Application of whey protein in preparation of medicine for resisting epilepsy-related depression

Abstract

The invention provides application of whey protein in preparation of a medicine for resisting epilepsy-related depression, and belongs to the technical field of nutritional neuroimmunology. According to the application of the whey protein in preparation of the product for resisting epilepsy-related depression, the whey protein oral preparation can improve depression symptoms, avoid occurrence and aggravation of epilepsy symptoms, improve intestinal microenvironment, avoid common digestive tract reactions of SSRIs and enhance compliance of patients at the same time.

Description

Technical Field

[0001] The present invention relates to the technical field of nutritional neuroimmunology, and in particular to use of whey protein in preparing a drug for treating epilepsy-related depression. Background Art

[0002] Accurately assessing the severity of depressive symptoms is a key step in developing treatment strategies. The Beck Depression Inventory (BDI) is a widely used assessment tool in clinical practice. This 21-item scale assesses a patient's mood, cognition, and physical symptoms, with each item scored from 0 to 3 points, for a total score ranging from 0 to 63. Based on the BDI score, patients can be categorized as follows: mild depression: 14-19 points; moderate depression: 20-28 points; severe depression: ≥29 points.

[0003] Based on the above classification, treatment usually follows the following principles:

[0004] 1. For mild depressive episodes in patients with epilepsy, psychological interventions (such as cognitive behavioral therapy) are considered first-line treatment. If medication is necessary, selective serotonin reuptake inhibitors (SSRIs) are the preferred medication, with a Grade B recommendation. Commonly used SSRIs include sertraline, paroxetine, and fluoxetine.

[0005] 2. For moderate to severe depressive episodes, SSRIs remain the preferred medication (Grade B). However, for patients who do not respond adequately to first-line treatment, switching to norepinephrine-serotonin reuptake inhibitors (SNRIs) such as venlafaxine is considered a reasonable alternative (Grade C).

[0006] 3. Antidepressant drug treatment should be maintained for at least 6 months after the first depressive episode is relieved; if the patient has a history of previous episodes, it is recommended to extend it to 9 months; for those with severe depression or residual symptoms, the treatment time should even be extended until the symptoms completely subside.

[0007] Although SSRIs have a good antidepressant effect in epilepsy-related depression, their potential impact on epileptic seizures remains controversial. Some studies have shown that some SSRIs may aggravate epilepsy symptoms in some cases, especially when used in high doses or for a long time. For example:

[0008] 1. Clinical research evidence: In a prospective study, sertraline treatment of depression in epileptic patients resulted in an increase in seizure frequency in approximately 6% of patients, suggesting a potential epileptogenic effect. Furthermore, another retrospective observational study also found that SSRIs may increase seizure frequency in some patients.

[0009] 2. Animal Experimental Studies: The epileptogenic effects of SSRIs are more pronounced in animal models. For example, in an epileptic mouse model established using the amygdala kindling method, continuous microinfusion of fluoxetine for 30 days significantly accelerated the onset of epilepsy and increased the severity of seizures. Another study showed that in a pentylenetetrazol (PTZ)-induced epilepsy model, high-dose venlafaxine exhibited significant epileptogenic effects, manifested by shortened seizure latency, increased seizure intensity, and increased mortality.

[0010] In addition to the potential risk of worsening epilepsy, SSRIs themselves carry a range of side effects that shouldn't be ignored. According to the U.S. Food and Drug Administration (FDA)'s adverse reaction monitoring data for 10,782 patients treated with fluoxetine, approximately 5% of patients reported adverse reactions of varying degrees, primarily digestive symptoms, including nausea, vomiting, diarrhea, and loss of appetite.

[0011] In summary, current treatment strategies for epilepsy-related depression primarily rely on antidepressant medication interventions based on the BDI score. However, some medications may induce or worsen epileptic seizures under certain conditions, and their side effects can affect patient compliance, limiting their clinical application. Therefore, the development of safer and more effective alternative treatment options has become an urgent and pressing issue. Summary of the Invention

[0012] This invention aims to use whey protein in the preparation of a drug for treating epilepsy-related depression. This drug is intended for use as an adjunctive intervention for epilepsy-related depression, demonstrating multiple biological benefits, including neuroprotection, antidepressant effects, antioxidant activity, and regulation of the gut-brain axis. The drug can be developed as an oral powder, liquid formulation, or medical food, suitable for the long-term management of epilepsy patients.

[0013] The technical solution of the present invention is achieved as follows:

[0014] The present invention provides use of whey protein in preparing a product for resisting epilepsy-related depression.

[0015] As a further improvement of the present invention, the product is a tablet, capsule, liquid preparation, or powder.

[0016] As a further improvement of the present invention, the liquid preparation is a solution, a suspension, or an injection.

[0017] As a further improvement of the present invention, the product is a medicine or a medical food.

[0018] As a further improvement of the present invention, the whey protein is used alone.

