-0.2%, and Luffa cylindrica extract 0.01%-0.2%; Phase B: Sodium stearoyl glutamate 0.10%-0.2%, lauryl ether-3 0.5%-3%, sorbitan sesquioleate 0.1%-1%, sorbitan trioleate 0.1%-1%, sorbitan stearate 0.1%-0.5%; Phase C: 0.1%-1% acrylic (ester) copolymers, 0.1%-0.5% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 0.10%-0.5% isohexadecane, 0.01%-0.5% polysorbate-60, 0.01%-0.5% sorbitan isostearate; Phase D: Frankincense oil 0.1%-1%, methyl salicylate 0.1%-1%, Ceylon cinnamon leaf oil 0.1%-1%, lemongrass leaf oil 0.1%-1%, vanillyl butyl ether 0.05%-1.5%, camphor 0.1%-4%, menthol 0.1%-1.5%, butylated hydroxytoluene 0.01%-0.2%, tetrahydropiperine 0.01%-0.2%; Phase E: Triethanolamine 0.05%-0.2%, Citric Acid 0.01%-0.2%; Phase F: 1,2-Pentanediol 0.10%-0.5%, Butylene glycol 0.1%-2%, Hydroxyphenylpropionamide benzoic acid 0.01%-0.5%, Ascorbyl palmitate 0.1%-0.5%, Glycerin 0.1%-5%, Ginger root extract 0.01%-1%, Potassium sorbate 0.01%-0.5%, Sodium benzoate 0.01%-0.5%, Morinda officinalis fruit extract 0.01%-0.5%; Phase G: Phenoxyethanol 0.1%-1%, Ethylhexylglycerin 0.1%-0.5%, Methylparaben 0.01%-0.1%, Ethylparaben 0.01%-0.1%; Remaining water.

2. The composition according to claim 1, characterized in that, The mass ratio of the mousse cream to the Lulutong essence is 10-20:

1.

3. The composition according to claim 1, characterized in that, The mass ratio of the mousse cream to the Lulutong essence is 10:

1.

4. The composition according to claim 1, characterized in that, The mousse cream, by weight percentage, comprises: Phase A: Water, propylene glycol 2.0-4.0%, butylene glycol 1.5-2.5%, disodium EDTA 0.03-0.07%, acetylglucosamine 0.2%-0.8%, sodium chondroitin sulfate 0.15%-0.35%, lycopodium extract 0.15%-0.35%, and Luffa cylindrica extract 0.1%-0.3%; Phase B: Dipropylene glycol 3.0%-7.0%, sodium polyacrylate grafted starch 0.15%-0.5%; Phase C: Rice bran oil 3.0%-9.0%, caprylic / capric triglycerides 5%-10%, isopropyl palmitate 2.0%-4.0%, hydrogenated polyisobutylene 1.0%-3.0%, isocetyl alcohol polyether-20 0.3%-0.7%, methyl glucetol polyether-20 0.1%-0.5%, glyceryl stearate 0.4%-0.8%, polydimethylsiloxane 0.5%-1.5%, cyclopentamethoxydimethylsiloxane 0.2%-0.4%, cyclohexylsiloxane 0.2%-0.4%, sorbitan oleate 0.2%-0.4%, cetearyl alcohol 0.2%-0.4%, potassium azeloyl diglycinate 0.2%-0.4%; Phase D: Tocopherol 0.06%-0.1%, Sunflower seed oil 0.06%-0.1%; Phase E: Tetrahydropiperine 0.06%-0.1%, Propylene glycol 0.06%-0.1%; Phase F: Phenoxyethanol 0.3%-0.5%, Ethylhexylglycerin 0.03%-0.5%, Methylparaben 0.05%-0.08%, Ethylparaben 0.05%-0.08%; Phase G: Gentiana macrophylla 0.5%-1.5%; H phase: 0.03%-0.07% white chrysanthemum flower oil, 0.03%-0.07% rose flower oil, 0.03%-0.07% cinnamon leaf oil, 0.03%-0.10% ylang-ylang flower oil, 0.06%-0.09% ginger root oil, 0.05%-0.12% vanillyl butyl ether, 0.4%-1.4% kava pepper, 0.15%-0.25% white beech leaf oil; Remaining water.

5. The composition according to claim 1, characterized in that, The mousse cream, by weight percentage, comprises: Phase A: Water, propylene glycol 3.0%, butylene glycol 2.0%, disodium EDTA 0.05%, acetylglucosamine 0.5%, sodium chondroitin sulfate 0.25%, lycopodium extract 0.25%, and Luffa cylindrica extract 0.2%; Phase B: Dipropylene glycol 5.0%, sodium polyacrylate grafted starch 0.2%; Phase C: Rice bran oil 6.0%, Caprylic / Capric triglyceride 8.0%, Isopropyl palmitate 3.0%, Hydrogenated polyisobutylene 2.0%, Isocerocetyl ether-20 0.5%, Methyl glucetyl ether-20 0.3%, Glyceryl stearate 0.6%, Polydimethylsiloxane 1.0%, Cyclopentasiloxane 0.3%, Cyclohexylsiloxane 0.3%, Sorbitan oleate 0.3%, Cetearyl alcohol 0.3%, Potassium azeloyl diglycinate 0.3%; Phase D: Tocopherol 0.08%, Sunflower Seed Oil 0.08%; Phase E: Tetrahydropiperine 0.08%, Propylene glycol 0.08%; Phase F: Phenoxyethanol 0.5%, Ethylhexylglycerin 0.5%, Methylparaben 0.08%, Ethylparaben 0.08%; G phase: Gentiana macrophylla 1.0%; H phase: 0.05% white chamomile flower oil, 0.05% rose flower oil, 0.05% cinnamon leaf oil, 0.07% ylang-ylang flower oil, 0.08% ginger root oil, 0.08% vanillyl butyl ether, 0.9% kava pepper, 0.2% flat-leaved white beech leaf oil; Remaining water.

