TWI432203B - Use of lanostane and poria extract in treating cachexia - Google Patents

Description

Use of lanosterol and anthraquinone extract for the treatment of cachexia

The present invention relates to a pharmaceutical composition for preventing and treating a wasting disease, such as cachexia and anorexia, especially a cachexia caused by cancer. The pharmaceutical composition of the present invention contains a lanosterol compound as an active ingredient. A suitable source of this lanosterol compound is the hydrazine extract. In addition, Cordyceps sinensis, Antrodia camphorata and Ganoderma lucidum are also sources of possible lanosterol compounds.

Cachexia indicates that it is in a bad state of weakness, often caused by diseases causing anorexia and endocrine and immune system changes, fatigue, muscle and visceral protein reduction, and ultimately weight loss.

Cachexia is a common symptom in chronic or critically ill patients. In cancer patients, especially in patients with gastric cancer and pancreatic cancer, about 70% of patients will develop such symptoms. At the end of the cancer, about 80% of the patients will have cachexia symptoms, which is characterized by the fact that even if the patient increases the amount of food intake or increases the intake of nutrients, it will not prevent or prevent the patient's weight. keep falling. Hormone appetite promoters (such as Megestrol, medroxyprogesterone acetate) are often used in therapy, but in fact some people think that it is a pause in weight gain (only water and fat increase), body muscle mass does not increase, activity and physical strength are not seen. improve. Conversely, some side effects such as blood clots, edema, hemorrhage, high blood sugar, and high blood pressure can occur.

Catabolic consumption or cachexia is characterized by involuntary progressive consumption of fat and skeletal muscle, refractory weight loss to increase nutrient input, increase in static energy expenditure (REE), decreased protein synthesis, and changes in carbohydrate metabolism (Ke Increased activity of the Cori cycle), excessive catabolism of the muscle-dissolved ATP-ubiquitin-dependent proteasome pathway, and syndrome of adipose tissue catabolism through fat dissolution (Body JJ, Curr Opin Oncol) 11: 255-60, 1999, Muscaritoli M, et al: Eur J Cancer 42: 31-41, 2006). Typically, cachexia is diagnosed after the patient has reduced at least 5% or 5 pounds of pre-clinical weight. About half of all cancer patients experience some degree of catabolic consumption, and higher incidences are seen in cases of lung, pancreas, and gastrointestinal malignancies (Dewys WD, et al: Am J Med 69:491-7, 1980). . The syndrome can also be found in immunocompromised diseases such as AIDS patients and in patients suffering from bacterial and parasitic diseases, rheumatoid arthritis and chronic diseases of the intestines, liver, kidneys, lungs and heart. The cachexia is also associated with anorexia, which can be caused by aging or by physical injuries and burns. Cachexia syndrome reduces the patient's functional ability and quality of life, exacerbating the underlying condition and reducing tolerance to drugs. The degree of cachexia is inversely proportional to the patient's survival time, which usually represents a poor prognosis. In recent years, age-related diseases and disability have become major health concerns and importance.

Anorexia (a medical term for loss of appetite) is a manifestation of the deterioration of many malignant diseases and can be found in cancer, infectious diseases, chronic organ failure and traumatic patients. Anorexia is a serious syndrome because it leads to reduced calorie intake and malnutrition. Anorexia manifests itself in the loss of taste and smell of food, early satiety, reduced hunger and even total aversion to food, as well as symptoms of nausea and vomiting. Little is known about the causes of anorexia; effective treatment options are limited. Some studies indicate that a combination of hormonal, social, and psychological factors may be an important factor in the development and progression of the syndrome.

