Application of small chemical molecules in prevention or treatment of obesity

Abstract

The invention relates to application of small chemical molecules in prevention or treatment of obesity and a composition containing the small chemical molecules, in particular to vitamin D, 25-hydroxyvitamin D or a vitamin D receptor agonist and application of analogues of the substances in preparation of products. Wherein the product is used for preventing and / or alleviating or treating obesity, overweight, or a symptom or state or trend or disease or condition caused by or associated with overweight or obesity in a mammal, or for reducing body weight.

Description

TECHNICAL FIELD

[0001] The present application belongs to the new use of vitamin D, 25-hydroxyvitamin D, vitamin D receptor agonists and analogs of the above-mentioned substances, compositions containing them and their methods for preventing and / or treating obesity and obesity-related symptoms or diseases. BACKGROUND

[0002] At present, obesity has become a global problem of public health. Obesity can be defined as an abnormal state of excessive accumulation or abnormal distribution of fat in the body, which leads to the impact on physical health. Obesity is mainly classified according to the body mass index (BMI). In addition, obesity is also accompanied by other diseases, such as hypertension, cardiovascular disease, diabetes, respiratory disease, non-alcoholic fatty liver disease, breast cancer, colorectal cancer, hepatocellular carcinoma, infertility, etc.

[0003] The World Health Organization considers BMI of 25.0 kg / m 2 or higher, body mass index of 30.0 kg / m2and above, as obesity. When the body mass index is about 21.0 kg / m 2 2, the risk of diabetes, hypertension and dyslipidemia will increase, thereby reducing the life expectancy and greatly increasing the individual health and social economic burden. At present, overweight has become the sixth largest risk factor for global disease burden.

[0004] At present, the main methods to achieve weight loss are lifestyle intervention, drug or small molecule weight loss, and obesity surgery. Obesity surgery has an immediate effect and long-term effect, but the surgery itself has a high risk and may have complications after surgery. In addition to obesity surgery, other weight loss methods currently available essentially involve reducing energy intake and increasing energy consumption, which are achieved by changing cell size and are difficult to have long-term effect. Therefore, how to achieve long-term weight loss effect by a low-risk method is a scientific problem to be solved in the field.

[0005] Vitamin D is a fat-soluble prohormone, a general term for a group of structurally similar sterol derivatives with anti-rickets effect, a safe natural product, and a necessary nutrient for the human body. One of the main functions of vitamin D is to maintain the levels of calcium and phosphorus in the blood within a healthy physiological range, thereby maintaining various metabolic functions, transcriptional regulation, and bone metabolism. Cholecalciferol (vitamin D3) and ergocalciferol (vitamin D2) are the ones that really play a role in animal nutrition and play an important role in various tissues and organs in the body. Vitamin D3 is the main source of vitamin D for animal individuals, most of which is endogenously produced in the skin after sun exposure, and a small part is derived from food; while vitamin D2 mainly exists in plants and is a non-animal form of vitamin D. Once inside the body, vitamin D3 and vitamin D2 are converted to 25-hydroxyvitamin D in the liver tissue under the action of vitamin D-25-hydroxylase, and 25-hydroxyvitamin D can be further converted to 1, 25-dihydroxyvitamin D with in vivo activity in the kidney by 25-hydroxyvitamin D-1α-hydroxylase. 1, 25-dihydroxyvitamin D is a vitamin D receptor (VDR) agonist that can bind to it to function, including inhibiting cell proliferation and inducing terminal differentiation, inhibiting angiogenesis, stimulating insulin production, inducing apoptosis, inhibiting renin production, and stimulating endogenous antimicrobial peptide production in macrophages.

[0006] Therefore, there is still an urgent need in human society to develop simpler and safer substances to solve obesity and the health problems caused by obesity. SUMMARY

[0007] The present inventors unexpectedly found in the research that intervention of obese mice with vitamin D, 25-hydroxyvitamin D, vitamin D receptor agonists, and analogs of the above substances can significantly reduce the body weight of obese mice and the body weight will not rebound after stopping intervention. Specifically, high-fat diet-induced wild-type C57BL / 6N obese mice were used as research objects in the present study, which simulated most obese people under natural conditions. Vitamin D3 (VD3) was used to treat high-fat diet-induced obese mice, and the body weight of high-fat diet-induced obese mice was reduced by an average of 34.6% and a maximum of 50% within four weeks; the body fat content was reduced by an average of 61.3% and a maximum of 81.7%. It can be proved that vitamin D3 short-term treatment can have a very significant weight loss effect, which is characterized by fast effect and large weight and body fat reduction.

[0008] It is well known that obesity leads to the decline of many functions. This study not only proves that vitamin D3 has a significant weight loss effect, but also proves that vitamin D3 can restore the decline of motor balance function caused by obesity in high-fat diet-induced obese mice through the rotarod test; through the endurance test, it is proved that vitamin D3 can significantly improve the endurance of obese mice; through the glucose tolerance test, it is proved that vitamin D3 can significantly improve the blood glucose regulation function of obese mice. Furthermore, this study proves that using vitamin D3 to treat mice at the same time as high-fat diet, the mice can maintain normal weight at all times, and has the effect of preventing obesity. More excitingly, this study found that after four weeks of vitamin D3 treatment while the mice were on a high-fat diet, the mice continued to have a weight loss effect for at least three months after the vitamin D3 treatment stopped. Therefore, vitamin D3 has a long-term weight loss effect.

[0009] In addition, this study comprehensively evaluates the long-term effect of small molecules in vivo: on the one hand, the survival rate of the experimental group mice is higher than that of the control group mice, and the life span is longer after more than one year of long-term observation of the mice treated with small molecules. On the other hand, through histological detection of multiple tissue organs, no obvious abnormalities were found in the multiple tissue organs of the experimental group mice; at the same time, compared with the fatty liver condition of the control group, the experimental group mice had obvious improvement. The experimental results of these two aspects exclude the toxic effect of small molecules.

[0010] This study also explores the weight loss mechanism of vitamin D3 and excludes the possibility that vitamin D3 mainly achieves weight loss effect by greatly limiting energy intake. At the same time, this study clearly shows that high-dose vitamin D3 is specifically enriched in adipose tissue in vivo, resulting in a specific reduction in the number of adipocytes, thereby achieving weight loss. Therefore, from the perspective of the mechanism of small molecule action, the safety of its weight loss effect is further demonstrated. This is completely different from other small molecules and other weight loss methods that have been reported, and is a completely new mechanism of action. Through transcriptome analysis of the liver tissue of the mice, this study found that the fatty acid degradation and lipid metabolism-related signaling pathways in the liver tissue treated with vitamin D3 were activated, which to some extent explained the effect of improving fatty liver symptoms.

[0011] The present invention relates to the use of vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonists and analogs of the above-mentioned substances for the preparation of a product for the prevention and / or alleviation or treatment of obesity, overweight, or a symptom or state or tendency or disease or condition caused by or associated with overweight or obesity in a mammal, or for weight loss.

[0012] The present invention relates to a product for preventing and / or alleviating or treating obesity, overweight, or a symptom or condition or tendency or disease or disorder caused by or associated with overweight or obesity, or for weight loss in a mammal, comprising a weight loss or anti-obesity effective amount of vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist, and analogs of the foregoing, any excipient or carrier permitted for use in commerce.

[0013] The present invention relates to a product for preventing and / or alleviating or treating obesity, overweight, or a symptom or condition or tendency or disease or disorder caused by or associated with overweight or obesity, or for weight loss in a mammal, comprising a weight loss or anti-obesity effective amount of vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist, and analogs of the foregoing.

[0014] The present invention also relates to a method for preventing and / or alleviating or treating obesity, overweight, or a symptom or condition or tendency or disease or disorder caused by or associated with overweight or obesity in a mammal, comprising administering to a subject in need of preventing and / or alleviating or treating obesity, overweight, or a symptom or condition or tendency or disease or disorder caused by or associated with overweight or obesity in a mammal, an effective amount of vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist, and analogs of the foregoing.

[0015] According to the present invention, an "effective amount" means any amount of vitamin D, 25-hydroxyvitamin D, or a vitamin D receptor agonist, and analogs of the foregoing, which when used in a mammal, produces the desired effect of preventing and / or alleviating or treating obesity, overweight, or a symptom or condition or tendency or disease or disorder caused by or associated with overweight or obesity in a mammal. For example, when the vitamin D, 25-hydroxyvitamin D, or a vitamin D receptor agonist, and analogs of the foregoing, is administered parenterally to a mammal, e.g., intravenously, intraperitoneally, subcutaneously, an effective amount is at least 1 mg per week (for humans) or at least 0.1 mg (for example, for common pets such as cats, dogs, etc.) or at least 0.015 mg (for example, for small mammals such as mice, etc.). When the vitamin D, 25-hydroxyvitamin D, or a vitamin D receptor agonist, and analogs of the foregoing, is administered enterally to a mammal, e.g., orally, an effective amount is at least 1 mg per week (for humans) or at least 0.1 mg (for example, for common pets such as cats, dogs, etc.) or at least 0.015 mg (for example, for small mammals such as mice, etc.).

[0016] In the present application, when the subject is a human, the administration dose of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above is at least 1 mg per week, for example, 1-40 mg, 1-35 mg, 1-30 mg, 1-28 mg, 1-25 mg, 1-22 mg, 1-20 mg, 1-18 mg, 1-15 mg, 1-12 mg, 1-10 mg, 1-9 mg, 1-8 mg, 1-7 mg, 1-6 mg, 1-5 mg, 2-50 mg, 2-40 mg, 2-30 mg, 2-25 mg, 2-22 mg, 2-20 mg, 2-18 mg, 1-15 mg, 2-12 mg, 2-10 mg, 2-8 mg, 2-6 mg, 2-5 mg, 3-50 mg, 3-40 mg, 3-30 mg, 3-25 mg, 3-20 mg, 3-18 mg, 3-15 mg, 3-12 mg, 3-10 mg, 3-8 mg, 3-7 mg, 3-6 mg, 3-5 mg, 4-50 mg, 4-40 mg, 4-35 mg, 4-30 mg, 4-25 mg, 4-20 mg, 4-18 mg, 4-15 mg, 4-12 mg, 4-10 mg, 4-8 mg, 4-6 mg, 4-5 mg, 5-50 mg, 5-40 mg, 5-35 mg, 5-30 mg, 5-25 mg, 5-22 mg, 5-20 mg, 5-18 mg, 5-15 mg, 5-12 mg, 5-10 mg, 5-8 mg, 5-6 mg. The administration period of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above is more than 7 days, for example, 7-60 days.

[0017] mg, 1-80 mg, 1-60 mg, 1-40 mg, 1-30 mg, 1-20 mg, 1-10 mg, 2-350 mg, 2-300 mg, 2-200 mg, 2-100 mg, 2-50 mg, 2-30 mg, 3-350 mg, 3-300 mg, 3-200 mg, 3-150 mg, 3-100 mg, 3-80 mg, 3-60 mg, 3-40 mg, 3-30 mg, 4-350 mg, 4-320 mg, 4-300 mg, 4-280 mg, 4-250 mg, 4-230 mg, 4-200 mg, 4-180 mg, 4-150 mg, 4-130 mg, 4-100 mg, 4-80 mg, 4-60 mg, 4-40 mg, 4-30 mg, 6-350 mg, 6-320 mg, 6-300 mg, 6-280 mg, 6-250 mg, 6-220 mg, 6-200 mg, 6-180 mg, 6-150 mg, 6-120 mg, 6-100 mg, 6-80 mg, 6-60 mg, 6-50 mg, 6-40 mg, 6-30 mg, 6-20 mg, 8-350 mg, 8-320 mg, 8-300 mg, 8-280 mg, 8-250 mg, 8-220 mg, 8-200 mg, 8-180 mg, 8-150 mg, 8-120 mg, 8-100 mg, 8-90 mg, 8-80 mg, 8-60 mg, 8-50 mg, 8-40 mg, 8-30 mg, 8-20 mg, 10-300 mg, 10-250 mg, 10-200 mg, 10-150 mg, 10-100 mg, 10-80 mg, 10-60 mg, 10-40 mg, 10-30 mg, 15-300 mg, 15-250 mg, 15-200 mg, 15-150 mg, 15-100 mg, 15-80 mg, 15-60 mg, 15-40 mg, 15-25 mg, 20-300 mg, 20-250 mg, 20-200 mg, 20-150 mg, 20-100 mg, 20-80 mg, 20-60 mg, 20-40 mg, 30-300 mg, 30-250 mg, 30-200 mg, 30-100 mg, 30-80 mg, 40-300 mg, 40-250 mg, 40-200 mg, 40-150 mg, 40-100 mg, 40-80 mg, 40-60 mg.

