A composition of lotus leaf alkaloids for preventing and treating obesity of pet dogs, and a preparation method and application thereof

Abstract

The patent discloses a kind of lotus leaf base compositions for preventing and treating pet dog obesity, which is made of the following raw medicinal materials by weight: lotus leaf base 5-10 parts, egg yolk lecithin 50-100 parts, beta-sitosterol 10-20 parts, and sodium alginate 10-20 parts. The lotus leaf base is first encapsulated in liposomes with egg yolk lecithin and beta-sitosterol as the lipid film, and then the prepared lotus leaf base liposomes are wrapped with sodium alginate. Compared with the traditional method, the lotus leaf base composition provided by the present application not only provides double-layer protection and sustained-release effect for lotus leaf base absorption, but also provides targeted delivery effect for lotus leaf base. In addition, it can also reduce the absorption of cholesterol by pet dogs. The lotus leaf base composition prepared by the present application has good compatibility with pet dog tissues, can maintain bioavailability for a long time, and has excellent preventive and therapeutic effects on pet dog obesity.

Description

Technical Field

[0001] This invention relates to a pet product, and more specifically, to a lotus leaf alkaloid composition for preventing and treating obesity in dogs and its preparation method. Background Technology

[0002] With the increasing popularity of pet dogs in my country, the incidence of canine obesity is also rising year by year. Currently, treatment options for canine obesity remain very limited, relying heavily on methods used in human obesity prevention and treatment (such as weight loss medication, surgical weight loss, intermittent fasting, low-calorie diets, and exercise), with less than ideal results. Therefore, weight loss for pet dogs is a very challenging task, and the development of safe, green, and convenient weight loss products has broad market prospects.

[0003] Lotus leaf alkaloid is an active ingredient extracted from the dried leaves of the lotus plant. It has effects such as weight loss, lipid reduction, anti-oxidation, and blood sugar reduction, and has been widely used in food, health products, and pharmaceuticals in recent years. However, its relatively low solubility and poor bioavailability have hindered the development of lotus leaf alkaloid as an anti-obesity drug or health product for dogs.

[0004] Chinese patent CN111149937A discloses a fresh meat nutritional paste for controlling the weight of pet cats. This paste contains lotus leaf extract, which can effectively control obesity in cats and restore them to a healthy body shape. However, the patent does not identify the components of the lotus leaf extract, and therefore cannot fully demonstrate that lotus leaf alkaloids are responsible for its effect, as flavonoids in lotus leaf extract have weight-loss and lipid-lowering effects.

[0005] Chinese patent CN115968977A discloses a weight-loss pet food and its preparation method. This pet food directly incorporates lotus leaf alkaloids, which have a certain weight-loss effect. However, studies have shown that lotus leaf alkaloids have poor solubility, low oral absorption, and are rapidly absorbed and eliminated in canines with a short residence time, which reduces their bioavailability. Therefore, directly adding lotus leaf alkaloids to pet food may affect its efficacy.

[0006] Chinese patent CN115487205A discloses a black tea polysaccharide-lotus leaf alkaloid microgel complex, its preparation method, and its application. In this complex, lotus leaf alkaloid liposomes are first prepared using lotus leaf alkaloid, soybean lecithin, and cholesterol, and then compounded with black tea polysaccharide / proanthocyanidin microgels. This provides dual protection and sustained-release for lotus leaf alkaloid, improving its bioavailability and efficacy. However, because this patent contains cholesterol, long-term use may lead to excessive intake, causing hypercholesterolemia, which can result in atherosclerosis, hypertension, and cardiovascular diseases, thus having certain limitations. Summary of the Invention

[0007] Therefore, it is necessary to provide a lotus leaf alkaloid composition for preventing and treating obesity in pet dogs and its preparation method, in order to address the above-mentioned technical problems.

[0008] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0009] A lotus leaf alkaloid composition for preventing and treating obesity in pet dogs, characterized in that the lotus leaf alkaloid composition for preventing and treating obesity in pet dogs is made from the following raw materials in parts by weight: 5-10 parts lotus leaf alkaloid, 50-100 parts egg yolk lecithin, 10-20 parts β-sitosterol, and 10-20 parts sodium alginate.

[0010] In a preferred embodiment of the present invention, the weight ratio of egg yolk lecithin to β-sitosterol is 5:1 to 8:1.