Although in fact, cachexia is often associated with cancer, it has not been confirmed that there is a consistent relationship between the occurrence of cachexia and the size or stage of tumor size, stage of disease, and malignant disease. However, cancer cachexia is often associated with decreased calorie intake, increased static energy expenditure, and altered protein, fat, and carbohydrate metabolism. For example, some significant abnormalities in carbohydrate metabolism include increased total glucose conversion rate, increased liver gluconeogenesis, glucose intolerance, and elevated blood glucose. Increased fat solubilization, increased conversion of free fatty acids and glycerol, hyperlipidemia, and decreased lipoprotein lipase activity are also frequently observed. Importantly, weight loss associated with cancer cachexia is not only due to reduced body fat storage, but also to the reduction in total body protein mass and extensive skeletal muscle consumption. Increased protein turnover and poor regulation of amino acid oxidation may also be important factors in the deterioration of this syndrome. In addition, in response to specific host-derived factors produced by cancer, for example, pre-inflammatory cytokines (tumor necrosis factor-α (TNF-α), interleukin-1, interleukin-6, and γ-interferon), Acute phase proteins (such as C-reactive protein), as well as specific prostaglandins, also appear to be associated with cancer cachexia.

In traditional Chinese medicine, it is recommended that the elderly should be given Chinese medicine every day. In addition to being prone to illness, the elderly can help to delay aging and live longer.

The applicant of the present application, in EP 1535619 A1, discloses a pharmaceutical composition for boosting human immunity. The composition contains a lanostane compound as an active ingredient, which enhances immunity and is used for prevention and treatment of viral infection. On the other hand, sputum extract and its purified (lanostanes) can inhibit immunity and are used for the prevention and treatment of allergic (asthmatic) caused by IgE. This inhibition is disclosed in the patent application No. 97123189 and PCT/CN2008/001218. . From these two patent applications it is shown that the extract of cockroach and its lanostanes can modulate immunity. Applicants of the present invention have disclosed that the human intestinal cells can enhance the absorption of nutrients by the action of the sputum extract and its purified lanostanes in the Chinese Patent No. 97111791 and the PCT/CN2008/000749 patent application No.; glucose, amino acid, Absorption of vitamins (folic acid).

Therefore, we believe that the extract components of sputum, especially lanostanes, may contribute to the improvement or treatment of cachexia related to nutrition and immunity. In the present invention, a human lung cancer cell is transplanted into a mouse to test whether the sputum extract component and the lanosterol compound have therapeutic effects on cachexia, and whether the human lung cancer cell transplanted mouse has the same normal appetite as the normal mouse. The weight is also normal and there is no gradual reduction.

The main object of the present invention is to provide a pharmaceutical composition for the treatment of the prevention and treatment of a wasting disease, such as cachexia and anorexia, especially a cachexia caused by cancer.

Another object of the present invention is to provide a sputum extract prepared from sputum for use in the treatment of the prevention and treatment of a wasting disease, such as cachexia and anorexia, especially a cachexia caused by cancer.

Another object of the present invention is to provide a new use of lanostane for the treatment of the prevention and treatment of wasting diseases such as cachexia and anorexia, especially cancer-caused cachexia.

The present invention discloses a pharmaceutical composition for preventing and treating a wasting disease (such as cachexia and anorexia) in a mammal (for example, a human), comprising a wool having the following chemical formula (I) as an active ingredient for preventing and treating an effective amount of a wasting disease. Sterol or its medicinal salt:

Wherein R ₁ is H or CH ₃ ; R ₂ is OCOCH ₃ , =O or OH; R ₃ is H or OH; and R ₄ is -C(=CH ₂ )-C(CH ₃ ) ₂ R _a , wherein R _a Is H or OH, or -CH=C(CH ₃ )-R _b , wherein R _b is CH ₃ or CH ₂ OH; R ₅ is H or OH; and R ₆ is CH ₃ or CH ₂ OH.

Preferably, the lanosterol (I) has the following chemical formula:

Preferably, the composition of the present invention contains from 0.1 to 60% by weight of lanosterol (I) or a pharmaceutically acceptable salt thereof.

Preferably, the compositions of the invention are administered orally.

A further object of the invention is to use a guanidine extract as a source of the lanosterol (I), the bismuth extract comprising from 1 to 60% by weight of the lanosterol of the above formula (I) and substantially free of Ring lanosterol.

Preferably, the hydrazine extract is prepared by a process comprising the following steps:

a) extracting the metabolite of the fungus, the fermentation product of the fungus or the hyphae of the fungus by using water, methanol, ethanol or a mixed solvent thereof;

b) concentration step a) extracting the obtained liquid;

c) introducing the concentrate obtained in step a) into a rubber hose column;

d) flushing the cartridge column with a low polarity eluting agent (eluent) and collecting the resulting eluate;

e) Concentration of the eluent of step d).