[0018] In the present application, when the subject is a non-human mammal such as a cat, a dog, the administration dose of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and the analogs of the above substances is 0.1 mg or more per week, for example, 0.1 to 10 mg, 0.1 to 9 mg, 0.1 to 8 mg, 0.1 to 7 mg, 0.1 to 6 mg, 0.1 to 5 mg, 0.1 to 4 mg, 0.1 to 3 mg, 0.1 to 2 mg, 0.1 to 1 mg, 0.1 to 0.8 mg, 0.1 to 0.6 mg, 0.1 to 0.4 mg, 0.1 to 0.2 mg, 0.2 to 10 mg, 0.2 to 9 mg, 0.2 to 8 mg, 0.2 to 7 mg, 0.2 to 6 mg, 0.2 to 5 mg, 0.2 to 4 mg, 0.2 to 3 mg, 0.2 to 2 mg, 0.2 to 1 mg, 0.2 to 0.8 mg, 0.2 to 0.6 mg, 0.2 to 0.4 mg, 0.4 to 10 mg, 0.4 to 9 mg, 0.4 to 8 mg, 0.4 to 6 mg, 0.4 to 5 mg, 0.4 to 4 mg, 0.4 to 3 mg, 0.4 to 2 mg, 0.4 to 1 mg, 0.4 to 0.8 mg, 0.4 to 0.6 mg, 0.5 to 10 mg, 0.5 to 8 mg, 0.5 to 6 mg, 0.5 to 5 mg, 0.5 to 4 mg, 0.5 to 3 mg, 0.5 to 2 mg, 0.5 to 1 mg, 0.6 to 10 mg, 0.6 to 8 mg, 0.6 to 6 mg, 0.6 to 4 mg, 0.8 to 10 mg, 0.8 to 8 mg, 0.8 to 6 mg, 0.8 to 5 mg, 0.8 to 4 mg, 0.8 to 3 mg, 0.8 to 2 mg, 1 to 10 mg, 1 to 9 mg, 1 to 8 mg, 1 to 6 mg, 1 to 5 mg, 1 to 4 mg, 1 to 3 mg, 1 to 2 mg, 2 to 10 mg, 2 to 8 mg, 2 to 6 mg, 2 to 5 mg, 2 to 4 mg, 2 to 3 mg, 3 to 10 mg, 3 to 8 mg, 3 to 6 mg, 3 to 5 mg. The administration period of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and the analogs of the above substances is 7 days or more, for example, 7 to 60 days.

[0019] Thus, when the subject is a non-human mammal, such as a cat, dog, the effective amount of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above are at least 0.1 mg, such as 0.1-80 mg, 0.1-70 mg, 0.1-60 mg, 0.1-50 mg, 0.1-40 mg, 0.1-30 mg, 0.1-20 mg, 0.1-10 mg, 0.1-8 mg, 0.1-6 mg, 0.1-4 mg, 0.1-2 mg, 0.3-80 mg, 0.3-60 mg, 0.3-40 mg, 0.3-20 mg, 0.3-10 mg, 0.3-5 mg, 0.3-3 mg, 0.5-80 mg, 0.5-60 mg, 0.5-40 mg, 0.5-20 mg, 0.5-10 mg, 0.5-8 mg, 0.5-6 mg, 0.5-4 mg, 0.8-80 mg, 0.8-60 mg, 0.8-40 mg, 0.8-20 mg, 0.8-10 mg, 0.8-6 mg, 0.8-4 mg, 1-80 mg, 1-60 mg, 1-40 mg, 1-20 mg, 3-80 mg, 3-60 mg, 3-40 mg, 3-20 mg, 3-10 mg, 4-80 mg, 4-60 mg, 4-40 mg, 4-20 mg, 5-80 mg, 5-60 mg, 5-40 mg, 5-20 mg, 5-10 mg.

[0020] In the present application, when the subject is a non-human mammal such as a mouse, a rat, and the like, a small mammal, the administration dose of the vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist, and the like of the above-mentioned substance is 0.015 mg or more per week, for example, 0.015 to 1 mg, 0.015 to 0.8 mg, 0.015 to 0.6 mg, 0.015 to 0.4 mg, 0.015 to 0.2 mg, 0.015 to 0.1 mg, 0.015 to 0.08 mg, 0.015 to 0.06 mg, 0.015 to 0.03 mg, 0.015 to 0.02 mg, 0.03 to 1 mg, 0.03 to 0.8 mg, 0.03 to 0.6 mg, 0.03 to 0.4 mg, 0.03 to 0.2 mg, 0.03 to 0.1 mg, 0.03 to 0.08 mg, 0.03 to 0.06 mg, 0.03 to 0.04 mg, 0.05 to 1 mg, 0.05 to 0.8 mg, 0.05 to 0.6 mg, 0.05 to 0.5 mg, 0.05 to 0.4 mg, 0.05 to 0.3 mg, 0.05 to 0.1 mg, 0.05 to 0.08 mg, 0.05 to 0.06 mg, 0.1 to 1 mg, 0.1 to 0.8 mg, 0.1 to 0.6 mg, 0.1 to 0.4 mg, 0.1 to 0.3 mg, 0.3 to 1 mg, 0.3 to 0.6 mg, 0.3 to 0.5 mg, 0.4 to 1 mg, 0.4 to 0.8 mg, 0.4 to 0.6 mg, 0.6 to 1 mg, 0.6 to 0.8 mg. The administration period of the vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist, and the like of the above-mentioned substance is 7 days or more, for example, 7 to 60 days.

[0021] Thus, when the subject is a non-human mammal, such as a mouse, rat, or other small mammal, the effective amount of the vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist, and analogs of the foregoing, is at least 0.015 mg, such as 0.015-8 mg, 0.015-6 mg, 0.015-4 mg, 0.015-3 mg, 0.015-2 mg, 0.015-1 mg, 0.015-0.8 mg, 0.015-0.5 mg, 0.015-0.1 mg, 0.03-8 mg, 0.03-6 mg, 0.03-4 mg, 0.03-2 mg, 0.03-1 mg, 0.03-0.07 mg, 0.03-0.05 mg, 0.05-8 mg, 0.05-6 mg, 0.05-4 mg, 0.05-2 mg, 0.05-1 mg, 0.05-0.08 mg, 0.08-8 mg, 0.08-6 mg, 0.08-4 mg, 0.08-2 mg, 0.1-8 mg, 0.1-6 mg, 0.1-4 mg, 0.1-2 mg, 1-8 mg, 1-6 mg, 1-4 mg, 1-3 mg, 2-8 mg, 2-6 mg, 2-5 mg, 2-4 mg, 3-8 mg, 3-6 mg, 3-5 mg, 4-8 mg, 4-6 mg, 5-8 mg.

[0022] In the present application, the effective amount can be in a single dosage form, or in multiple dosage forms (as long as the sum of the multiple dosage forms is the effective amount); for example, when the effective amount is 4 mg, it can be in one 4 mg dosage form; in two 2 mg dosage forms; or in one 2 mg and two 1 mg dosage forms.

[0023] According to the present application, the "product" can be any form suitable for use of the vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist, and analogs of the foregoing, such as a human food, a human drug, an animal food, or an animal drug, or a kit.

[0024] According to the present application, the vitamin D and analogs thereof include, but are not limited to, vitamin D2, vitamin D3, vitamin D4, (R)-vitamin D2, 11-hydroxymeridinol, 1α-hydroxy vitamin D4; preferably vitamin D3

[0025] According to the present application, the 25-hydroxyvitamin D and analogs thereof include, but are not limited to, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 25,26-dihydroxyvitamin D3, 5,6-trans vitamin D2, 25-hydroxyvitamin D3 monohydrate.

[0026] According to the present application, the vitamin D receptor agonist includes, but is not limited to, 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D2, trans calcitriol.

[0027] According to the present application, the product can be in the form of product packaging, product labeling, product content, and product instructions. The product instructions can indicate that the vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist and analogs of the above substances are administered at a dose of 0.1-100 mg / kg, or the effective amount of the vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist and analogs of the above substances is at least 1 mg (for humans) or at least 0.1 mg (for example, for cats, dogs) or at least 0.015 mg (for example, for mice, rats), and optionally the product instructions indicate that the administration frequency is 1, 2, 3, or 7 times per week, for example, once every 2 or 3 days, and the administration cycle is 7 days or more, for example, 7-60 days.

[0028] According to the present application, the mammal includes, but is not limited to, humans, dogs, cats, mice, rats, and the like.

[0029] According to the present application, the symptoms or states or trends or diseases or conditions caused or associated with overweight or obesity include, but are not limited to, physical discomfort caused by overweight or obesity, elevated physical indicators of certain future possible diseases caused by overweight or obesity, future morbidity risk possibly brought by family history, diabetes, fatty liver, high blood pressure.

[0030] According to the present application, any excipient or carrier allowed for use in the market can be any excipient or carrier allowed for use in the market, for example, at least one of a sustained-release agent, an excipient, a filler, a binder, a humectant, a disintegrant, an absorption enhancer, a surfactant, and a lubricant.

[0031] The present application also relates to the use of the vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist and analogs of the above substances for inducing apoptosis of adipocytes.

[0032] The present application also relates to a method for inducing apoptosis of adipocytes in a sample or a subject, which comprises adding vitamin D, 25-hydroxyvitamin D, or a vitamin D receptor agonist and analogs of the above substances to the sample or the subject.

[0033] Specifically, the present application provides the following technical solutions:

[0034] 1. Use of vitamin D, 25-hydroxyvitamin D or a vitamin D receptor agonist and analogs of the above in the manufacture of a product for preventing and / or alleviating or treating obesity, overweight, or a symptom or state or tendency or disease or condition caused by or associated with overweight or obesity, or for weight loss in a mammal.

[0035] 2. Use according to item 1, wherein the effective amount of the vitamin D, 25-hydroxyvitamin D or a vitamin D receptor agonist and analogs of the above is at least 1 mg (for humans) or at least 0.1 mg (e.g. for cats, dogs) or at least 0.015 mg (e.g. for mice, rats).

[0036] 3. Use according to item 1 or 2, wherein the vitamin D and analogs thereof include, but are not limited to, vitamin D2, vitamin D3, vitamin D4, (R)-vitamin D2, 11-hydroxymeridinol, 1a-hydroxyvitamin D4; preferably vitamin D3;

[0037] the 25-hydroxyvitamin D and analogs thereof include, but are not limited to, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 25,26-dihydroxyvitamin D3, 5,6-trans vitamin D2, 25-hydroxyvitamin D3 monohydrate;

[0038] the vitamin D receptor agonist includes, but is not limited to, 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D2, trans calcitriol.

[0039] 4. Use according to any one of items 1-3, wherein the vitamin D, 25-hydroxyvitamin D or a vitamin D receptor agonist and analogs of the above are in a form suitable for intravenous, intraperitoneal, subcutaneous or oral administration.

[0040] 5. Use according to any one of items 1-4, wherein the vitamin D, 25-hydroxyvitamin D or a vitamin D receptor agonist and analogs of the above are administered in a dose of 0.1-100 mg / kg in a non-human mammal.

[0041] 6. The use of any one of items 1-5, wherein the product further comprises a product insert indicating that the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the foregoing are to be administered at a dose of 0.1-100 mg / kg, or indicating that an effective amount of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the foregoing is at least 1 mg (for humans) or at least 0.1 mg (e.g., for cats, dogs) or at least 0.015 mg (e.g., for mice, rats), optionally the product insert indicating that the frequency of administration is 1, 2, 3, or 7 times per week, with a cycle of administration of 7 days or more, e.g., 7-60 days.

[0042] 7. The use of any one of items 1-6, wherein the mammal comprises: a human, a dog, a cat, a mouse, a rat.

[0043] 8. The use of any one of items 1-7, wherein the condition or state or tendency or disease or disorder caused by or associated with excess weight or obesity comprises, but is not limited to: diabetes, fatty liver, hypertension.

[0044] 9. The use of any one of items 1-8, wherein the product further comprises an excipient or carrier, e.g., at least one of a sustained release agent, a filler, a binder, a humectant, a disintegrant, an absorption enhancer, a surfactant, and a lubricant.

[0045] 10. The use of any one of items 1-9, wherein the product is: a human food, a human pharmaceutical, an animal food, or an animal pharmaceutical, or a kit.

[0046] 11. The use of any one of items 1-10, wherein the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the foregoing are capable of inducing adipocyte apoptosis.

[0047] 12. A method of inducing adipocyte apoptosis in a sample or subject, comprising adding to the sample or subject a vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the foregoing, wherein optionally an effective amount of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the foregoing is at least 1 mg (for humans) or at least 0.1 mg (e.g., for cats, dogs) or at least 0.015 mg (e.g., for mice, rats).

[0048] 13. A product for preventing and / or alleviating and / or treating obesity, overweight, or a symptom or a state or a tendency or a disease or a disorder caused by or associated with overweight or obesity, or for weight loss in a mammal, comprising an effective amount of vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist, and analogs of the above, and an excipient or a carrier; wherein optionally the effective amount is at least 1 mg (for humans) or at least 0.1 mg (for example for cats, dogs) or at least 0.015 mg (for example for mice, rats).

[0049] 14. The product of item 13, wherein the vitamin D and analogs thereof include but are not limited to vitamin D2, vitamin D3, vitamin D4, (R)-vitamin D2, 11-hydroxymeridinol, 1a-hydroxy vitamin D4; preferably vitamin D3.