6. The composition according to claim 1, characterized in that, The method for preparing the mousse cream includes the following steps: Step 1: Stir and dissolve phase A, homogenize and keep warm, then set aside; Step 2: Cool down, add phase C and phase D, and stir well; Step 3: Continue adding phase B and homogenizing, then add phase E and homogenize. Step 4: Cool down to below 45℃, add phase F, phase G and phase H and stir well.

7. The composition according to claim 1, characterized in that, The Lulutong essence, by mass percentage, comprises: Phase A: Water, propylene glycol 8.0%-12.0%, magnesium aluminum silicate 0.05%-0.15%, sodium chondroitin sulfate 0.2%-0.4%, lycopodium extract 0.05%-0.15%, and Luffa cylindrica extract 0.05%-0.15%; Phase B: Sodium stearoyl glutamate 0.12%-0.18%, lauryl ether-3 1.5%-2.5%, sorbitan sesquioleate 0.3%-0.7%, sorbitan trioleate 0.3%-0.7%, sorbitan stearate 0.2%-0.4%; Phase C: 0.5%-0.9% acrylic (ester) copolymers, 0.2%-0.4% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 0.2%-0.4% isohexadecane, 0.25%-0.35% polysorbate-60, and 0.2%-0.4% sorbitan isostearate. Phase D: Frankincense oil 0.2%-0.8%, methyl salicylate 0.2%-0.8%, Ceylon cinnamon leaf oil 0.2%-0.8%, lemongrass leaf oil 0.2%-0.8%, vanillyl butyl ether 0.07%-1.2%, camphor 1%-3%, menthol 0.8%-1.2%, butylated hydroxytoluene 0.05%-0.15%, tetrahydropiperine 0.05%-0.15%; Phase E: Triethanolamine 0.05%-0.15%, Citric Acid 0.05%-0.15%; Phase F: 1,2-Pentanediol 0.10%-0.3%, Butylene glycol 0.5%-2%, Hydroxyphenylpropionamide benzoic acid 0.1%-0.3%, Ascorbyl palmitate 0.2%-0.4%, Glycerin 0.2%-0.5%, Ginger root extract 0.2%-0.8%, Potassium sorbate 0.1%-0.3%, Sodium benzoate 0.1%-0.3%, Morinda officinalis fruit extract 0.2%-0.4%; Phase G: Phenoxyethanol 0.2%-0.5%, Ethylhexylglycerin 0.1%-0.3%, Methylparaben 0.02%-0.05%, Ethylparaben 0.02%-0.05%; Remaining water.

8. The composition according to claim 1, characterized in that, The Lulutong essence, by mass percentage, comprises: Phase A: Water, 10% propylene glycol, 0.1% magnesium aluminum silicate, 0.3% sodium chondroitin sulfate, 0.1% lycopodium extract, 0.1% luffa extract; Phase B: Sodium stearoyl glutamate 0.15%, lauryl ether-3 2%, sorbitan sesquioleate 0.5%, sorbitan trioleate 0.5%, sorbitan stearate 0.3%; Phase C: 0.7% acrylic (ester) copolymer, 0.3% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 0.3% isohexadecane, 0.3% polysorbate-60, 0.3% sorbitan isostearate; Phase D: Frankincense oil 0.5%, methyl salicylate 0.5%, Ceylon cinnamon leaf oil 0.5%, lemongrass leaf oil 0.5%, vanillyl butyl ether 1%, camphor 2%, menthol 1%, butylated hydroxytoluene 0.1%, piperine 0.1%; Phase E: Triethanolamine 0.1%, Citric Acid 0.1%; Phase F: 1,2-Pentanediol 0.2%, Butylene glycol 1%, Hydroxyphenylpropionamide benzoic acid 0.2%, Ascorbyl palmitate 0.3%, Glycerin 0.3%, Ginger root extract 0.5%, Potassium sorbate 0.2%, Sodium benzoate 0.3%, Morinda officinalis fruit extract 0.3%; Phase G: Phenoxyethanol 0.5%, Ethylhexylglycerin 0.3%, Methylparaben 0.05%, Ethylparaben 0.05%; Remaining water.

9. The composition according to claim 1, characterized in that, The preparation method of the Lulutong essence includes the following steps: Step 1: After stirring and dissolving phase A and homogenizing it, keep it warm for later use; Step 2: Cool down, add phase B, and stir well; Step 3: Continue adding phase D and homogenizing, then add phase C and homogenize. Step 4: Cool down to below 45℃, add phase E, phase F and phase G and stir well.

10. A topical preparation for protecting against joint sub-health, characterized in that, Includes the composition according to any one of claims 1-8.

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