Preferably, the concentrate of the eluate obtained from step e) is analyzed by silica gel thin layer chromatography with a chromatographic value. The ultraviolet light and iodine were used as the detection side, and the developing solution was dichloromethane: methanol = 96:4.

Preferably, the solvent used in the extraction of step a) is 95% alcohol.

Preferably, step a) comprises extracting the metabolite of the fungus, boiling yeast or the hyphae of the fungus by boiling water; adding a base to the aqueous extract to a pH of 9-11; separating the alkaline aqueous solution Adding an acid to the basic aqueous solution to a pH of 4-7 to produce a precipitate; separating the precipitate; extracting the precipitate with alcohol and separating the extract liquid.

Preferably, the concentrate of step b) is further extracted by a two-phase solvent of 95% v/v aqueous methanol solution and n-hexane in a volume ratio of 1:1; the methanol layer is separated; and the methanol layer is concentrated to obtain The concentrate was used as the feed to the cartridge column of step c).

Preferably, the low polarity stripping agent of step d) is a mixed solvent of dichloromethane and methanol in a volume ratio of 96.5:3.5.

Preferably, the mash extract comprises from 5 to 35% by weight of lanosterol (I).

Preferably, the composition of the present invention further comprises a nutrient such as glucose, amino acid, vitamins or a combination thereof.

Preferably, the wasting disease of the present invention refers to cachexia; more preferably, the cachexia is caused by cancer, such as lung cancer, gastric cancer, pancreatic cancer, rectal cancer, breast cancer, oral cancer or nasopharyngeal cancer.

Preferably, the wasting disease of the present invention is caused by cancer, anorexia, aging, physical injury or burns.

Preferably, the pharmaceutical composition of the present invention is administered to a human in a dose of not less than 8.4 mg per day of the lanosterol (I).

Preferably, the pharmaceutical composition of the present invention further comprises a pharmaceutically acceptable carrier or diluent.

The present invention uses lanosterol of the formula (I) or a pharmaceutically acceptable salt thereof, or the aforementioned sputum extract as a treatment for loss of appetite and severe weight loss in cancer patients. In addition to cancer, diseases associated with cachexia include cachexia caused by AIDS, aging, rheumatoid arthritis, tuberculosis, fibrocysts, Crohn's disease, and infectious diseases. Surgical or cancer patients (with chemotherapy or radiation therapy) cause weakness and urgent need to supplement nutrients (amino acids, glucose, vitamins) to improve. The active ingredient of the present invention can be added to milk powder, beverage, food as a nutritional supplement purpose, or can be used as a medicine, such as a tablet, a capsule, a granule, a liquid, an injection, etc., for medical purposes.

Caclexia indicates that the human body is in a debilitating state, causing the cause of cachexia. There are many different opinions, but there is no exact mechanism. What can be known at present is that in cancer, cancer cell release factors or human immune response to cancer cells release various factors directly or indirectly cause anorexia, and the endocrine and immune system of the body changes. As a result, the body's fat and muscle protein are gradually reduced, resulting in patients who do not like diet, the body is getting weaker and the weight is gradually decreasing.

Diseases associated with cachexia include cancer, infectious diseases (such as tuberculosis, AIDS), autoimmune diseases (rheumatoid arthritis), the elderly, fibrocysts, and Crohn's disease. In cancer patients, especially those with stomach cancer and pancreatic cancer, about 70% of patients will have such symptoms. In addition, at the end of the cancer, about 80% of patients with any type of cancer will have cachexia symptoms. In the treatment, such as increased intake or other nutrients through feeding or intravenous injection, it is also impossible to prevent or prevent the patient's weight from decreasing. Hormonal appetite promoters (such as Megestrol, medoxy progesterone) are often used in therapy, but in fact some people think that it is a pause in weight gain (only water and fat increase), the body muscle mass does not increase, and the mobility is not improved. Conversely, some side effects such as blood clots, edema, hemorrhage, high blood sugar, and high blood pressure can occur.