[0050] The 25-hydroxyvitamin D and analogs thereof include but are not limited to 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 25,26-dihydroxyvitamin D3, 5,6-trans vitamin D2, 25-hydroxyvitamin D3 monohydrate.

[0051] The vitamin D receptor agonist includes but is not limited to 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D2, trans calcitriol.

[0052] 15. The product of item 13, which is a food for humans, a drug for humans, a food for animals, or a drug for animals, or a kit comprising the food for humans, the drug for humans, the food for animals, or the drug for animals.

[0053] 16. The product of any one of items 13-15, wherein the mammal includes a human, a dog, a cat, a mouse, a rat.

[0054] 17. The product of any one of items 13-16, wherein the excipient or the carrier is at least one of a sustained release agent, a filler, a binder, a humectant, a disintegrant, an absorption enhancer, a surfactant, and a lubricant.

[0055] 18. The product of any one of items 13-17, wherein the symptom or the state or the tendency or the disease or the disorder caused by or associated with overweight or obesity includes but is not limited to diabetes, fatty liver, hypertension.

[0056] 19. A method of preventing and / or alleviating or treating obesity, overweight, or a symptom or a state or a tendency or a disease or a condition caused by or associated with overweight or obesity in a mammal, comprising administering to a subject in need of preventing and / or alleviating or treating obesity, overweight, or a symptom or a state or a tendency or a disease or a condition caused by or associated with overweight or obesity in a mammal, an effective amount of vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist, and analogs of the above.

[0057] 20. The method of item 19, wherein the effective amount of vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist, and analogs of the above is at least 1 mg (for humans) or at least 0.1 mg (e.g., for cats, dogs) or at least 0.015 mg (e.g., for mice, rats).

[0058] 21. The method of item 19 or 20, wherein the vitamin D and analogs thereof include, but are not limited to, vitamin D2, vitamin D3, vitamin D4, (R)-vitamin D2, 11-hydroxymeridinol, 1a-hydroxyvitamin D4; preferably vitamin D3.

[0059] the 25-hydroxyvitamin D and analogs thereof include, but are not limited to, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 25,26-dihydroxyvitamin D3, 5,6-trans vitamin D2, 25-hydroxyvitamin D3 monohydrate.

[0060] the vitamin D receptor agonist includes, but is not limited to, 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D2, trans calcitriol.

[0061] 22. The method of item 21, wherein the vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist, and analogs of the above are in the form of a food for humans, a drug for humans, a food for animals, or a drug for animals.

[0062] 23. The method of item 19, wherein the subject includes: a human, a dog, a cat, a mouse, a rat.

[0063] 24. The method of item 19, wherein the symptom or a state or a tendency or a disease or a condition caused by or associated with overweight or obesity includes, but is not limited to, diabetes, fatty liver, hypertension.

[0064] 25. The method of any one of items 19-24, wherein the vitamin D, 25-hydroxyvitamin D, or a vitamin D receptor agonist, and analogs of the above are in a form suitable for intravenous injection, intraperitoneal injection, subcutaneous injection, or oral administration.

[0065] 26. The method of any one of items 19-25, wherein the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the foregoing are administered at a frequency of once, twice, three times or seven times per week, for a period of 7 days or more, e.g., 7-60 days.

[0066] The term

[0067] The vitamin D is a fat-soluble vitamin that plays an important role in regulating calcium and phosphorus levels in the body and mineralized bone, which includes, among others: vitamin D2 (also known as ergocalciferol, CAS No. 50-14-6); vitamin D3 (also known as cholecalciferol; colecalciferol, CAS No. 67-97-0); vitamin D4 (also known as 22-dihydroergocalciferol, CAS No. 511-28-4); vitamin D3 (also known as cholecalciferol; colecalciferol, CAS No. 67-97-0).

[0068] The analogs of the vitamin D include 11-hydroxysugiol (CAS No. 88664-08-8); calcitetrol (1a, 24, 25-trihydroxy VD3) (CAS No. 72203-93-1); 1a-hydroxy vitamin D4 (CAS No. 143032-85-3); previtamin D3 (CAS No. 1173-13-h); 7-dehydrocholesterol (CAS No. 434-16-2); calcitriol impurities D (CAS No. 103656-40-2); (R)-vitamin D2 (CAS No. 116559-84-3); (24S)-24, 25-dihydroxyvitamin D3 ((24S)-24, 25-dihydroxycholecalciferol) (CAS No. 55700-58-8); 22-hydroxyvitamin D3 (CAS No. 110927-46-3).

[0069] Vitamin D can act directly after entering the body or also via metabolism to 25-hydroxyvitamin D or further hydroxylated to 1,25-dihydroxyvitamin D, which is used to activate the vitamin D receptor in target tissues (also called VDR receptor agonist).

[0070] The 25-hydroxyvitamin D is the main circulating form of vitamin D and mainly includes: 25-hydroxyvitamin D2(also known as Ercalcidiol, CAS No. 21343-40-8); 25-hydroxyvitamin D3(also known as Calcifediol or Calcitriol, CAS No. 19356-17-3); 25, 26-dihydroxyvitamin D3(also known as 25, 26-dihydroxycholecalciferol or 25, 26-dihydroxycholecalciferol, CAS No. 29261-12-9); 25-hydroxyvitamin D4.

[0071] The analogs of 25-hydroxyvitamin D include: 5,6-trans-Vitamin D2 (51744-66-2); Calcifediol monohydrate (also known as 25-hydroxyvitamin D3 monohydrate, or 25-hydroxy Vitamin D3 monohydrate, CAS Number 63283-36-3); Ercalcitriol (1α,25-Dihydroxy Vitamin D2) (CAS Number 60133-18-8); Secalciferol ((24R)-24,25-Dihydroxyvitamin D3) (CAS Number 55721-11-4); 1α,25-Dihydroxyprevitamin D3 (CAS Number 57102-09-7); 1α-Hydroxy-3-epi-vitamin D3 (CAS Number 58028-00-5); (1S)-Calcitriol (1α,25-Dihydroxy-3-epi-vitamin-D3) (CAS Number 61476-45-7); 5,6-trans-Vitamin D3 (5,6-trans-Cholecalciferol; 5,6-trans-Colecalciferol) (CAS Number 22350-41-0); Vitamin D3 octanoate (CAS Number 927822-16-0); 24,25-DihydroxyVD3 (CAS Number 40013-87-4); Doxercalciferol (CAS Number 54573-75-0).

[0072] The vitamin D receptor agonist refers to any compound that directly binds to and activates the vitamin D receptor, which can be naturally occurring, such as 1,25-dihydroxy vitamin D2, 1,25-dihydroxy vitamin D3; or synthetic, such as, 24,25-dihydroxy vitamin D2(24,25-Dihydroxy VD2, CAS No. 58050-55-8); trans-Doxercalciferol (CAS No. 74007-20-8); Alfacalcidol (1-hydroxycholecalciferol) (CAS No. 41294-56-8); Inecalcitol (TX 522) (CAS No. 163217-09-2); Lumisterol 3 (9β,10α-Cholesta-5,7-dien-3β-ol) (CAS No. 5226-01-7); Calcitriol Derivatives (CAS No. 2070009-24-2); MC 976 (CAS No. 129831-99-8); Atocalcitol (CAS No. 302904-82-1); (24R)-MC 976 (CAS No. 112828-09-8); Pyrocholecalciferol (CAS No. 10346-43-7); Dihydrotachysterol (CAS No. 67-96-9); Lexacalcitol (KH 1060) (CAS No. 131875-08-6); Isotachysterol 3 (CAS No. 22350-43-2); Pefcalcitol (SMD-502) (CAS No. 381212-03-9); Maxacalcitol (22-Oxacalcitriol) (CAS No. 103909-75-7); 2MD (CAS No. 213250-70-5); Eldecalcitol (ED-71) (CAS No. 104121-92-8); CB 1151 (CAS No. 182369-28-4); VDR agonist 1 (compound 28) (CAS No. 2269494-43-9).

[0073] The terms “reduce body weight”, “weight loss” and “weight reduction” can be used interchangeably.

[0074] "Body mass index" or "BMI" means the ratio of weight (kg) divided by height (m) squared.

[0075] "Overweight" is defined (e.g., for adults) as having a BMI of 25 to 30 (according to WHO's criteria) (individuals with this BMI are at risk of obesity).

[0076] "Obesity" is defined (e.g., for adults) as having a BMI of greater than or equal to 30 (according to WHO's criteria), e.g., greater than or equal to 35 (greater than or equal to 35 to less than 40 is called class 2 obesity), e.g., greater than or equal to 40 (or called class 3 obesity), e.g., greater than or equal to about 45, 50, 55, or 60.

[0077] In Asian countries, "overweight" means a body mass index of greater than or equal to 23 to less than 25 (individuals with this BMI are at risk of obesity); "obesity" means a body mass index of greater than or equal to 25, e.g., greater than or equal to 30, 45, 50, 55, or 60.

[0078] The term "overweight" as used herein is meant to include all of the above definitions of overweight.

[0079] The term "obesity" as used herein is a state in which the natural energy reserves stored in the adipose tissue of animals, particularly humans and other mammals, are increased to a point that is associated with certain health conditions or increased mortality. The term "obesity" as described herein is meant to include all of the above definitions of obesity, including but not limited to obesity, abdominal obesity, visceral obesity, central obesity, or abdominal adiposity.

[0080] The term "disease or condition caused by or associated with overweight or obesity" as described herein includes diabetes, fatty liver, hypertension, cardiovascular disease, atherosclerosis, dyslipidemia, nonalcoholic steatohepatitis, pulmonary hypertension, infertility, cancer, e.g., breast cancer, colorectal cancer, hepatocellular carcinoma.

[0081] The term "excipient or carrier" as used herein means any acceptable material, compound or excipient currently on the market, such as a liquid solid or filler, a stabilizer, a dispersant, a suspending agent, a diluent, a binder, a lubricant, a disintegrant, a thickening agent, a colorant, a flavoring agent, a preservative, a solvent or an encapsulating material, for example, for transporting or transporting the compound for the invention to the subject, so that it can perform its desired function.

[0082] Administration / dosage / formulation

[0083] The compounds of the application or the products of the application can be prepared in a conventional carrier or excipient, according to the form of administration appropriate for the user, for example in a form known in the art suitable for oral, vaginal, parenteral, nasal, intravenous, subcutaneous, intestinal or any other suitable mode of administration, containing an organic or inorganic carrier. If necessary, the products of the application can also be used in combination with other active agents, for example other anorectic agents. The products of the application can also be used in combination with other methods of weight loss, for example physical exercise.

[0084] Suitable dosage forms of the products of the application include, for example, tablets, capsules, pills, dispersions, suspensions, solutions, syrups, granules, pellets, patches, gels, powders, suppositories, injections, drops, liquid sprays for nasal or oral administration, dry powders or spray formulations for inhalation, and the like.

[0085] Methods of administration of the products of the application include, but are not limited to, oral administration, topical administration, rectal administration, parenteral, intravenous administration, subcutaneous administration, inhalation, intranasal administration.

[0086] For oral use, tablets, dragees, liquids, drops, suppositories, or capsules, caplets and soft capsules are particularly suitable. The products of the application intended for oral use can be prepared according to any method known in the art and can include one or more selected from the group consisting of inert, non-toxic pharmaceutically acceptable carriers suitable for manufacturing said dosage forms.

[0087] The products for use in the present application can be delivered directly to the targeted cells / tissues / organs, for example abdominal tissues.

[0088] In the present application, the administration dose of the vitamin D, 25-hydroxy vitamin D, vitamin D receptor agonist, and analogs of the above substances to a non-human mammal is 0.1-100 mg / kg (dose / body weight) (excluding 0.1 mg / kg), for example, 0.1-100 mg / kg, 0.1-90 mg / kg, 0.1-80 mg / kg, 0.1-60 mg / kg, 0.1-50 mg / kg, 0.1-40 mg / kg, 0.1-30 mg / kg, 0.1-20 mg / kg, 0.1-10 mg / kg, 0.1-6 mg / kg, 0.1-4 mg / kg, 0.1-2 mg / kg, 0.2-100 mg / kg, 0.2-90 mg / kg, 0.2-80 mg / kg, 0.2-70 mg / kg, 0.2-60 mg / kg, 0.2-50 mg / kg, 0.2-40 mg / kg, 0.2-30 mg / kg, 0.2-20 mg / kg, 0.2-10 mg / kg, 0.2-8 mg / kg, 0.2-6 mg / kg, 0.2-4 mg / kg, 0.3-100 mg / kg, 0.3-80 mg / kg, 0.3-60 mg / kg, 0.3-40 mg / kg, 0.3-20 mg / kg, 0.5-100 mg / kg, 0.5-90 mg / kg, 0.5-80 mg / kg, 0.5-70 mg / kg, 0.5-60 mg / kg, 0.5-50 mg / kg, 0.5-40 mg / kg, 0.5-30 mg / kg, 0.5-20 mg / kg, 0.5-10 mg / kg, 0.6-90 mg / kg, 0.6-80 mg / kg, 0.6-70 mg / kg, 0.6-60 mg / kg, 0.6-50 mg / kg, 0.6-40 mg / kg, 0.6-20 mg / kg, 0.6-10 mg / kg, 0.6-7 mg / kg, 0.8-100 mg / kg, 0.8-90 mg / kg, 0.8-80 mg / kg, 0.8-60 mg / kg, 1-100 mg / kg, 1-80 mg / kg, 1-70 mg / kg, 1-60 mg / kg, 1-50 mg / kg, 1-40 mg / kg, 1-30 mg / kg, 1-20 mg / kg, 1-10 mg / kg, 2-80 mg / kg, 2-60 mg / kg, 2-40 mg / kg, 5-100 mg / kg, 5-80 mg / kg, 5-60 mg / kg, 5-40 mg / kg, 6-80 mg / kg, 6-60 mg / kg, 6-40 mg / kg, 6-20 mg / kg, 6-10 mg / kg, 8-80 mg / kg, 8-60 mg / kg, 8-40 mg / kg, 8-20 mg / kg, 10-80 mg / kg, 10-60 mg / kg, 10-40 mg / kg, 15-80 mg / kg, 15-60 mg / kg, 15-40 mg / kg, 20-80 mg / kg, 20-60 mg / kg, 20-40 mg / kg, 30-80 mg / kg, 30-50 mg / kg, 40-80 mg / kg, 40-60 mg / kg, 40-50 mg / kg.