[0011] In a preferred embodiment of the present invention, the weight ratio of egg yolk lecithin to β-sitosterol is 5.7:1.

[0012] In a preferred embodiment of the present invention, the weight ratio of the sum of egg yolk lecithin and β-sitosterol to lotus leaf alkaloid is 8:1 to 14:1.

[0013] In a preferred embodiment of the present invention, the weight ratio of the sum of egg yolk lecithin and β-sitosterol to lotus leaf alkaloid is 12:1.

[0014] A method for preparing a lotus leaf alkaloid composition for preventing and treating obesity in pet dogs, comprising:

[0015] Step S1: Take 5-10 parts of lotus leaf alkaloid, 50-100 parts of egg yolk lecithin and 10-20 parts of β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the concentration of egg yolk lecithin 5-10 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved.

[0016] Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:1 to 1:3. Then place it on a magnetic stirrer to hydrate for 30 min.

[0017] Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark.

[0018] Step S4: Take 10-20 parts of sodium alginate according to the mass fraction, dissolve them in PBS solution with pH 6.8-7.4 to prepare a 0.5 mg / ml sodium alginate solution;

[0019] Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition.

[0020] Step S6: Seal the lotus leaf alkaloid composition obtained in step S5 and store it at 4°C in the dark.

[0021] The present invention also discloses the application of the aforementioned lotus leaf alkaloid composition for preventing and treating obesity in pet dogs in the preparation of feed or feed additives that have the effect of preventing and treating obesity in pet dogs.

[0022] The present invention also discloses the application of the lotus leaf alkaloid composition for preventing and treating obesity in pet dogs prepared by the aforementioned preparation method in the preparation of feed or feed additives that have the effect of preventing and treating obesity in pet dogs.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] This invention provides a lotus leaf alkaloid composition for preventing and treating obesity in dogs. The invention combines lotus leaf alkaloid, which has low water solubility, with other substances, and utilizes liposome encapsulation and sodium alginate coating to significantly enhance the sustained-release effect and stability of the lotus leaf alkaloid, thereby improving its bioavailability. The lotus leaf alkaloid composition prepared by this invention exhibits good tissue compatibility with dogs, overcoming the disadvantage of rapid absorption and metabolism of lotus leaf alkaloid in canines, and can maintain bioavailability for a long time, demonstrating excellent preventive and therapeutic effects on canine obesity. Attached Figure Description

[0025] To more clearly illustrate the solutions in this invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0026] Figure 1 A bar graph showing the effect of lotus leaf alkaloid composition on the body weight of dogs on a high-fat diet;

[0027] Figure 2 A bar graph showing the effect of lotus leaf alkaloid composition on subcutaneous fat thickness in dogs on a high-fat diet;

[0028] Figure 3 This is a bar graph comparing the body weight of obese dogs before and after treatment with the lotus leaf alkaloid composition. Implementation

[0029] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, thereby providing a clearer and more explicit definition of the scope of protection of the present invention.

[0030] The lotus leaf alkaloid composition for preventing and treating obesity in pet dogs is made from the following raw materials in parts by weight: 5-10 parts lotus leaf alkaloid, 50-100 parts egg yolk lecithin, 10-20 parts β-sitosterol, and 10-20 parts sodium alginate.

[0031] Furthermore, the weight ratio of egg yolk lecithin to β-sitosterol is 5:1 to 8:1. Preferably, the weight ratio of egg yolk lecithin to β-sitosterol is 5.7:1.

[0032] Furthermore, the weight ratio of the sum of egg yolk lecithin and β-sitosterol to lotus leaf alkaloid is 8:1 to 14:1. Preferably, the weight ratio of the sum of egg yolk lecithin and β-sitosterol to lotus leaf alkaloid is 12:1.

[0033] The following describes the preparation method of this lotus leaf alkaloid composition, including:

[0034] Step S1: Take 5-10 parts of lotus leaf alkaloid, 50-100 parts of egg yolk lecithin and 10-20 parts of β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the concentration of egg yolk lecithin 5-10 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved.

[0035] Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:1 to 1:3. Then place it on a magnetic stirrer to hydrate for 30 min.

[0036] Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark.

[0037] Step S4: Take 10-20 parts of sodium alginate according to the mass fraction, dissolve them in PBS solution with pH 6.8-7.4 to prepare a 0.5 mg / ml sodium alginate solution;

[0038] Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition.