A suitable method for preparing an active ingredient for treating cachexia from hydrazine disclosed in the present invention is, for example, the method disclosed in the aforementioned EP 1535619 A1, which comprises extracting hydrazine by a conventional extraction method to obtain a crude extract, and then separating into polar groups by chromatography. Small lanostane-like parts (with dichloromethane:methanol (96:4) as the extract) and polar keolanostane (with dichloromethane:methanol (90) :10 or 0:100) is the extract), wherein the position of the lanostane-like part is shown by the thin layer chromatography of tannin, that is, the developing solvent is dichloromethane-methanol (96:4). When the chromatographic value (Rf) is As for the open-chain smolanostane component, the chromatographic value is less than 0.1. The lanosterol moiety can be further separated by a gel column chromatography in which several lanostane compounds are isolated using dichloromethane:methanol (97:3 to 95:5).

The present invention will be further described in detail below with reference to the embodiments, but without the invention.

Embodiment 1

In Yunnan, 26 kg of pupa, 260 liters containing 75% alcohol, heated three times, combined with alcohol extract, concentrated under reduced pressure to obtain 225.2 grams of extract. The extract was quantitatively analyzed to obtain 76.27 mg of lanostanes per gram of extract, of which K1 (Pachymic acid) 33.4 g, K1-1 (dehydropachymic acid) 9.59 g, K2-1 (Tumulosic acid) ) 19.01 gram, K2-2 (dehydrotumulosic acid) 6.75 gram, K3 (Poylporenic acid C) 5.06 mg, K4 (3-epidehydrotumulosic acid) 2.46 gram.

Embodiment 2

Example 1 125 g of the alcohol extract was extracted 6 times with 1.3 liters of dichloromethane, and the dichloromethane extract was combined. After concentration, 22.26 g of the extract was obtained. The dichloromethane extract was dissolved in 95% hot alcohol, and the insoluble matter was filtered after being allowed to cool. The filtrate was added in a small amount of water until the alcohol content was 45%. At this time, a precipitate was formed, and the precipitate was obtained by centrifugation to obtain 17.4 g. Precipitate. Quantitative analysis showed that each gram of precipitate contained 264.78 mg of lanostanes, of which K1-1 (pachymic acid) 159.7 mg, K1-2 (dehydropachymic acid) 56.96 mg, K2-1 (Tumulosic acid) 24.43 mg , K2-2 (dehydrotumulosic acid) 8.8 mg, K3 (polyporenic acid) 9.84 mg, K4 (3-epidehydrotumulosic acid) 5.05 mg. The precipitate was tested by silica gel thin layer chromatography to prove that it did not contain ring-opened lanosterol.

Embodiment 3

After immersing 100 kg of medicinal herbs in 800 kg of water for 3 hours, the mixture was allowed to cool to 50 ° C, and the solution was adjusted to pH 11 with 5 N NaOH, and the solution was stirred for 3 hours. The liquid and solid were then separated by a centrifuge, and the solid was further added with 800 kg of water. The mixture was adjusted to pH 11 with NaOH as described above, stirred and centrifuged to remove the solid. The two liquids were combined, and the liquid was concentrated in vacuo to a 100 kg solution at 50 ° C, and then 3N HCl was added to pH 6.5 to give a precipitate. The precipitate was separated, washed with 40 L of H ₂ O, and then the precipitate was separated by a centrifuge, and sprayed with 8 L of water to obtain about 380 g of a powder. The powder was extracted three times with 4 L of alcohol, and the extracts were combined and concentrated to give 238.9 g of an alcohol extract. The extract was tested by silica gel thin layer chromatography (TLC) to prove that it did not contain ring-opened lanosterol. The extract was further separated by HPLC, and the extract was obtained as a main component of K2 214 mg, K3 23 mg, K4 24 mg and a small amount of K1 4.52 mg per gram of the extract, i.e., the extract contained about 265 mg of lanostanes per gram.

Or extract the powder with 4L of 50% alcohol aqueous solution, remove 50% alcohol aqueous solution and collect insoluble powder, and repeat three times to obtain 245.7g of 50% alcohol aqueous solution insoluble matter. The TLC method indicates that the insoluble matter does not contain open loop. The lanosterol is further purified by HPLC to separate the insoluble matter. The main component is K2 214 mg, K3 23 mg, K4 24 mg and a small amount of K1 4.52 mg per gram, that is, the extract contains about 261 mg of lanostanes per gram.