[0089] kit

[0090] In another aspect of the application, an article of manufacture or kit useful for treating or preventing obesity is provided. The article of manufacture or kit comprises one or more containers and a label or package insert on or associated with the container(s). Suitable containers include, for example, bottles, vials, syringes, IV solution bags, SC solution injectors, etc. The containers can be formed from a variety of materials such as glass or plastic. The container holds a composition itself, or a composition in combination with another agent effective for treating or preventing the obesity, and can have a sterile access port (for example the container can be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is a vitamin D, 25-hydroxyvitamin D, vitamin D receptor agonist, and analogs of the above described. The label, package insert, or such file indicates that the composition is used for treating obesity.

[0091] In certain embodiments, the composition contained in the container of the article of manufacture or kit is formulated for subcutaneous injection or intravenous injection. The article of manufacture or kit in this embodiment of the application can further comprise a product insert indicating the mode of administration of the compound, for example by intravenous injection or subcutaneous injection or oral administration, preferably subcutaneous injection or oral administration, wherein the frequency of administration can be 1, 2, 3 or 7 times per week, for a period of 7 days or more, for example 7-60 days, optionally the product insert indicates that the compound is to be administered at a dose of at least 1 mg (for humans) or at least 0.1 mg (for example for cats, dogs) or at least 0.015 mg (for example for mice, rats) per week; preferably the product insert indicates that when used on humans and the weekly dose reaches 6 mg or more, careful use is required and the period should not exceed two weeks.

[0092] In addition, the article of manufacture or kit can further include pharmaceutically-acceptable buffers, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It can further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes. BRIEF DESCRIPTION OF DRAWINGS

[0093] Figure 1 The gradual increase in body weight of mice during high-fat diet induction is shown; the horizontal axis of the graph represents time, and the vertical axis represents the body weight of mice, red (HFD) is a high-fat diet mouse, and black (SD) is a normal diet mouse. (n = 10, bar: ± SEM)

[0094] Figure 2 The body weight of high-fat diet-induced mice is shown compared with normal diet mice; the vertical axis of the graph represents the body weight of mice, gray is a high-fat diet mouse, and black is a normal diet mouse, Before indicates before high-fat diet, and After indicates after 12 weeks of high-fat diet. (n = 10, bar: ± SEM, ****: p < 0.0001, ns: p ≥ 0.05)

[0095] Figure 3 The body fat rate of high-fat diet-induced mice is shown compared with normal diet mice; the vertical axis of the graph represents the body fat rate of mice, gray is a high-fat diet mouse, and black is a normal diet mouse. (n = 10, bar: ± SEM, ****: p < 0.0001)

[0096] Figure 4 The design of the small molecule vitamin D3 weight loss experiment is shown;

[0097] Figure 5The figure shows the significant decrease in body weight of mice during the treatment with vitamin D3: the horizontal axis of the figure represents time, the vertical axis represents the body weight of mice, red (HFD+VD3) is the mice treated with vitamin D3 under high-fat diet conditions, and black (HFD+DMSO) is the mice treated with the same volume of DMSO solution under high-fat diet conditions. (n=10, bar: ±SEM);

[0098] Figure 6 The figure shows the comparison of body weight of mice before and after treatment with vitamin D3: the vertical axis of the figure represents the body weight of mice, dark gray is the mice treated with vitamin D3 under high-fat diet conditions, light gray is the mice treated with the same volume of DMSO solution under high-fat diet conditions, and black is the mice treated with the same volume of DMSO solution under normal diet conditions, Before represents before high-fat diet, and After represents after 12 weeks of high-fat diet. (n=10, bar: ±SEM, ****: p<0.0001, ns: p≥0.05);

[0099] Figure 7 The figure shows the comparison of body fat content of mice before and after treatment with vitamin D3: the vertical axis of the figure represents the body fat content of mice, dark gray is the mice treated with vitamin D3 under high-fat diet conditions, light gray is the mice treated with the same volume of DMSO solution under high-fat diet conditions, and black is the mice treated with the same volume of DMSO solution under normal diet conditions, Before represents before high-fat diet, and After represents after 12 weeks of high-fat diet. (n=10, bar: ±SEM, ****: p<0.0001, ns: p≥0.05);

[0100] Figure 8 The figure shows the comparison of lean body mass of mice before and after treatment with vitamin D3: the vertical axis of the figure represents the lean body mass of mice, dark gray is the mice treated with vitamin D3 under high-fat diet conditions, light gray is the mice treated with the same volume of DMSO solution under high-fat diet conditions, and black is the mice treated with the same volume of DMSO solution under normal diet conditions, Before represents before high-fat diet, and After represents after 12 weeks of high-fat diet. (n=10, bar: ±SEM);

[0101] Figure 9 The figure shows the comparison of average daily food intake per body weight of each mouse: the vertical axis of the figure represents the average daily food intake per body weight of each mouse, gray is the mice treated with vitamin D3 under high-fat diet conditions, and black is the mice treated with the same volume of DMSO solution under high-fat diet conditions. (n=10, bar: ±SEM, ns: p≥0.05);

[0102] Figure 10Results of the mice rotarod test (assessing motor balance ability) after vitamin D3 treatment are shown: the vertical axis in the figure represents the average time recorded in the rotarod test for the last three times, • for mice treated with an equal volume of DMSO solution under normal diet conditions, ■ for mice treated with an equal volume of DMSO solution under high-fat diet conditions, and ▲ for mice treated with vitamin D3 under high-fat diet conditions. (n = 10, bar: ± SEM, **: p < 0.01, ns: p ≥ 0.05);

[0103] Figure 11 Results of the mice endurance test after vitamin D3 treatment are shown: the vertical axis in the figure represents the total distance run on the treadmill during the mice endurance test, • for mice treated with an equal volume of DMSO solution under normal diet conditions, ■ for mice treated with an equal volume of DMSO solution under high-fat diet conditions, and ▲ for mice treated with vitamin D3 under high-fat diet conditions. (n = 10, bar: ± SEM, *: p < 0.05);

[0104] Figure 12 Results of the mice glucose tolerance test (assessing the function of blood glucose regulation) after vitamin D3 treatment are shown: the horizontal axis in the figure represents time, and the vertical axis represents the blood glucose concentration of the mice, red (HFD+VD3) for mice treated with vitamin D3 under high-fat diet conditions, and black (HFD+DMSO) for mice treated with an equal volume of DMSO solution under high-fat diet conditions. (n = 10, bar: ± SEM, **: p < 0.01, *: p < 0.05);

[0105] Figure 13 Comparison of HE staining images of mouse adipose tissue is shown: the figure is an HE staining image of paraffin sections of mouse inguinal adipose tissue; among them, the left column represents mice treated with an equal volume of DMSO solution under normal diet conditions, the middle column represents mice treated with an equal volume of DMSO solution under high-fat diet conditions, and the right column represents mice treated with vitamin D3 under high-fat diet conditions. (n = 2, scale bar: 200 μm);

[0106] Figure 14 Design of the mice adipose tissue apoptosis detection experiment is shown: the mice were first treated with a high-fat diet or a normal diet for 12 weeks, and then treated with vitamin D3 or an equal volume of DMSO solution for 2 weeks (the diet conditions of the mice during this period remained the same as before), and then TUNEL detection was performed;

[0107] Figure 15Statistical analysis of the percentage of TUNEL-positive cells in mouse adipose tissue: the vertical axis of the graph represents the percentage of TUNEL-positive cells (%); SD represents a normal diet, HFD represents a high-fat diet, black is a mouse treated with an equal volume of DMSO solution under a normal diet, light gray is a mouse treated with an equal volume of DMSO solution under a high-fat diet, and dark gray is a mouse treated with a high dose of vitamin D3 under a high-fat diet. (n = 5, error bar: standard error, *: p < 0.05)

[0108] Figure 16 Long-term experimental design of small molecule vitamin D3 is shown; Vitamin D3 is used to treat for four weeks while a high-fat diet is used to induce obesity, and then a high-fat diet is maintained while the effect is evaluated;

[0109] Figure 17 Long-term effect after vitamin D3 treatment is shown: the horizontal axis of the graph represents time, and the vertical axis represents the body weight of mice. Red (HFD+VD3) is a mouse treated with vitamin D3 for the first four weeks under a high-fat diet, and black (HFD+DMSO) is a mouse treated with an equal volume of DMSO solution for the first four weeks under a high-fat diet. Among them, treatment represents the four-week period of treatment with vitamin D3 or an equal volume of DMSO solution, and withdrawal represents the nine-week period after the treatment with vitamin D3 or an equal volume of DMSO solution is stopped. (n = 13-15, bar: ± SEM, ****: p < 0.0001, ***: p < 0.001);

[0110] Figure 18 Body fat content of mice 90 days after the end of vitamin D3 treatment is shown: the vertical axis of the graph represents the body fat content of mice, and gray is a mouse treated with vitamin D3 for four weeks under a high-fat diet and then stopped for three months, and black is a mouse treated with an equal volume of DMSO solution for four weeks under a high-fat diet and then stopped for three months. During the period when the treatment with vitamin D3 or an equal volume of DMSO solution is stopped, the mice are always maintained on a high-fat diet. (n = 10, bar: ± SEM, *: p < 0.05);

[0111] Figure 19 Survival curve of mice at the end of vitamin D3 treatment is shown: red (Vitamin D3) is the survival curve of mice treated with vitamin D3 for four weeks under a high-fat diet, and black (DMSO) is the survival curve of mice treated with an equal volume of DMSO solution for four weeks under a high-fat diet. The mice are always maintained on a high-fat diet (n = 15);

[0112] Figure 20Comparison of HE staining of mouse liver: HE staining of paraffin sections of mouse liver tissue, left panel: mice treated with vitamin D3 for 4 weeks and then stopped for 3 months under high-fat diet, right panel: mice treated with equal volume of DMSO solution for 4 weeks and then stopped for 3 months under high-fat diet. The right panel shows a large number of fat particles in the liver tissue, which is a typical symptom of fatty liver (n = 5, scale: 200 μm (top), 100 μm (bottom));

[0113] Figure 21 Comparison of the proportion of fat particle area in the same size field after HE staining of mouse liver tissue: the vertical axis in the figure represents the proportion of fat particle area in the same size field. (n = 5, bar: ± SEM, ****: p < 0.0001);

[0114] Figure 22 Design of experiment of prevention of obesity in mice by small molecule vitamin D3;

[0115] Figure 23 Prevention of weight gain in mice by vitamin D3 under high-fat diet: the horizontal axis in the figure represents time, and the vertical axis represents the weight of mice, red (VD3) for mice treated with vitamin D3, black (Control) for mice treated with equal volume of DMSO solution, all mice were given high-fat diet. (n = 10, bar: ± SEM);

[0116] Figure 24 Prevention of increase of body fat in mice by vitamin D3 under high-fat diet: the vertical axis in the figure represents the content of body fat in mice, gray for mice treated with vitamin D3 under high-fat diet, black for mice treated with equal volume of DMSO solution under high-fat diet. (n = 10, bar: ± SEM, ****: p < 0.0001);

[0117] Figure 25 Prevention of decline of exercise function in mice by vitamin D3 under high-fat diet: the vertical axis in the figure represents the average time recorded in the last three times in the rotarod experiment of mice, gray for mice treated with vitamin D3 under high-fat diet, black for mice treated with equal volume of DMSO solution under high-fat diet. (n = 10, bar: ± SEM, *: p < 0.05);

[0118] Figure 26 Prevention of decline of blood glucose regulation function in mice by vitamin D3 under high-fat diet: the horizontal axis in the figure represents time, and the vertical axis represents the concentration of blood glucose in mice, red (VD3) for mice treated with vitamin D3 under high-fat diet, black (Control) for mice treated with equal volume of DMSO solution under high-fat diet. (n = 10, bar: ± SEM, *: p < 0.05);

[0119] Figure 27 Figure 8 shows the relative content of vitamin D3 in different tissues of mice. The horizontal axis represents different tissues, and the vertical axis represents the relative content of vitamin D3. Untreated represents untreated, and Treated represents 72 hours after the third injection. Black represents untreated mice, and gray represents treated mice. (n = 5, error bar: standard error, ****: p < 0.0001, **: p < 0.01)

[0120] Figure 28 Figure 9 shows the results of TUNEL detection of paraffin sections of adipose tissue of mice. Blue represents DAPI, green represents TUNEL, and blue-green represents colocalization of DAPI and TUNEL. Row 1 shows the results of DAPI staining, row 2 shows the results of TUNEL staining in the same field, and row 3 shows the results of colocalization of DAPI and TUNEL in the same field. Column 1 represents mice treated with an equal volume of DMSO under normal dietary conditions, column 2 represents mice treated with an equal volume of DMSO under high-fat dietary conditions, and column 3 represents mice treated with a high dose of vitamin D3 under high-fat dietary conditions. (n = 5, scale bar: 50 μm). DETAILED DESCRIPTION

[0121] In order to make the objectives, technical solutions, and advantages of the present application clearer, the present application is further described in detail below with reference to specific examples and with reference to the accompanying drawings.