[0039] Step S6: Seal the lotus leaf alkaloid composition obtained in step S5 and store it at 4°C in the dark.

[0040] The following are further examples illustrating this preparation method. Example

[0041] Step S1: Take 7 parts by weight of lotus leaf alkaloid, 72 parts by weight of egg yolk lecithin and 13 parts by weight of β-sitosterol, add them to anhydrous ethanol solution to make the concentration of egg yolk lecithin 10 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved.

[0042] Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:3. Then place it on a magnetic stirrer to hydrate for 30 min.

[0043] Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark.

[0044] Step S4: Take 15 portions of sodium alginate according to the mass fraction, dissolve them in PBS solution with pH 7.4 to prepare a 0.5 mg / ml sodium alginate solution;

[0045] Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition.

[0046] Step S6: After sealing the lotus leaf alkaloid composition obtained in step S5, store it at 4°C in the dark. Example

[0047] Step S1: Take 5 parts lotus leaf alkaloid, 55 parts egg yolk lecithin and 15 parts β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the egg yolk lecithin concentration 8 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved.

[0048] Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:3. Then place it on a magnetic stirrer to hydrate for 30 min.

[0049] Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark.

[0050] Step S4: Take 10 portions of sodium alginate according to the mass fraction, dissolve them in PBS solution with pH 7.4 to prepare a 0.5 mg / ml sodium alginate solution;

[0051] Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition.

[0052] Step S6: After sealing the lotus leaf alkaloid composition obtained in step S5, store it at 4°C in the dark. Example

[0053] Step S1: Take 10 parts lotus leaf alkaloid, 90 parts egg yolk lecithin and 20 parts β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the egg yolk lecithin concentration 5 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved.

[0054] Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:3. Then place it on a magnetic stirrer to hydrate for 30 min.

[0055] Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark.

[0056] Step S4: Take 15 portions of sodium alginate according to the mass fraction, dissolve them in PBS solution with pH 7.0 to prepare a 0.5 mg / ml sodium alginate solution;

[0057] Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition.

[0058] Step S6: After sealing the lotus leaf alkaloid composition obtained in step S5, store it at 4°C in the dark. Example

[0059] Step S1: Take 8 parts of lotus leaf alkaloid, 100 parts of egg yolk lecithin and 16 parts of β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the concentration of egg yolk lecithin 6 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved.

[0060] Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:3. Then place it on a magnetic stirrer to hydrate for 30 min.

[0061] Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark.

[0062] Step S4: Take 10 portions of sodium alginate according to the mass fraction, dissolve them in PBS solution with pH 6.8 to prepare a 0.5 mg / ml sodium alginate solution;

[0063] Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition.

[0064] Step S6: After sealing the lotus leaf alkaloid composition obtained in step S5, store it at 4°C in the dark. Example

[0065] Step S1: Take 8 parts of lotus leaf alkaloid, 85 parts of egg yolk lecithin and 16 parts of β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the concentration of egg yolk lecithin 8 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved.

[0066] Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:3. Then place it on a magnetic stirrer to hydrate for 30 min.

[0067] Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark.

[0068] Step S4: Take 15 portions of sodium alginate according to the mass fraction, dissolve them in PBS solution with pH 7.4 to prepare a 0.5 mg / ml sodium alginate solution;

[0069] Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition.

[0070] Step S6: After sealing the lotus leaf alkaloid composition obtained in step S5, store it at 4°C in the dark. Example

[0071] Step S1: Take 7 parts lotus leaf alkaloid, 80 parts egg yolk lecithin and 18 parts β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the egg yolk lecithin concentration 10 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved.

[0072] Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:3. Then place it on a magnetic stirrer to hydrate for 30 min.

[0073] Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark.

[0074] Step S4: Take 20 portions of sodium alginate according to the mass fraction, dissolve them in PBS solution with pH 7.4 to prepare a 0.5 mg / ml sodium alginate solution;

[0075] Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition.

[0076] Step S6: After sealing the lotus leaf alkaloid composition obtained in step S5, store it at 4°C in the dark.

[0077] The following comparative examples illustrate this preparation method.

[0078] Comparative Example 1

[0079] In this comparative example, the mass fractions of lotus leaf alkaloid, egg yolk lecithin, and β-sitosterol are the same as in Example 1. The only difference is that sodium alginate coating was not used in the preparation method.