Embodiment 4

After 30 kg of glutinous rice in Yunnan, it was ground into powder and extracted with 120 L of alcohol (concentration 95%) for 24 hours, and separated by filtration. The foregoing extraction and solid-liquid separation were repeated three times. The filtrate was combined and concentrated to give a dry extract 265.2 g. The dried extract was subjected to partition extraction using a two-phase extractant (hexane: 95% methanol = 1:1). The methanol layer was taken out and concentrated to give 246.9 g of dry solid. The dried solid was separated by a silica gel column chromatography, which was filled with 10-40 times the weight of the dry solid, which was purchased from Merck, Silica gel 60, 70-230 mesh. The mixture of dichloromethane/methanol was used as a solvent (eluent), and the mixture was extracted in a ratio of 96:4, 90:10, and 0:100, and the eluate was subjected to silica gel thin layer chromatography. (Thin Layer Chromatography) (UV light and iodine detection, the developing solution is dichloromethane: methanol = 96: 4) The components are detected, and the same components are combined.

Chromatography column chromatography using a dichloromethane-methanol (96:4) mixture gave 78 g of the PCM portion of the hydrazine extract of the present invention. The PCM part can clearly see 6 traces according to the above-mentioned silica gel thin layer chromatography. The combined chromatography of dichloromethane:methanol (90:10) and (0:100) gave 168 g of PCW fraction.

The PCM fraction is further chromatographed with dichloromethane:methanol (96.5:3.5) as a stripping agent (same as the above-mentioned ruthenium column), and further purified lanostane-like component K1 (K1) -1 and K1-2), K2 (K2-1 and K2-2), K3, K4, K4a, K4b, K5, K6a and K6b. Detailed separation steps and identification analysis data can be found in EP 1535619 A1.

The above K1 to K6b compounds have the following structure:

The yields of the lanosterol compounds K1 to K6b separated from the PCM fraction are shown in the following table. The PCM fraction contains about 15% by weight of lanosterol compounds K1 to K6b.

Example 5: Preparation of capsules

A capsule containing the hydrazine extract PCM component prepared in Example 4 was prepared according to the following composition:

The cockroach extract PCM and sodium strontium aluminate were respectively sieved through a #80 mesh sieve, the potato starch was sieved through a #60 mesh sieve, and magnesium stearate was sieved through a #40 mesh sieve, and then placed. The mixer was stirred well and then filled into vacant capsules, each containing about 1.68 mg (0.42% by weight) of active ingredient K1-K6.

Embodiment 6

The cockroach extracts prepared in Example 2 and Example 3 of the present invention were respectively formulated with the medicinal agents shown in Table 1 as test substances to evaluate changes in body weight, food intake, and amount of protein in the serum caused by cancer, and cancer. The overall nutritional status of the sputum extract extract taken by the cachectic patient.

Example 7

This example is an animal test for treating cachexia caused by cancer with the agent of Table 1. Human lung cancer cells were transplanted into mice to demonstrate whether the extract of sputum can have a therapeutic effect on cachexia. Observe the changes of body weight, food intake and serum albumin concentration in human lung cancer cell-transplanted mice and normal mice to evaluate its treatment. The efficacy of cancer cachexia.

Experimental animal

The test mice were purchased from the National Taiwan University Animal Center for six to eight weeks of age. CB-17 SCID mice were housed in stainless steel cages. Room temperature is controlled at 25 ± 2 ° C, humidity range is 40-70%, 12 hours light / dark alternate, drinking water is not limited. After the purchase, the adaptation period was first given in the animal room. After weighing the machine, the similar weights were grouped into the same group in random numbers. Statistical analysis confirmed that there was no significant difference in body weight between the groups.

Culture of human lung cancer cell line H460

The preserved H460 cell line was taken out from the liquid nitrogen tube, thawed in a 37 ° C water bath, and 8 ml of 10% FBS DMEM medium was added to the console, and centrifuged at 1200 rpm for 10 minutes. After removing the supernatant, the cells were added. 8 ml of the culture solution was cultured in an incubator at 37 ° C, 5% CO ₂ until the desired amount of cells.