[0122] In the following examples, the methods used are conventional methods unless otherwise specified, and the reagents used are commercially available reagents unless otherwise specified.

[0123] MATERIALS AND METHODS

[0124] 1. Experimental mice

[0125] The experimental mice used in this study were all from Beijing Vital River Laboratory Animal Technology Co., Ltd., and were all C57BL / 6N female mice with an initial age of 8 weeks.

[0126] 2. Experimental mouse feeding

[0127] All experimental mice in this study were raised under specific pathogen free (SPF) conditions, using an IVC system, with 12 hours of light and 12 hours of darkness alternating, and the temperature was controlled between 22°C and 26°C. The bedding was poplar shavings, and the mouse maintenance feed was replaced at least once a week. Newly purchased experimental mice needed to be acclimated in this feeding environment for one week before the experiment.

[0128] All experimental mice involved in this study enjoyed the due animal welfare, and all animal experiments in this study were in line with the animal ethics.

[0129] 3. High-fat diet induction

[0130] The high-fat feed used in all experiments in this study for high-fat diet induction was sourced from Research Diets' primary agent Beijing Kaiguokong Technology Co., Ltd., which was a 60kcal% high-fat feed.

[0131] During the high-fat diet induction period, the mouse cage must be placed with a chew stick, and the high-fat feed must be supplemented at least once every 5 days.

[0132] 4. Intraperitoneal injection

[0133] The steps for the intraperitoneal injection operation of experimental mice are as follows:

[0134] 1) Shake the solution to be injected well;

[0135] 2) Use a 1mL syringe to draw the solution, and expel the excess air in the syringe;

[0136] 3) Place the syringe on the experimental bench;

[0137] 4) Grasp and fix the experimental mouse in the correct way;

[0138] 5) Turn the experimental mouse's abdomen upward and its head slightly downward with the left hand slightly turned outward, so that its internal organs move to the upper abdomen;

[0139] 6) Hold the syringe with the right hand, and insert the needle into the skin about 1cm from the right side of the experimental mouse's lower abdomen, then insert it into the abdominal cavity at about 45 degrees toward the abdominal midline;

[0140] 7) Push the syringe with the right thumb, inject a certain volume of solution according to the experimental requirements, and ensure that the line of sight is always parallel to the scale of the syringe;

[0141] 8) Pull out the syringe and put the experimental mouse back into the cage.

[0142] 5. Body composition detection

[0143] The instrument used for body composition detection is the small animal body composition analyzer of Echo MRI International Medical Equipment Co., Ltd., and the operation is based on the instruction manual.

[0144] 6. Rotarod test

[0145] The instrument used for the rotarod test in this study is the mouse rotarod fatigue instrument of Jiangsu Saiyan Technology Co., Ltd., with a shaft diameter of 60mm.

[0146] The experimental procedure was set as follows: the rotating speed of the rotating rod was accelerated from 4 r / min to 44 r / min in 300 s, the acceleration was 8 r / min2, and the total duration of the experimental procedure was 5 min.

[0147] 7. Tolerance experiment

[0148] The instrument used in the tolerance experiment was a small animal treadmill from Jiangsu Saiyuan Biotechnology Co., Ltd., the length of the runway was 60 cm, and the inclination (slope) of the treadmill was set to 15 degrees.

[0149] 8. Glucose tolerance test

[0150] The instrument used in the glucose tolerance test was a blood glucose meter from Roche, and the test paper was Roche blood glucose test paper,

[0151] 9. RNA extraction

[0152] The RNA extraction was performed using a magnetic bead method blood, cell, and tissue total RNA extraction kit from Tiangen Biotech (Beijing) Co., Ltd., and the instrument used was a full-automatic nucleic acid extraction and purification instrument from the same company. The specific experimental process was divided into sample pretreatment and full-automatic nucleic acid extraction and purification instrument operation.

[0153] The specific steps of sample pretreatment are as follows:

[0154] 1) Take out fresh or -80℃ frozen tissues of appropriate size;

[0155] 2) Add a certain amount of liquid nitrogen and grind the tissues into powder;

[0156] 3) Add 500 μL TRIzol reagent, and then transfer them together to a 1.5 mL centrifuge tube;

[0157] 4) Mix well with a pipette, and stand for 2 min;

[0158] 5) Centrifuge at 12000 rpm for 3 min;

[0159] 6) Absorb 450 μL supernatant (if it is adipose tissue, it will be divided into three layers after centrifugation, first absorb the upper liquid, and then absorb the remaining supernatant) into another 1.5 mL centrifuge tube;

[0160] 7) Add 20 μL proteinase K, and mix well with a pipette.

[0161] The specific steps of using the full-automatic nucleic acid extraction instrument are as follows:

[0162] 1) Take out the 96-deep-well plate from the kit;

[0163] 2) Mix the 96-deep-well plate several times upside down;

[0164] 3) Remove vacuum packaging, centrifuge at 500 rpm for 30 s;

[0165] 4) Tear off the sealing film;

[0166] 5) Add the tissue supernatant containing protease K to the first column and the seventh column of the 96 deep-well plate;

[0167] 6) Add 10 μL of Dnase I stock solution and 70 μL of Buffer RDD to the third column and the ninth column of the 96 deep-well plate;

[0168] 7) Turn on the automatic nucleic acid extractor;

[0169] 8) Place the 96 deep-well plate on the 96 deep-well plate base of the automatic nucleic acid extraction and purification instrument;

[0170] 9) Insert the matching magnetic rod sleeve into the corresponding card slot of the automatic nucleic acid extraction and purification instrument;

[0171] 10) Select the corresponding first program and click to run;

[0172] 11) After the first program is run, take out the 96 deep-well plate;

[0173] 12) Add 700 μL of deproteinization solution RD to the third column and the ninth column of the 96 deep-well plate;

[0174] 13) Place the 96 deep-well plate again on the corresponding base of the automatic nucleic acid extraction and purification instrument;

[0175] 14) Select the corresponding second program and click to run;

[0176] 15) After the automatic extraction program is completed, take out the 96 deep-well plate;

[0177] 16) Absorb the RNA solution in the fifth column and the eleventh column of the 96 deep-well plate and store it at -80°C;

[0178] 17) Take out the matching magnetic rod sleeve and discard it;

[0179] 18) After all the samples are extracted, turn off the automatic nucleic acid extraction and purification instrument.

[0180] 10. Preparation of mouse adipose tissue paraffin sections

[0181] In the process of preparing mouse adipose tissue paraffin sections, the main instruments and equipment used are SAKURA paraffin tissue embedding machine and Leica paraffin section machine

[0182] 11. HE staining of paraffin sections

[0183] The specific steps of HE staining of paraffin sections of each tissue and organ of mice are as follows:

[0184] 1) Place the paraffin section in xylene for 15 min;

[0185] 2) Take the paraffin section out of xylene and place it in another xylene for 10 min;

[0186] 3) Take the paraffin section out of xylene and place it in the third xylene for 5 min;

[0187] 4) Take the paraffin section out of xylene and place it in a mixture of 50% xylene + 50% ethanol for 10 min;

[0188] 5) Take the paraffin section out of the mixture of 50% xylene + 50% ethanol and place it in 100% ethanol for 10 min;

[0189] 6) Take the paraffin section out of anhydrous ethanol and place it in another anhydrous ethanol for 5 min;

[0190] 7) Take the paraffin section out of anhydrous ethanol and place it in the third anhydrous ethanol for 2 min;

[0191] 8) Take the paraffin section out of anhydrous ethanol and place it in 95% ethanol solution for 5 min;

[0192] 9) Take the paraffin section out of 95% ethanol solution and rinse with running water for 3 min;

[0193] 10) Place the paraffin section in hematoxylin staining solution for 10 min;

[0194] 11) Rinse with running water for 5 min;

[0195] 12) Place the paraffin section in 1% hydrochloric acid ethanol solution for 10 s;

[0196] 13) Rinse with running water for 10 min;

[0197] 14) Place the paraffin section under a microscope for observation (ensure that the paraffin section is in a wet state), and if the staining is insufficient, return to step 10) for re-staining

[0198] 15) Rinse with running water for 5 min;

[0199] 16) Place the paraffin section in eosin staining solution for 4 min;

[0200] 17) Take the paraffin section out of eosin staining solution and place it in 80% ethanol solution for 10 s;

[0201] 18) Remove the paraffin sections from the 80% ethanol solution and place them in 95% ethanol solution for 1 min; 19) Remove the paraffin sections from the 95% ethanol solution and place them in anhydrous ethanol for 2 min;

[0202] 20) Observe the paraffin sections under a microscope (ensure the paraffin sections are moist).

[0203] If the staining is insufficient, proceed backwards to step 17) and re-stain.

[0204] 21) Place the paraffin sections in anhydrous ethanol for 5 minutes;

[0205] 22) Remove the paraffin sections from the anhydrous ethanol and place them in a mixture of 50% xylene and 50% ethanol for 10 minutes;

[0206] 23) Remove the paraffin sections from the mixture of 50% xylene and 50% paraffin and place them in xylene for 10 minutes;

[0207] 24) Remove the paraffin sections from the xylene and place them in another xylene solution for 10 minutes;

[0208] 25) Remove the paraffin section from the xylene, drop a drop of neutral resin on it, and cover it with a coverslip to seal the slide;

[0209] 26) Place the sealed paraffin sections in a ventilated place overnight.

[0210] 12. TUNEL assay of paraffin sections in mouse adipose tissue: The specific steps for TUNEL assay of paraffin sections in mouse adipose tissue are as follows:

[0211] 1) Place the paraffin sections in xylene for 15 minutes;

[0212] 2) Remove the paraffin sections from the xylene and place them in another xylene solution for 10 minutes;

[0213] 3) Remove the paraffin sections from the xylene and place them in a third xylene solution for 5 minutes;

[0214] 4) Remove the paraffin sections from the xylene and place them in a mixture of 50% xylene and 50% ethanol for 10 minutes;

[0215] 5) Remove the paraffin sections from the mixture of 50% xylene and 50% ethanol and place them in 100% ethanol for 10 minutes;

[0216] 6) Remove the paraffin sections from the anhydrous ethanol and place them in another anhydrous ethanol solution for 5 minutes;

[0217] 7) Remove the paraffin sections from the anhydrous ethanol and place them in a third batch of anhydrous ethanol for 2 minutes;

[0218] 8) The paraffin section is taken out of the absolute ethanol and placed in a 95% ethanol solution for 5 minutes;

[0219] 9) The paraffin section is taken out of the 95% ethanol solution and rinsed with running water for 3 minutes;

[0220] 10) 20 μg / ml of proteinase K without DNase is added dropwise (diluted with 10 mM Tris-HCl with a pH of 7.4 to 7.8) and left at room temperature for 15 to 30 minutes;

[0221] 11) Rinsed 3 times with PBS buffer;

[0222] 12) The TUNEL detection solution is prepared and added dropwise to the tissue sample and left at 37°C in the dark for 60 minutes;

[0223] 13) Rinsed 3 times with PBS buffer;

[0224] 14) DAPI staining solution is added dropwise and left at room temperature in the dark for 15 minutes;

[0225] 15) Rinsed 3 times with PBS buffer;

[0226] 16) Placed under a fluorescence microscope for observation.