[0080] Comparative Example 2

[0081] In this comparative example, the mass fractions of lotus leaf alkaloid and egg yolk lecithin are the same as in Example 1. The only difference is that β-sitosterol in Example 1 is replaced with the same mass fraction of cholesterol.

[0082] The following is a physicochemical analysis of the lotus leaf alkaloid compositions obtained in Examples 1 to 6.

[0083] Packet rate assessment:

[0084] Accurately weigh the lotus leaf alkaloid standard (i.e., the lotus leaf alkaloid compositions obtained in Examples 1 to 6), dissolve it in methanol to prepare a 100 μg / ml lotus leaf alkaloid standard solution. Take 0.5 ml of the lotus leaf alkaloid standard solution and dilute it to 5 ml with methanol. Using a blank solvent as a reference, perform a full wavelength scan at 200–500 nm to determine the maximum absorption wavelength of lotus leaf alkaloid. Accurately pipette 0.1, 0.3, 0.5, 1.0, 1.5, and 2.0 ml of the lotus leaf alkaloid standard solution, respectively, and dilute to 5 ml with methanol solution. Shake well and measure the absorbance. Perform linear regression with concentration (μg / ml) as the x-axis and absorbance A as the y-axis to obtain a standard curve. Take 8, 16, and 32 μg / ml of the lotus leaf alkaloid standard solution to investigate its recovery rate. Take 0.1 ml of lotus leaf alkaloid liposome suspension, dilute with methanol to 5 ml, sonicate to break the emulsion, and measure the absorbance at 270 nm. Calculate the lotus leaf alkaloid content according to the standard curve. Repeat the above experiment three times and take the average value. Calculate the encapsulation efficiency (EE) of lotus leaf alkaloid liposomes using the following formula: EE(%) = (mass of drug in liposomes / mass of drug added) × 100%.

[0085] Particle size and zeta potential determination:

[0086] The lotus leaf alkaloid liposomes in each example and comparative example were diluted 10-fold, and the particle size and zeta potential of the liposomes were measured using a particle size potentiometer. The results are shown in Table 1 below.

[0087] Table 1. Encapsulation efficiency, particle size, and zeta potential of lotus leaf alkaloid liposomes

[0088] Group Packet rate (%) Particle size (nm) zeta potential (mV) Example 1 83.14 103.43 -15.25 Example 2 82.31 105.18 -16.88 Example 3 79.46 106.36 -13.41 Example 4 80.33 98.14 -15.63 Example 5 78.95 110.22 -14.76 Example 6 79.08 97.59 -14.27 Comparative Example 1 68.42 94.01 5.16 Comparative Example 2 79.77 102.65 -15.83

[0089] As can be seen, the encapsulation efficiency of the lotus leaf alkaloid liposomes in all embodiments is greater than 78%, the particle size is around 100 nm, and the zeta potential is around -15 mV. This indicates that the preparation method used in this invention can obtain lotus leaf alkaloid liposomes with good performance, with Example 1 showing the best performance. The lotus leaf alkaloid liposomes in Comparative Example 1 have the lowest encapsulation efficiency and a positive zeta potential. This is because the lotus leaf alkaloid is embedded or adsorbed on the liposome surface and no secondary encapsulation with sodium alginate is used.

[0090] The following are relevant instructions regarding animal experiments.

[0091] Sixty-six adult Beagles, weighing (10±0.5) kg, were acclimatized to a normal diet for two weeks and then randomly divided into 11 groups: a normal diet group, a high-fat diet group, a lotus leaf alkaloid raw material group, and the remaining groups were designated as Examples 1-6 and Comparative Examples 1-2, respectively. The normal diet group was fed a normal diet, while the remaining groups were fed a high-fat diet. Dogs in Examples 1-6 and Comparative Examples 1-2 received 10 ml of their respective lotus leaf alkaloid liposomes orally daily, while dogs in the high-fat diet + lotus leaf alkaloid group received an equal amount of lotus leaf alkaloid raw material orally via gavage daily. The experiment lasted for 8 weeks, and daily food intake was calculated. After the experiment, body weight, abdominal subcutaneous fat thickness, and blood lipid levels were measured and statistically analyzed. The results are as follows: Figure 1 , Figure 2 As shown in Table 2 below,