Mouse orthotopic implantation of lung cancer cell line H460

After deep anesthesia with pentobarbital, a 0.5 ml insulin syringe was used to implant 0.1 ml (1×10 ⁶ /ml) of H460 cell line into the thoracic cavity of the mouse. After the cells are implanted, the mice are returned to the original mouse cage and allowed to recover by themselves.

Drug treatment

The cockroach extract prepared in Example 2 was weighed 67.7 mg to be added to the sterile water to 10 ml, and the extract was suspended in a solution to form a PC-A agent, and 5 ml of the PC-A agent was sterilized by ultrasonic treatment. The amount of water was quantified to 10 ml, which was a PC-B agent, and 5 ml of PC-B agent was quantified in sterile water to 10 ml to form a PC-C agent. The mice were fed according to their respective body weights, and the 0.25 cc dose of the drug was administered to each mouse in a 25 gm tube. Dosage. The test was performed in two divided sessions; the normal mouse group (blank) was fed normal mice with distilled water (substituted with culture medium PBS instead of H460 injected into the thoracic cavity). In the control group, lung cancer mice were fed with distilled water. The feeding method is to connect the 18th (5 cm long) steel pipe with a syringe (1 cc), hold the mouse double mouth with the left hand while feeding, and then gently pipe the tube into the stomach, then feed the medicine or distilled water. , each volume is limited to not more than 0.3cc.

The first trial was divided into normal group (blank), control group (control group) and three groups of administration groups (different lanosterol doses), a total of five groups of 9 mice per group. Three groups of the drug-administered group were given PC-A, PC-B and PC-C of the cockroach extract prepared in Example 2, and the lung cancer mice were administered with sterol per kilogram body weight of 17.6 mg, 8.8. Mg and 4.4 mg, as shown in Table 1.

The second test was divided into a normal group (blank), a control group and a drug-administered group, and a total of three groups of three mice each. The drug-administered group was administered with lung cancer mice to the drug PC-D fed the sputum extract prepared in Example 3. The mice were administered 8.8 mg of lanosterol per kg of body weight, as shown in Table 1.

Serum albumin blood sample collection

Whole blood was collected from the corner of the mouse on the 22nd day, and the blood sample was allowed to stand at room temperature for 1 hour. The cells were centrifuged twice at 3,000 rpm, serum was collected, and stored at -20 ° C, and the serum albumin content was analyzed in the future.

Determination of serum albumin concentration

Analytical reagent kit analysis was performed using a Mouse albumin ELISA kit (Bethyl Laboratories Inc., Texas, US). Anti-mouse albumin was diluted 100-fold with TBS, and 100 μl/well well was coated in a microtiter plate and allowed to act at room temperature for 1 hour. After washing 3 times with TBST, blocking was performed with a 1% BSA 200 μl/well well, allowed to stand at room temperature for 1 hour, and then washed 3 times with TBST. Add the sample and standard solution, apply for 1 hour at room temperature, wash 5 times with TBST, and add 100 μl/well well of goat anti-mouse albumin-HRP conjugate. 1% BSA and 0.05% Tween 20 TBS were diluted 10,000 times) for 1 hour at room temperature. Wash 5 times with TBST, add 100 μl of substrate solution TMB (3,3',5,5'-tetramethylbenzidine), avoid the reaction for 15 minutes at room temperature, and add 100 μl/well well of 2N HCl to stop the reaction and determine A _450nm. The absorbance value.

statistical methods

Data results are expressed as mean ± standard deviation. The statistical principle firstly uses one-way ANOVA to check the variation between groups. If P<0.05, Dunnett's multiple range t-test is used to compare the differences between groups. The control group separately compares the principles.