[0227] 13. Pretreatment for ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) detection

[0228] The specific steps of the pretreatment of the sample before detection using the method of UPLC-MS detection of the relative content of vitamin D3 in each tissue organ of mice are as follows:

[0229] 1) The mouse is first dissected to obtain the required tissue organs;

[0230] 2) 20 mg (± 0.5 mg) of the tissue sample is accurately weighed using an analytical balance and placed in a 1.5 mL centrifuge tube;

[0231] 3) The 1.5 mL centrifuge tube is placed on ice and the tissue sample in the 1.5 mL centrifuge tube is completely cut into a paste using FST spring scissors;

[0232] 4) 200 μL of methanol (HPLC grade) containing a known concentration of vitamin D3 standard is added in advance and the residual tissue sample on the FST spring scissors is ensured to be in the methanol;

[0233] 5) The 1.5 mL centrifuge tube containing the tissue sample and methanol is placed in a high-power ultrasonic cell disrupter and extracted at 4°C for 20 minutes;

[0234] 6) 18000rpm 4°C centrifugation for 30min;

[0235] 7) Absorb the supernatant (as clean as possible) and transfer to another 1.5 mL centrifuge tube;

[0236] 8) 18000rpm 4°C centrifugation for 30min;

[0237] Absorb the supernatant and transfer to another 1.5 mL centrifuge tube for UPLC-MS detection.

[0238] 14. Statistical analysis of experimental data

[0239] The experimental data in this study were statistically analyzed using Excel, GraphPad, R language, etc. Image J was used to process all fluorescence staining images and chemical staining images. For all two-group experimental data, the t-test method was used to analyze the significance of the difference; for all multi-group experimental data, the ANOVA test method was used to analyze the significance of the difference. Among them, ****: p<0.0001, ***: p<0.001, **: p<0.01, *: p<0.05, ns: p≥0.05.

[0240] Example 1 High-fat diet-induced mouse obesity

[0241] In this study, obese mice were used as research objects, so before the study, first of all, we need to obtain obese mice that meet the experimental conditions. In this study, 2-month-old wild-type female C57BL / 6N mice were fed with 60kcal% high-fat diet, thereby obtaining an obese mouse model induced by high-fat diet.

[0242] 1.1 High-fat diet-induced mouse weight gain

[0243] After 12 weeks of high-fat diet feeding, the average weight of the mice was initially 20.2g, and after 4 weeks, it reached 25.9g, after 8 weeks, it reached 31.0g, and finally, after 12 weeks, it gradually increased to 38.3g; while the average weight of the mice without high-fat diet was initially 20.2g, and after 12 weeks of normal diet, the average weight slowly increased to 25.2g Figure 1 )。

[0244] To evaluate the effect of high-fat diet on the body weight of mice more intuitively, the body weight of mice before and after 12 weeks of high-fat diet induction was compared with that of mice of the same batch on a normal diet by T-test. The results showed that there was no significant difference in the body weight of the two groups of mice before high-fat diet induction; while the body weight of mice after 12 weeks of high-fat diet induction was significantly higher than that of mice on a normal diet, and the difference was extremely significant. Therefore, compared with normal diet, high-fat diet can significantly increase the body weight of mice, which is consistent with the expected modeling effect. Figure 2 ).

[0245] 1.2 High-fat diet induces an increase in the body fat rate of mice

[0246] To further confirm the effect of high-fat diet on inducing obesity in mice, the body weight of mice after 12 weeks of high-fat diet induction and that of mice of the same batch on a normal diet were detected. The results showed that the average body fat rate of mice after high-fat diet induction was 37.62%, while the average body fat rate of mice on a normal diet was 14.32%, and the degree of obesity of mice on a high-fat diet was much higher than that of mice on a normal diet; and statistical analysis showed that the difference between the two was extremely significant. Figure 3 ).

[0247] Therefore, by feeding mice with high-fat feed for a period of time, an obese mouse model that meets the conditions can be obtained; the obese mouse model used in the subsequent experiments of this study was obtained by this method.

[0248] Example 2: Small molecule vitamin D3 achieves weight loss in obese mice

[0249] To determine the weight loss effect of small molecule vitamin D3, a batch of experiments was designed. First, obese mice were obtained by 12 weeks of high-fat diet induction, then small molecule vitamin D3 was administered to obese mice after high-fat diet by intraperitoneal injection (the diet of mice remained unchanged during the entire experiment), and then detection was performed (the experimental process is shown in Figure 4 ). The single dose of small molecule vitamin D3 was 2 mg / kg body weight, and the administration method was intraperitoneal injection, 3 times a week (injection once on Monday, Wednesday, and Friday each week), and the administration period was 4 weeks. In addition, this study also set up a control group, in which the same batch of obese mice induced by high-fat diet were injected with the same volume of DMSO solution, and the same batch of mice on a normal diet were also injected with the same volume of DMSO solution.

[0250] The detection indexes include: body weight, body fat, body fat rate (body fat / body weight), lean body mass, food intake / body weight, rotarod test (to evaluate the motor balance ability), endurance test (to detect the endurance using a treadmill / running machine), glucose tolerance test (to detect the blood glucose regulation function), histological detection (to observe the staining after preparing paraffin sections of mouse tissues). Note: Except for body weight, body fat, body fat rate, lean body mass, food intake / body weight, other detection indexes are each independently used for a batch of mice for experiments.

[0251] 2.1 Small molecule vitamin D3 reduces the body weight of obese mice

[0252] In this study, the body weight of mice was measured during the 4-week treatment of small molecule vitamin D3. The results showed that, compared with the obese mice injected with the same volume of DMSO solution in the same batch, the obese mice treated with small molecule vitamin D3 for 4 weeks, even on a high-fat diet throughout, still showed a significant decrease in body weight. Specifically, the average body weight of obese mice treated with small molecule vitamin D3 for 4 weeks decreased from 39.3g before treatment to 25.7g; while the average body weight of obese mice injected with the same volume of DMSO solution was 38.3g before treatment and 38.8g after treatment. Figure 5

[0253] In order to more intuitively evaluate the effect of small molecule vitamin D3 treatment on reducing the body weight of obese mice, this study used statistical methods to perform T-test on the body weight of obese mice treated with small molecule vitamin D3 for 4 weeks, the body weight of obese mice injected with the same volume of DMSO solution in the same batch, and the body weight of non-obese mice injected with the same volume of DMSO solution on a normal diet.

[0254] The statistical results showed that, compared with the obese mice injected with the same volume of DMSO solution in the same batch, the obese mice treated with small molecule vitamin D3 for 4 weeks had a very significant difference in body weight; while compared with the non-obese mice injected with the same volume of DMSO solution on a normal diet in the same batch, the body weight of the two groups had no significant difference. Figure 6 Therefore, small molecule vitamin D3 treatment can significantly reduce the body weight of obese mice within 4 weeks, making it comparable to normal diet mice.

[0255] 2.2 Small molecule vitamin D3 reduces the body fat of obese mice

[0256] In order to further confirm the weight loss effect of small molecule vitamin D3, this study also conducted body composition detection on obese mice treated with small molecule vitamin D3 for 4 weeks, obese mice injected with the same volume of DMSO solution in the same batch, and non-obese mice injected with the same volume of DMSO solution on a normal diet. ​

[0257] The results of the detection show that the average body fat content of the obese mice treated with the small molecule vitamin D3 for 4 weeks is reduced from 16.0 g to 6.2 g, and the body fat content is reduced by 61.5%; the average body fat content of the obese mice injected with the same volume of DMSO solution in the same batch is 15.4 g, the average body fat content of the non-obese mice on a normal diet injected with the same volume of DMSO solution in the same batch is 4.3 g; and through statistical analysis, the average body fat content of the obese mice treated with the small molecule vitamin D3 has a very significant difference from the body fat content of the obese mice injected with the same volume of DMSO solution, but has no significant difference from the average body fat content of the non-obese mice on a normal diet injected with the same volume of DMSO solution. Figure 7 ) It can be seen that the small molecule vitamin D3 can greatly reduce the body fat content of the obese mice, and the weight loss effect is very significant.

[0258] In addition, the present study evaluated its effect on the lean body mass of mice by detecting the body composition of mice. The results show that after being treated with the small molecule vitamin D3 for 4 weeks, the lean body mass of the obese mice appears to be slightly reduced, and the average lean body mass is reduced from 19.0 g to 16.9 g Figure 8 ).

[0259] 2.3 The food intake of mice does not significantly decrease during the treatment with the small molecule vitamin D3

[0260] Among the small molecules or drugs that have been reported to have a weight loss effect, some of them reduce food intake by reducing appetite, thereby achieving weight loss. In order to confirm whether the small molecule vitamin D3 has an effect on the food intake of mice while achieving weight loss, the food intake of the obese mice on a high-fat diet was measured regularly during the treatment with the small molecule vitamin D3 or the same volume of DMSO solution.

[0261] The results show that the ratio of the average daily food intake per mouse to the body weight of the obese mice treated with the small molecule vitamin D3 does not significantly decrease compared to the obese mice treated with the same volume of DMSO solution in the same batch Figure 9 ). It can be seen that the small molecule vitamin D3 does not significantly reduce the food intake of obese mice during the treatment, and the weight loss effect is not achieved by reducing the food intake of obese mice.

[0262] 2.4 The small molecule vitamin D3 improves the exercise function of obese mice

[0263] Obesity leads to decreased motor function. To better assess the impact of small-molecule vitamin D3 on motor function while promoting weight loss, this study designed two batches of experiments. Obese mice were induced to become obese through a high-fat diet for 12 consecutive weeks. These obese mice were then treated with small-molecule vitamin D3 via intraperitoneal injection (dietary conditions remained constant throughout the experiment). The single dose of small-molecule vitamin D3 was 2 mg / kg body weight, administered intraperitoneally three times a week (once on Monday, Wednesday, and Friday) for four weeks. Control groups were set up for both batches of experiments. Mice from the same batch of high-fat diet-induced obese mice received an equal volume of DMSO solution, as did mice from the same batch on a normal diet. The study assessed the balance and endurance of the obese mice treated with small-molecule vitamin D3 in both batches to confirm whether small-molecule vitamin D3 could improve motor function in mice while promoting weight loss.

[0264] Small molecule vitamin D3 enhances motor balance in obese mice

[0265] To assess the changes in motor balance in obese mice treated with small molecule vitamin D3, this study conducted rotarod experiments on a batch of obese mice treated with small molecule vitamin D3 for 4 weeks, obese mice injected with an equal volume of DMSO solution, and non-obese mice on a normal diet injected with an equal volume of DMSO solution to detect their motor balance.

[0266] The rotarod experiment results showed that, after being injected with the same volume of DMSO solution, non-obese mice on a normal diet had an average effective running time of 178.1 s on the rotarod, while obese mice induced by a high-fat diet had an average effective running time of only 130.3 s, proving that obesity can indeed significantly reduce motor balance. Furthermore, after 4 weeks of treatment with small-molecule vitamin D3, the obese mice had an average effective running time of 179.6 s on the rotarod, which was essentially the same as that of non-obese mice on a normal diet injected with the same volume of DMSO solution. Figure 10 ).

[0267] Statistical analysis showed that the rotarod test results of obese mice treated with small-molecule vitamin D3 were significantly different from those of obese mice injected with the same volume of DMSO solution in the same batch; however, there was no significant difference compared with non-obese mice on a normal diet that were injected with the same volume of DMSO solution. Figure 10 This demonstrates that small-molecule vitamin D3, while achieving weight loss, can also significantly improve the motor balance ability of mice that have declined due to obesity, restoring it to the level of non-obese mice with a normal diet.

[0268] Small molecule vitamin D3 enhances endurance in obese mice

[0269] To assess the changes in endurance in obese mice treated with small molecule vitamin D3, this study conducted a treadmill test on another batch of obese mice treated with small molecule vitamin D3 for 4 weeks, obese mice injected with an equal volume of DMSO solution from the same batch, and non-obese mice on a normal diet injected with an equal volume of DMSO solution to test their endurance.

[0270] The treadmill experiment results showed that, after receiving the same volume of DMSO solution, non-obese mice on a normal diet ran an average distance of 557.5 m on a 15-degree incline treadmill, while obese mice induced by a high-fat diet ran an average distance of only 274.0 m, proving that obesity can indeed significantly reduce endurance. Furthermore, after 4 weeks of treatment with small-molecule vitamin D3, obese mice ran an average distance of 361.3 m on a 15-degree incline treadmill, a significant improvement compared to obese mice injected with the same volume of DMSO solution in the same batch. Figure 11 ).

[0271] Statistical analysis showed that the endurance test results of obese mice treated with small-molecule vitamin D3 were significantly different from those of obese mice injected with the same volume of DMSO solution in the same batch. Figure 11 This shows that while small-molecule vitamin D3 can help mice lose weight, it can also significantly improve the endurance of mice that have lost weight due to obesity, but there is still a large gap between the endurance level of mice with normal diets and that of non-obese mice.