[0092] Table 2. Effects of lotus leaf alkaloid liposomes on serum triglycerides and total cholesterol in dogs on a high-fat diet (n=6)

[0093] Group Serum triglycerides (mmol / L) Serum total cholesterol (mmol / L) Normal diet group 0.93±0.14 3.88±0.33 High-fat diet group 1.77±0.29 5.46±0.39 Lotus leaf alkali raw material group 1.52±0.26 5.02±0.32 Example 1 1.14±0.14 4.29±0.26 Example 2 Group 1.23±0.17 4.53±0.31 Example 3 Group 1.33±0.13 4.65±0.35 Example 4 1.25±0.14 4.38±0.32 Example 5 group 1.41±0.22 4.67±0.41 Example 6 group 1.31±0.19 4.62±0.40 Comparative Example 1 1.46±0.23 4.86±0.43 Comparative Example 2 1.43±0.20 5.32±0.37

[0094] It can be seen that there was no difference in the average food intake among all groups of dogs. All example groups reduced the weight, subcutaneous fat thickness, and blood lipid levels of dogs on a high-fat diet, with better results than the lotus leaf alkaloid raw material group. Example 1 showed the best effect, indicating that the weight loss effect of each example was not related to food intake but to the amount of subcutaneous fat accumulation. The weight and subcutaneous fat thickness of dogs in Comparative Example 1 were lower than those in the lotus leaf alkaloid raw material group but slightly higher than those in the example groups. The weight and subcutaneous fat thickness of dogs in Comparative Example 2 were comparable to those in the example groups. However, the serum total cholesterol level of dogs in Comparative Example 2 was higher than that in the example groups. This proves that the weight loss and lipid-lowering effect of the lotus leaf alkaloid liposomes of the present invention is superior to that of liposomes using lotus leaf alkaloids alone, liposomes not coated with sodium alginate, and liposomes using cholesterol as a lipid membrane. Furthermore, the lotus leaf alkaloid composition prepared in Example 1 showed the best effect in preventing obesity in pet dogs.

[0095] In addition, the lotus leaf alkaloid composition prepared in Example 1 above was used to intervene in the treatment of obese dogs, and its weight loss effect was evaluated. The method was as follows: From August to October 2022, five obese dog cases were collected from Taimu Animal Hospital (Taizhou City, Jiangsu Province). With the owner's consent, and under unchanged dietary conditions, lotus leaf alkaloid liposomes were administered orally at a dose of 1 ml / kg body weight, once daily, for eight consecutive weeks. Weight was then measured, and weight changes were observed.

[0096] The results are as follows Figure 3 As shown, after 8 weeks of treatment, all 5 dogs experienced varying degrees of weight loss, with a reduction of more than 10% compared to before treatment.

[0097] This lotus leaf alkaloid composition has many advantages:

[0098] 1) The lotus leaf alkaloid in this composition is a compound found in the lotus leaf, a natural and green plant with good weight loss and lipid-lowering effects. However, it is rapidly absorbed and metabolized in dogs, resulting in low bioavailability. Liposome encapsulation not only greatly improves the bioavailability of lotus leaf alkaloids but also reduces the dosage required.

[0099] 2) In the preparation of liposome lipid membranes, this lotus leaf alkaloid composition uses β-sitosterol instead of traditional cholesterol, which not only maintains the stability of the lipid membrane but also reduces cholesterol absorption, thus helping to enhance the weight loss effect of lotus leaf alkaloids.

[0100] 3) The lotus leaf alkaloid composition uses sodium alginate to perform secondary encapsulation of lotus leaf alkaloid liposomes, further improving the bioavailability of lotus leaf alkaloids. On the one hand, because lotus leaf alkaloids are positively charged lipophilic compounds, a large amount of lotus leaf alkaloids can be embedded and adsorbed onto the outer membrane of the liposomes without successful encapsulation, leading to easy loss. Encapsulation with sodium alginate reduces this loss. On the other hand, gastric juice has a destructive effect on lotus leaf alkaloids. Encapsulation with sodium alginate allows the lotus leaf alkaloid liposomes to exert an anti-acid effect, reducing the destruction of lotus leaf alkaloids by gastric juice, thus increasing the amount of lotus leaf alkaloids entering the intestine and improving its bioavailability.