Weight and food intake changes

The changes in food intake and body weight changes of each group of mice in the first and second trials were observed.

result

Mouse weight change

In the first experiment, the change in body weight during lung tube feeding in lung cancer mice is shown in Figure 1. The control mice showed a significant decrease in body weight after implantation of H460 lung cancer cells. Compared with mice implanted with lung cancer cells, low doses of PC-C (4.4 mg/kg) and PC-B were given to the sputum extracts. 8.8mg/kg), there is no difference between the two, that is, no improvement in cachexia symptoms. However, if the dose is increased to 17.6 mg/kg (PC-A), the decrease in body weight of the mice is obviously slowed down. Therefore, the sputum component can improve the weight loss caused by the implantation of H460 lung cancer. A second experiment was performed using different sputum extracts, extract PC-D (8.8 mg/kg), and the body weight changes in lung cancer mice are shown in Figure 2. As shown in Figure 1, the control mice showed a significant decrease in body weight after implantation of H460 cells, whereas normal mice (Blank) (not implanted with H460 cells) gained slightly more weight, while PC-D was given sputum extract. The group found that it was able to maintain the body weight at the start of the trial, so it was possible to prove that it can treat cachexia.

Feed intake changes in cancer mice

The change in food intake of lung cancer mice after tube feeding of sputum extract is shown in Figure 3. In the control mice, food intake gradually decreased after implantation of H460 lung cancer cells. Compared with mice administered with lung cancer cells and then given sputum extract (PC-D), the intake of food did not show a gradual decline, but it was the same as that of normal mice (not implanted with H460). There was no difference in the amount of food intake between the two groups of mice. Therefore, sputum extract can improve or treat the disease caused by cachexia caused by poor feeding.

Changes of albumin content in serum of lung cancer mice

The changes in serum albumin content of each group of mice in the first and second trials are shown in Figure 4 and Figure 5. The mice in which the control group was implanted with lung cancer cells H460) had a tendency to decrease serum albumin relative to the normal group (Blank) mice (not inoculated with H460). In the administration group (PC-A, B, C) fed with cockroach extract, it was observed that the increase in the dose of lanolin increased the albumin content in the blood of the mice, and the dose of 17.6 mg/kg was given to PC-A. A significant increase was observed. The same significant effect was also obtained in the administration group PC-D (8.8 mg/kg dose) to which the sputum extract was administered.

Since the animal body produces an inflammatory response to lung cancer cells, this reaction changes the production of hepatocyte proteins, some proteins reduce manufacturing, and some proteins (or induced proteins) are produced. Therefore, in addition to observing changes in body weight and food intake, another method can measure albumin in the blood, and albumin in the blood decreases, indicating that the animal is affected by lung cancer cells, so that the albumin produced by the liver cells is reduced, so it is released into the blood. The albumin is reduced. As shown in Fig. 4 and Fig. 5, the blood albumin content of cachexia mice decreased, but the blood albumin content of the cachexia mice administered with sputum extract did not decrease, and the sputum extract obviously had the effect of treating cachexia. .

Example 8:

Human clinical studies on the treatment of cancer-caused cachexia by sputum extract:

(1) Observing the improvement of the overall nutritional status of the patient.

(B) to observe the improvement in patient weight.

(A) Grouping and administration of cancer patients:

15 cancer patients sustained weight loss, belonging to gastric cancer (3), pancreatic cancer (3), rectal cancer (3), breast cancer (4), oral cancer (1), and nasopharyngeal carcinoma (1) ), randomly divided into 3 groups of 5 people each, the first group was given a low dose of sputum extract (extracted from Example 3, 16.8 mg / each), one capsule per day. The second group was given a higher dose of sputum extract, two capsules per day. The third group did not give sputum extracts only to take chemotherapy drugs for comparison purposes.

(B) Treatment: 15 patients received chemotherapy for 6 weeks, the first/second group took sputum and took 4 weeks (5/6 weeks only chemotherapy), and the ‧ body weight/nutrition status assessment form was recorded weekly.

(C) Treatment outcome (1): The weight of the cancer patient is improved. After the treatment (sixth week or 42 days later), the patient's body weight is compared with the treatment day 1 body weight as follows:

From the treatment results, it can be seen that the chemotherapeutic drug and the sputum extract group (one/two group) are better than the third group of the control (only chemotherapy is given without sputum), and the body weight can be Weight loss was observed to be still below the weight of day 1 of treatment.