[0272] 2.5 Small molecule vitamin D3 improves glycemic regulation in obese mice

[0273] Obesity can impair an individual's ability to regulate blood glucose and may further lead to diabetes. To assess the impact of small-molecule vitamin D3 on blood glucose regulation while promoting weight loss, this study designed a new batch of experiments. First, obese mice were induced to become obese through a high-fat diet, fed a high-fat diet for 12 consecutive weeks. Then, these obese mice were treated with small-molecule vitamin D3 via intraperitoneal injection (the mice's dietary conditions remained unchanged throughout the experiment). The single dose of small-molecule vitamin D3 was 2 mg / kg body weight, injected three times a week (Monday, Wednesday, and Friday) for four weeks. In addition, a control group was included, which received an equal volume of DMSO solution from the same batch of obese mice induced by the high-fat diet. Glucose tolerance tests were performed on these obese mice treated with small-molecule vitamin D3 for four weeks and on the obese mice injected with the same volume of DMSO solution to assess their blood glucose regulation.

[0274] The results of the glucose tolerance test showed that the fasting blood glucose concentration of the obese mice treated with the small molecule vitamin D3 for 4 weeks was not significantly different from that of the obese mice injected with the same volume of DMSO solution in the same batch, but the blood glucose concentrations at 15 min, 30 min, 45 min, 60 min, and 90 min were significantly lower. Through statistical analysis, the blood glucose concentrations of the obese mice treated with the small molecule vitamin D3 at 30 min, 60 min, and 90 min were significantly different from those of the obese mice injected with the same volume of DMSO solution in the same batch, and the blood glucose concentrations at 45 min were very significantly different. Figure 12 Thus, the small molecule vitamin D3 can improve blood glucose regulation while achieving weight loss.

[0275] 2.6 Small molecule vitamin D3 does not significantly change fat cell size while achieving weight loss

[0276] Among the small molecules or drugs reported to have a weight loss effect, whether by inhibiting fat intake or increasing fat consumption, fundamentally, they all achieve weight loss by reducing the size of fat cells. To explore the mechanism of weight loss of the small molecule vitamin D3, this study dissected obese mice treated with the small molecule vitamin D3 for 4 weeks, obese mice injected with the same volume of DMSO solution in the same batch, and non-obese mice injected with the same volume of DMSO solution, and then took the inguinal fat tissue to make paraffin sections, which were then stained with HE and observed and photographed under a microscope to compare the size of fat cells under different conditions and preliminarily explore the mechanism of weight loss of the small molecule vitamin D3.

[0277] By observing the HE-stained paraffin sections of fat tissue under different conditions, this study found that the size of fat cells in obese mice treated with the small molecule vitamin D3 for 4 weeks was not significantly different from that of obese mice injected with the same volume of DMSO solution in the same batch, and both were significantly larger than that of non-obese mice injected with the same volume of DMSO solution. Figure 13 As previously proven, the small molecule vitamin D3 can significantly reduce the body fat content of obese mice, and the size of fat cells in obese mice treated with the small molecule vitamin D3 for 4 weeks is similar to that of obese mice injected with the same volume of DMSO solution in the same batch, indicating that the number of fat cells in mice treated with the small molecule vitamin D3 has decreased. Thus, the mechanism of the small molecule vitamin D3 in reducing body fat and achieving weight loss is likely to be achieved by reducing the number of fat cells.

[0278] 2.7 Vitamin D3 treatment can induce adipocyte apoptosis

[0279] The inventors further investigated the reason for the decrease of adipocytes, and analyzed the ratio of TUNEL positive cells in adipocytes (see experimental design flow chart in Figure 14 ).

[0280] The results are shown in Figure 15 and Figure 28 , since TUNEL positivity reflects to some extent the occurrence of apoptosis, this result indicates that the decrease of adipocytes in mice after vitamin D3 treatment can be due to adipocyte apoptosis.

[0281] In addition, the inventors also observed that the livers of mice on high fat diet were significantly enlarged (fatty liver symptoms), while the livers of mice treated with vitamin D3 were significantly reduced (but larger than normal diet mice), and the symptoms were significantly improved, indicating that vitamin D3 has a certain therapeutic effect on fatty liver induced by obesity.

[0282] In summary, vitamin D3 treatment can significantly reduce the body fat percentage of mice, while improving exercise balance and endurance, improving blood glucose regulation, and improving fatty liver symptoms; and can reduce the number of adipocytes.

[0283] 2.8 Dosage of vitamin D3

[0284] In the above examples of the present application, a large dose of vitamin D3, such as 2 mg / kg, is used. The inventors further investigated the dosage and found that only a large dose (e.g. greater than 0.1 mg / kg, 1 mg / kg or more, such as 0.1-10 mg / kg (not including 0.1 mg / kg), 1-10 mg / kg) can have the effect described.

[0285] Example 3 Vitamin D3 treatment has a long-term weight loss effect

[0286] Many small molecules or drugs reported to have a weight loss effect have a certain weight loss effect, but after stopping the small molecule treatment or drug delivery, the body weight and body fat content are prone to rebound.

[0287] In order to evaluate whether the small molecule vitamin D3 has a long-term effect, a new set of experiments was designed, in which the mice were continuously fed with high-fat feed by inducing obesity through high-fat diet; at the same time, the mice on high-fat diet were treated with small molecule vitamin D3 by intraperitoneal injection, and then detected (see experimental flow chart in Figure 16). The single dose of the small molecule vitamin D3 is 2 mg / kg body weight, and the administration method is intraperitoneal injection, 3 times a week (1st, 3rd, 5th day every 7 days, i.e. once a week on Monday, Wednesday and Friday), and the administration starting time is synchronized with the starting time of the high-fat diet; the administration cycle is 4 weeks. The control group is set in this study, and the mice of the same batch subjected to the high-fat diet are injected with the same volume of DMSO solution. It is emphasized that, regardless of the experimental group or the control group, the mice are always maintained on the high-fat diet during the treatment of the small molecule vitamin D3 or the same volume of DMSO solution and after the treatment is stopped, and the effect of the small molecule vitamin D3 is evaluated under this condition.

[0288] The detection indexes include: body weight, body fat, body fat rate (body fat / body weight), histological detection (after preparing paraffin sections of mouse tissues, staining is observed). Note: the histological detection is performed by using a separate batch of mice for the experiment.

[0289] The vitamin D3 treatment can maintain the body weight stable for a long time

[0290] In this study, by tracking the body weight of the mice for a long time, it is found that, under the condition of always maintaining the high-fat diet, the average body weight of the mice in the control group increases from 19.5 g at the beginning to 23.7 g after 4 weeks, and to 34.2 g after 9 weeks after the injection is stopped; regardless of the 4-week period of injecting the same volume of DMSO solution or the 9-week period after the injection is stopped, the body weight of the mice is in a continuous increasing process. The average body weight of the mice in the experimental group is 18.9 g at the beginning; during the treatment of the small molecule vitamin D3, the body weight of the mice decreases rapidly in the first two weeks, and the average body weight decreases to 15.4 g after two weeks; the body weight gradually stabilizes in the last two weeks, and the average body weight of the mice is stabilized at about 15.0 g after the treatment of the small molecule vitamin D3 for 4 weeks under the condition of the high-fat diet. Figure 17

[0291] After the treatment of the small molecule vitamin D3 is stopped, the body weight of the mice increases very slowly from the second week, and the body weight increase rate is significantly lower than that of the control group. Figure 17

[0292] After the treatment of the small molecule vitamin D3 is stopped for 9 weeks, the average body weight of the mice in the experimental group finally increases slowly to 26.0 g, which is much lower than the average body weight of the mice in the control group. Figure 17

[0293] Through statistical analysis, after the injection of the small molecule vitamin D3 or the same volume of DMSO solution is stopped, although the mice are always maintained on the high-fat diet, there is a very significant difference between the measured body weight data of the mice in the experimental group and the control group at each time point. Figure 17 ​​​). Thus, even after a short period of treatment with micromolar vitamin D3, the mice were able to maintain a lower body weight for a longer period of time, even while being maintained on a high fat diet, demonstrating that micromolar vitamin D3 has long term effects. The experimental mice were treated with micromolar vitamin D3 for only 4 weeks, with a total of 12 intraperitoneal injections, and yet the experimental mice were able to maintain a lower body weight for 9 weeks after treatment was stopped, and the rate of weight gain was slower than the control mice starting at 2 weeks after treatment was stopped.

[0294] Vitamin D3 treatment can maintain low body fat for a long period of time

[0295] To further confirm the long term effects of micromolar vitamin D3, the inventors measured the body composition of the mice 90 days after treatment with micromolar vitamin D3 was stopped, to assess the level of body fat. The results showed that the average body fat content of the control mice was 13.4 g after 90 days of injection with an equal volume of DMSO, while the average body fat content of the experimental mice was only 7.0 g after 90 days of treatment with micromolar vitamin D3, which was much lower than the control mice. Statistical analysis showed that there was a significant difference between the two groups (p < 0.05). Figure 18 Thus, even after a short period of treatment with micromolar vitamin D3, the mice were able to maintain a lower body fat content for a longer period of time, even while being maintained on a high fat diet, further demonstrating that micromolar vitamin D3 has long term effects.

[0296] Vitamin D3 treatment can increase the survival rate of mice on a high fat diet

[0297] The inventors further studied the survival rate of the mice after 4 weeks of treatment with vitamin D3, and the results are shown in Figure 19 which showed that the survival rate of the mice treated with vitamin D3 was always higher than the control mice after the mice died while being maintained on a high fat diet. Thus, vitamin D3 not only has long term weight loss effects, but also can prolong the lifespan of mice on a high fat diet.

[0298] Vitamin D3 treatment greatly improves fatty liver symptoms

[0299] See Figure 20 where the left panel shows a mouse that was treated with vitamin D3 for 4 weeks and then stopped treatment for 3 months while being maintained on a high fat diet, and the right panel shows a mouse that was treated with an equal volume of DMSO for 4 weeks and then stopped treatment for 3 months while being maintained on a high fat diet. The right panel shows a large number of fat particles in the liver tissue, which is a typical symptom of fatty liver; while the left panel shows no obvious fat particles, indicating that the mouse that was treated with vitamin D3 and then continued to be maintained on a high fat diet for 3 months had significantly improved fatty liver symptoms.

[0300] Vitamin D3 treatment greatly improved the symptoms of fatty liver (quantitative statistics)

[0301] See Figure 21 The results show that the mice treated with vitamin D3 continue to have a very significant improvement in the symptoms of fatty liver (or basically no symptoms of fatty liver) after three months of high-fat diet compared with the control group.

[0302] In addition, during the dissection of the mice, the heart, spleen, lung, kidney, small intestine, colon, brain, skin, and muscle (gastrocnemius) were also taken for paraffin section and HE staining. None of these organs showed obvious abnormalities in both the experimental and control groups. In addition, before dissection, fasting blood glucose was also detected, and the fasting blood glucose of the mice treated with vitamin D3 was significantly lower than that of the control group (the fasting blood glucose of the control group was significantly higher, which was a symptom of diabetes).

[0303] In summary, it is shown that after treatment with vitamin D3, the mice have a long-term weight loss effect under the condition of continuous high-fat diet, the symptoms of fatty liver are greatly improved, and the fasting blood glucose is closer to the normal level. In addition, through long-term observation, the survival rate of mice treated with vitamin D3 under the condition of continuous high-fat diet is higher than that of the control group.

[0304] Example 4: Small molecule vitamin D3 can prevent obesity in mice

[0305] It has been proven that small molecule vitamin D3 has a significant weight loss effect. In order to confirm whether small molecule vitamin D3 can prevent obesity, a new batch of experiments was designed. First, 2-month-old mice were fed with high-fat feed (60kcal% high-fat feed) to induce obesity. At the same time, the mice on high-fat diet were treated with small molecule vitamin D3 by intraperitoneal injection. The single dose of small molecule vitamin D3 was 1mg / kg body weight, the administration method was intraperitoneal injection, the administration frequency was 3 times a week (1st, 3rd, and 5th day every 7 days, i.e. Monday, Wednesday, and Friday every week), the administration starting time was synchronized with the starting time of high-fat diet, and the administration cycle was 12 weeks. In addition, a control group was set up, and the same batch of mice on high-fat diet were injected with the same volume of DMSO solution, and the two were compared to evaluate the preventive effect of small molecule vitamin D3 on obesity (see Figure 22 ).

[0306] Detection indicators: body weight, body fat, rotarod test (to evaluate the motor balance ability), glucose tolerance test (to detect the blood glucose regulation function). Note: Except for body weight and body fat, other detection indicators were tested using a separate batch of mice for each detection indicator.

[0307] Small molecule vitamin D3 can prevent the increase of body weight of mice on high-fat diet

[0308] In this study, the body weight of mice was measured during the 12-week process of high-fat diet and small molecule vitamin D3 treatment. The results showed that under the condition of continuous high-fat diet, the body weight of mice injected with the same volume of DMSO solution continued to increase; while the body weight of mice treated with small molecule vitamin D3 remained stable during the whole process of high-fat diet, the average body weight before the experiment was 20.1 g, and the average body weight after 12 weeks was 23.4 g, without significant increase. Figure 23 Therefore, small molecule vitamin D3 can effectively prevent the increase of body weight of mice on high-fat diet.

[0309] Small molecule vitamin D3 can prevent the increase of body fat of mice on high-fat diet

[0310] In order to further confirm the effect of small molecule vitamin D3 on preventing obesity, this study also detected the body composition of mice treated with 12-week high-fat diet and small molecule vitamin D3, and the same batch of mice treated with 12-week high-fat diet and injection of the same volume of DMSO solution.