[0101] 4) This lotus leaf alkaloid composition is economical, easy to operate, and has no side effects. It achieves the effect of weight loss treatment, avoiding the disadvantages of traditional methods that borrow from human methods to prevent and treat obesity in pet dogs, such as high cost and unsatisfactory results.

[0102] Not limited to this, any variations or substitutions conceived without inventive effort should be included within the scope of protection of this invention. Therefore, the scope of protection of this invention should be determined by the scope defined in the claims.

Claims

1. A lotus leaf alkaloid composition for preventing and treating obesity in pet dogs, characterized in that, The lotus leaf alkaloid composition for preventing and treating obesity in pet dogs is made from the following raw materials in parts by weight: 5-10 parts lotus leaf alkaloid, 50-100 parts egg yolk lecithin, 10-20 parts β-sitosterol, and 10-20 parts sodium alginate. The weight ratio of egg yolk lecithin to β-sitosterol is 5:1 to 8:

1. The weight ratio of the sum of egg yolk lecithin and β-sitosterol to lotus leaf alkaloid is 8:1 to 14:

1. The method for preparing the lotus leaf alkaloid composition for preventing and treating obesity in pet dogs includes: Step S1: Take 5-10 parts of lotus leaf alkaloid, 50-100 parts of egg yolk lecithin and 10-20 parts of β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the concentration of egg yolk lecithin 5-10 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved. Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:1 to 1:

3. Then place it on a magnetic stirrer to hydrate for 30 min. Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark. Step S4: Take 10-20 parts of sodium alginate according to the mass fraction, dissolve it in PBS solution with pH 6.8-7.4 to prepare a 0.5 mg / ml sodium alginate solution; Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition. Step S6: Seal the lotus leaf alkaloid composition obtained in step S5 and store it at 4°C in the dark.

2. The lotus leaf alkaloid composition for preventing and treating obesity in pet dogs according to claim 1, characterized in that, The weight ratio of egg yolk lecithin to β-sitosterol is 5.7:

1. The weight ratio of the sum of egg yolk lecithin and β-sitosterol to lotus leaf alkaloid is 12:

1.

3. A method for preparing a lotus leaf alkaloid composition for preventing and treating obesity in pet dogs, comprising: Step S1: Take 5-10 parts of lotus leaf alkaloid, 50-100 parts of egg yolk lecithin and 10-20 parts of β-sitosterol according to the mass ratio, add them to anhydrous ethanol solution to make the concentration of egg yolk lecithin 5-10 mg / ml, and sonicate at 200W under a water bath temperature of 40℃ until completely dissolved. Step S2: Inject the mixture solution obtained in step S1 into physiological saline that has been preheated to 40°C using a constant flow pump. The volume ratio of the mixture solution to physiological saline is 1:1 to 1:

3. Then place it on a magnetic stirrer to hydrate for 30 min. Step S3: Place the mixture suspension obtained in step S2 on a rotary evaporator to remove anhydrous ethanol, then sonicate at 200W for 30 min in an ice bath, filter through a 0.22μm filter membrane to initially obtain lotus leaf alkaloid liposomes, seal and store at 4℃ in the dark. Step S4: Take 10-20 parts of sodium alginate according to the mass fraction, dissolve it in PBS solution with pH 6.8-7.4 to prepare a 0.5 mg / ml sodium alginate solution; Step S5: Take the lotus leaf alkaloid liposomes obtained in step S3 and add them dropwise to an equal volume of sodium alginate solution obtained in step S4 while stirring. After the addition is complete, continue stirring for 1 hour to obtain the lotus leaf alkaloid composition. Step S6: Seal the lotus leaf alkaloid composition obtained in step S5 and store it at 4°C in the dark. in, The weight ratio of egg yolk lecithin to β-sitosterol is 5:1 to 8:

1. The weight ratio of the sum of egg yolk lecithin and β-sitosterol to lotus leaf alkaloid is 8:1 to 14:

1.

4. The application of the lotus leaf alkaloid composition for preventing and treating obesity in pet dogs according to claim 1 in the preparation of feed with the effect of preventing and treating obesity in pet dogs.

5. The application of the lotus leaf alkaloid composition for preventing and treating obesity in pet dogs prepared by the preparation method according to claim 3 in the preparation of feed with the effect of preventing and treating obesity in pet dogs.

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