(D) Treatment results (2) Improvement of overall nutritional status of patients

The overall nutritional status of cancer patients was assessed based on the Patient-Generated Subjective Global Assessment used by clinicians to assess (1) weight improvement, (2) dietary improvement, and (3) improvement in symptoms and symptoms. (4) The comprehensive score of physical fitness improvement, as shown in Figure 6, Figure VII and Figure 8 below, Figure 6 is the first group of patients (1 capsule of chemotherapy + sputum extract capsule), and Figure 7 is the second group of patients (chemotherapy + 2 capsules of sputum extract, Figure 8 is the control group (chemotherapy). The lower the PG-SGA score, the better the patient's improvement procedure and the overall health trend.

From the treatment results, the improvement of the health/life quality of patients with cachexia treated with sputum extract and chemotherapy drugs far exceeds the chemotherapy-only cachexia patients, and it is statistically significant.

From the results of this example, it can be found that the combination of the sputum extract and the chemotherapeutic drug can prevent the weight loss of the cancer patient, and the overall nutritional status of the patient is improved, which is significantly better than the cancer patient who only receives chemotherapy.

Figure 1 shows the change in body weight of each group of mice in the first trial of Example 7, where + represents the normal group (blank); black dots represent the control group; diamonds, squares, and triangles represent the drug-administered group, respectively. PC-A, PC-B and PC-C, mice were administered with sterol 17.6 mg, 8.8 mg and 4.4 mg per kilogram of body weight, respectively.

Figure 2 shows the change in body weight of each group of mice in the second test of Example 7, wherein the diamond represents the normal group; the square represents the control group; and the triangle represents the drug group PC-D, the mouse Rhenol 8.8 mg was administered per kg of body weight.

Figure 3 shows the change in food intake of each group of mice in the second experiment of Example 7, wherein the diamond represents the normal group; the triangle represents the control group; and the circle represents the drug group PC-D, Mice were administered 8.8 mg of sterol per kg of body weight.

Figure 4 shows the albumin concentration in the blood of day 22 of each group of mice in the first trial of Example 7.

Figure 5 shows the albumin concentration in the blood of day 22 of mice of each group of mice in the second experiment of Example 7.

Figure 6 is an evaluation of the overall nutritional status of the first group of cancer patients (1 chemotherapy + sputum extract capsule) of Example 8 based on the Patient-Generated Subjective Global Assessment. to evaluate.

Figure 7 is an estimate of the overall nutritional status of a second group of cancer patients (2 chemotherapy + sputum extract capsules) of Example 8 based on the PG-SGA seduce used by clinicians.

Figure 8 is an evaluation of the overall nutritional status of a control group of cancer patients (chemotherapy) in Example 8, which is based on the PG-SGA scale used by the clinician.

Claims (11)

Use of a lanosterol having the following chemical formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating cachexia caused by cancer in a mammal, Wherein R ₁ is H or CH ₃ ; R ₂ is OCOCH ₃ , =O or OH; R ₃ is H or OH; and R ₄ is -C(=CH ₂ )-C(CH ₃ ) ₂ R _a , wherein R _a is H or OH, or -CH=C(CH ₃ )-R _b , wherein R _b is CH ₃ or CH ₂ OH; R ₅ is H or OH; and R ₆ is CH ₃ or CH ₂ OH. The use of the first aspect of the patent application, wherein the lanosterol (I) has the following chemical formula: The use of claim 1, wherein the medicament contains from 0.1 to 60% by weight of lanosterol. The use of the first aspect of the patent application, wherein the medicament is orally administered. The use of the first aspect of the patent application, wherein the mammal is a human. The use of claim 1, wherein the medicament comprises a monoterpene extract comprising from 1 to 60% by weight of lanosterol of formula (I) and substantially free of ring-opened lanosterol. The use of claim 6 wherein the bismuth extract comprises from 5 to 35% by weight of lanosterol (I). The use of the scope of claim 6 wherein the lanosterol (I) has the following chemical formula: The use of the first aspect of the patent application, wherein the cancer is lung cancer, gastric cancer, pancreatic cancer, rectal cancer, breast cancer, oral cancer or nasopharyngeal cancer. The use of the fifth aspect of the patent application, wherein the daily dose of the drug is lanosterol (I) and the daily dose is not less than 8.4 mg. The use of claim 1, wherein the medicament further comprises a pharmaceutical acceptable carrier or diluent.