[0311] The results showed that under the condition of continuous high-fat diet, the average body fat content of mice injected with the same volume of DMSO solution reached 14.4 g after 12 weeks; while the average body fat content of mice treated with small molecule vitamin D3 was only 3.8 g after 12 weeks, which was significantly lower than that of the control group; through statistical analysis, there was a very significant difference between the two groups. Figure 24 Therefore, under the condition of continuous high-fat diet, treatment with small molecule vitamin D3 can effectively prevent the increase of body fat of mice.

[0312] Small molecule vitamin D3 can prevent the decline of motor function of mice on high-fat diet

[0313] Obesity can lead to a decline in exercise capacity, in order to better assess the impact of small molecule vitamin D3 on exercise function while preventing obesity, this study designed a new batch of experiments, first by high-fat diet induced obesity, the mouse was fed with high-fat diet; At the same time, through the intraperitoneal injection of the drug to the mouse with high-fat diet with small molecule vitamin D3 treatment. Single dose of small molecule vitamin D3 is 1mg / kg body weight, each injection on Monday, Wednesday, Friday, the treatment period is 12 weeks. This study also sets up a control group, the same batch of mice with high-fat diet were injected with the same volume of DMSO solution. This study through to the same time high-fat diet and small molecule vitamin D3 treatment of mice, and the same batch of mice with high-fat diet and injection of the same volume of DMSO solution were tested by the rotarod test to evaluate their exercise function.

[0314] The results of the rotarod test showed that under the condition of continuous high-fat diet, the average effective time of the mice injected with the same volume of DMSO solution on the rotating shaft after 12 weeks was 128.0s; While the small molecule vitamin D3 treated mice even with high-fat diet, the average effective time of running on the rotating shaft after 12 weeks was 159.1s, which was significantly better than the control group; Through statistical analysis, there was a significant difference between the two( Figure 25 ). Therefore, under the condition of continuous high-fat diet, the mice treated with small molecule vitamin D3 have stronger exercise balance ability; It is proved that the treatment with small molecule vitamin D3 can prevent the decline of exercise function caused by obesity in mice to a certain extent.

[0315] Small molecule vitamin D3 can prevent the decline of blood glucose regulation function in high-fat diet mice

[0316] In order to evaluate the impact of small molecule vitamin D3 on blood glucose regulation function while preventing obesity, this study designed a batch of experiments, first by high-fat diet induced obesity, the mouse was fed with high-fat diet; At the same time, through the intraperitoneal injection of the drug to the mouse with high-fat diet with small molecule vitamin D3 treatment. Single dose of small molecule vitamin D3 is 1mg / kg body weight, each injection on Monday, Wednesday, Friday, the treatment period is 12 weeks. This study also sets up a control group, the same batch of mice with high-fat diet were injected with the same volume of DMSO solution. This study through to the same time high-fat diet and small molecule vitamin D3 treatment of mice, and the same batch of mice with high-fat diet and injection of the same volume of DMSO solution were tested by the rotarod test to evaluate their exercise function.

[0317] The results of the glucose tolerance test showed that, under the condition of continuous high-fat diet, the fasting blood glucose concentration of the mice treated with the small molecule vitamin D3 did not have obvious difference compared with the mice of the same batch injected with the same volume of DMSO solution, and the blood glucose concentrations at 5 time points of 15 min, 30 min, 45 min, 60 min and 90 min were significantly reduced. Through statistical analysis, the blood glucose concentrations of the mice treated with the small molecule vitamin D3 at 4 time points of 30 min, 45 min, 60 min and 90 min had significant difference compared with the mice of the same batch injected with the same volume of DMSO solution. Figure 26 ) It can be seen that, under the condition of continuous high-fat diet, the mice treated with the small molecule vitamin D3 can not only effectively prevent obesity, but also can well prevent the decrease of blood glucose regulation function.

[0318] The above shows that, after treatment with vitamin D3 (for example, at a dose of 1 mg / kg), the mice can prevent the increase of body weight and body fat, improve the decrease of exercise function caused by obesity, and prevent the symptoms of diabetes under the condition of continuous high-fat diet.

[0319] Example 5 Enrichment of vitamin D3 in the adipose tissue of mice after treatment

[0320] A 2-month-old wild-type female C57BL / 6N mouse was treated by intraperitoneal injection with vitamin D3 at a dose of 2 mg / kg, and the injection was performed every two days. The mouse was dissected 72 hours after the third injection, and 11 kinds of tissue organs including heart, liver, spleen, lung, kidney, brain, small intestine, colon, muscle (gastrocnemius), blood and adipose tissue were obtained. The relative content of vitamin D3 in these tissue organs was detected by ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS).

[0321] Through the statistics of the relative content of vitamin D3 in the 11 kinds of tissue organs of the untreated and the mouse 72 hours after the third injection, it was found that the content of vitamin D3 in the adipose tissue of the mouse 72 hours after the third injection was much higher than that of the untreated mouse, and the difference was very significant. Among the remaining 10 kinds of tissue organs, except that the content of vitamin D3 in the liver was almost the same before and after the third injection, vitamin D3 could not be detected in the brain whether before treatment or 72 hours after the third injection, and the content of vitamin D3 in the other tissue organs after the third injection was higher than that of the untreated mouse. However, for the mouse 72 hours after the third injection, the content of vitamin D3 in the 10 kinds of tissue organs was much lower than that in the adipose tissue, and the difference was extremely significant.

[0322] The results are shown in the following table:Figure 27 The results show that vitamin D3 can be enriched in fat tissue, and the content is much higher than that in other tissue organs. Vitamin D3 reduces the number of fat cells to achieve the effect of weight loss because a large amount of vitamin D3 is enriched in fat tissue after high-dose vitamin D3 treatment, and the content is much higher than that in other tissue organs, thereby leading to fat cell apoptosis.

[0323] The above specific embodiments further specifically describe the purposes, technical solutions and beneficial effects of the present application. It should be understood that the above description is only a specific embodiment of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. Use of vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above in the manufacture of a product, wherein the product is for preventing and / or alleviating or treating obesity, overweight, or a symptom or a state or a tendency or a disease or a disorder caused by or associated with overweight or obesity, or for reducing body weight in a mammal.

2. Use according to claim 1, wherein, The effective amount of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above is at least 1 mg (for human) or at least 0.1 mg (for example for cat, dog) or at least 0.015 mg (for example for mouse, rat).

3. Use according to claim 1 or 2, wherein, The vitamin D and analogs thereof include but are not limited to vitamin D2, vitamin D3, vitamin D4, (R)-vitamin D2, 11-hydroxymeridinol, 1α-hydroxy vitamin D4; preferably vitamin D3; The 25-hydroxyvitamin D and analogs thereof include but are not limited to 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 25,26-dihydroxyvitamin D3, 5,6-trans vitamin D2, 25-hydroxyvitamin D3 monohydrate; The vitamin D receptor agonist includes but is not limited to 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D2, trans maxacalcitol.

4. The use according to any one of claims 1 to 3, wherein, The vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above are in a form suitable for intravenous injection, intraperitoneal injection, subcutaneous injection or oral administration.

5. The use according to any one of claims 1 to 4, wherein, The administration dose of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above in a non-human mammal is 0.1-100 mg / kg.

6. The use according to any one of claims 1 to 5, wherein, The product further comprises a product instruction indicating that the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above are administered at a dose of 0.1-100 mg / kg, or indicating that the effective amount of the vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above is at least 1 mg (for human) or at least 0.1 mg (for example for cat, dog) or at least 0.015 mg (for example for mouse, rat), optionally the product instruction indicates that the administration frequency is 1 time, 2 times, 3 times or 7 times per week, and the administration cycle is more than 7 days, for example 7-60 days.

7. The use according to any one of claims 1 to 6, wherein, The mammal includes: human, dog, cat, mouse, rat.

8. The use according to any one of claims 1 to 7, wherein, The symptom or state or tendency or disease or disorder caused by or associated with overweight or obesity includes but is not limited to: diabetes, fatty liver, hypertension.

9. The use according to any one of claims 1 to 8, wherein, The product further comprises an excipient or carrier, for example at least one of a sustained release agent, a filler, a binder, a humectant, a disintegrant, an absorption promoter, a surfactant and a lubricant.

10. Use according to any one of claims 1 to 9, wherein, The product is: a food for human, a drug for human, a food for animal or a drug for animal, or a kit.

11. Use according to any one of claims 1 to 10, wherein, The vitamin D, 25-hydroxyvitamin D or vitamin D receptor agonist and analogs of the above can induce apoptosis of adipocytes.

12. A method of inducing apoptosis of adipocytes in a sample or a subject, comprising adding to the sample or subject vitamin D, 25-hydroxyvitamin D or a vitamin D receptor agonist and analogs of the foregoing, wherein optionally the effective amount of the vitamin D, 25-hydroxyvitamin D or a vitamin D receptor agonist and analogs of the foregoing is at least 1 mg (for humans) or at least 0.1 mg (for example for cats, dogs) or at least 0.015 mg (for example for mice, rats).

13. A product for preventing and / or alleviating and / or treating obesity, overweight, or a symptom or a state or a tendency or a disease or a disorder caused by or associated with overweight or obesity, or for weight loss in a mammal, comprising an effective amount of vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist and analogs of the foregoing, and an excipient or a carrier; wherein optionally the effective amount is at least 1 mg (for humans) or at least 0.1 mg (for example for cats, dogs) or at least 0.015 mg (for example for mice, rats).

14. The product of claim 13, wherein, The vitamin D and analogs thereof include, but are not limited to, vitamin D2, vitamin D3, vitamin D4, (R)-vitamin D2, 11-hydroxymeridinol, 1α-hydroxy vitamin D4; preferably vitamin D3; The 25-hydroxyvitamin D and analogs thereof include, but are not limited to, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 25,26-dihydroxyvitamin D3, 5,6-trans vitamin D2, 25-hydroxyvitamin D3 monohydrate; The vitamin D receptor agonist includes, but is not limited to, 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D2, trans calcitriol.

15. The product of claim 13, which is a human food, a human drug, an animal food or an animal drug, or a kit comprising the human food, the human drug, the animal food or the animal drug.

16. The product of any one of claims 13-15, wherein the mammal comprises: The human, the dog, the cat, the mouse, the rat.

17. The product of any one of claims 13-16, wherein, The excipient or the carrier is at least one of a sustained release agent, a filler, a binder, a humectant, a disintegrant, an absorption enhancer, a surfactant and a lubricant.

18. The product of any one of claims 13-17, wherein, The symptom or a state or a tendency or a disease or a disorder caused by or associated with overweight or obesity includes, but is not limited to, diabetes, fatty liver, hypertension.

19. A method of preventing and / or alleviating or treating obesity, overweight, or a symptom or a state or a tendency or a disease or a disorder caused by or associated with overweight or obesity in a mammal, comprising administering to a subject in need of preventing and / or alleviating or treating obesity, overweight, or a symptom or a state or a tendency or a disease or a disorder caused by or associated with overweight or obesity in a mammal an effective amount of vitamin D, 25-hydroxyvitamin D, a vitamin D receptor agonist and analogs of the foregoing.

20. The method of claim 19, wherein the effective amount of vitamin D, 25-hydroxyvitamin D, vitamin D receptor agonist, and analogs of the foregoing is at least 1 mg (for humans) or at least 0.1 mg (e.g., for cats, dogs) or at least 0.015 mg (e.g., for mice, rats).

21. The method of claim 19 or 20, wherein, The vitamin D and analogs thereof include, but are not limited to, vitamin D2, vitamin D3, vitamin D4, (R)-vitamin D2, 11-hydroxytocols, 1 α-hydroxy vitamin D4; preferably vitamin D3; The 25-hydroxyvitamin D and analogs thereof include, but are not limited to, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 25,26-dihydroxyvitamin D3, 5,6-trans vitamin D2, 25-hydroxyvitamin D3 monohydrate; The vitamin D receptor agonist includes, but is not limited to, 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D2, trans maxacalcitol.

22. The method of claim 21, wherein, The vitamin D, 25-hydroxyvitamin D, vitamin D receptor agonist, and analogs of the foregoing are in a form of a human food, a human drug, an animal food, or an animal drug.

23. The method of claim 19, wherein the subject comprises: The human, dog, cat, mouse, rat.

24. The method of claim 19, wherein, The condition or state or tendency or disease or disorder caused by or associated with overweight or obesity includes, but is not limited to, diabetes, fatty liver, hypertension.

25. The method of any one of claims 19-24, wherein, The vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist, and analogs of the foregoing are in a form suitable for intravenous injection, intraperitoneal injection, subcutaneous injection, or oral administration.

26. The method of any one of claims 19-25, wherein the vitamin D, 25-hydroxyvitamin D, or vitamin D receptor agonist, and analogs of the foregoing are administered at a frequency of 1, 2, 3, or 7 times per week, for a period of 7 days or more, e.g., 7-60 days.

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