[0019] As a further improvement of the present invention, the whey protein is used as an auxiliary treatment means in combination with an anti-epileptic drug.

[0020] Implementation principle:

[0021] 1. Biological effects of amino acids rich in whey protein

[0022] Whey protein is rich in tryptophan, a precursor for the neurotransmitter serotonin (5-HT). Animal studies have shown that whey protein can enhance 5-HT synthesis, regulate brain 5-HT levels, and improve the mood of tumor-bearing mice. Whey protein is also rich in cystine, a key substrate for glutathione synthesis. Glutathione is an important antioxidant in the brain that scavenges free radicals and reduces oxidative stress, thus playing a key role in neuroprotection.

[0023] 2. The neurotropic properties of whey protein can effectively affect the brain

[0024] Lactoferrin, a compound in whey protein, has neurotropic properties. Lactoferrin can bind to specific receptors on brain vascular endothelial cells, enabling targeted delivery across the blood-brain barrier (BBB). Studies have found that lactoferrin has highly expressed receptors in neurons and brain microvessels, demonstrating antioxidant, anti-inflammatory, and neurotrophic factor-promoting effects in models of neurodegenerative diseases such as Parkinson's disease.

[0025] 3. Regulate intestinal flora and influence brain activity through the brain-gut axis

[0026] Whey protein can also influence gut-brain signaling by regulating the composition of the intestinal flora. The intestinal microbiome synthesizes metabolites such as short-chain fatty acids and neurotransmitter precursors, regulating central nervous system function through the vagus nerve, immune system, and endocrine pathways. Whey protein has been shown to improve intestinal flora diversity and enhance intestinal barrier function, thereby indirectly improving mood and cognitive status.

[0027] The present invention has the following beneficial effects: the oral whey protein preparation can improve depressive symptoms while avoiding the occurrence and aggravation of epilepsy symptoms, while also improving the intestinal microenvironment, avoiding common gastrointestinal reactions of SSRIs, and enhancing patient compliance. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0029] Figure 1 Figure 2 shows the weight change of mice (left) and the Racine score of epilepsy (right);

[0030] Figure 2 This is the result of the open field experiment;

[0031] Figure 3 This is the result of the elevated plus maze experiment;

[0032] Figure 4 Results of the forced swimming test (left) and the tail suspension test (right) are shown. DETAILED DESCRIPTION

[0033] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0034] Example 1

[0035] 1. The structural composition of whey protein:

[0036] ①Material composition: Whey protein is composed of approximately 70.2% β-lactoglobulin, 14.3% α-lactalbumin, 8.6% bovine serum albumin and approximately 6.9% immunoglobulins5.

[0037] Nutritional composition: Whey protein typically contains 93% protein (on a dry basis). It also contains 0.1% lactose, 1.3% fat, approximately 4.7% water, and 2.7% ash. Each 100g of WPI provides approximately 375 kcal and 23mg of cholesterol. WPI is rich in minerals, including sodium (805mg / 100g), calcium (120mg / 100g), potassium (120mg / 100g), phosphorus (180mg / 100g), magnesium (15mg / 100g), and trace amounts of iron and chloride, demonstrating its superiority as a low-fat, high-protein nutritional supplement.

[0038] ③Amino Acid Composition: WPI is rich in essential and functional amino acids. Branched-chain amino acids include leucine (10.6g / 100g), isoleucine (6.2g / 100g), and valine (5.2g / 100g). Essential amino acids include tryptophan (1.7g / 100g), lysine (9.7g / 100g), methionine (2.4g / 100g), phenylalanine (2.7g / 100g), serine (4.1g / 100g), and threonine (6.4g / 100g). WPI is also rich in glutamic acid (17.1g / 100g), aspartic acid (9.4g / 100g), and lysine (9.7g / 100g).

[0039] 2. Application method:

[0040] ① Oral powder or liquid preparation, 1-2 times a day for 4-8 weeks;

[0041] ②It can be used in epilepsy model animals or humans as an auxiliary intervention method.

[0042] 3. Implementation Case (Animal Experiment)

[0043] In a PTZ-induced epilepsy mouse model, the whey protein composition was administered for 4 weeks. The results showed that:

[0044] ① Behavioral scores (including elevated maze test, open field test, tail suspension test, and forced swim test) were significantly improved;

[0045] ② There was no significant increase in the frequency of epileptic seizures, and the safety was good.

[0046] Example 2

[0047] (1) Experimental plan

[0048] 1. Animal grouping: A total of 48 mice were divided into a control group (Con), a model group (PTZ), a fluoxetine treatment group (FLX), and a whey protein treatment group (WPI), with 12 mice in each group. The PTZ, WPI, and FLX groups will use epilepsy mouse models.

[0049] 2. Construction of epilepsy mouse model: An epilepsy model was established by intraperitoneal injection of pentylenetetrazol (PTZ) once every other day.

[0050] 3. Drug Administration: For the whey protein-treated mice, whey protein was dissolved in saline and administered daily by gavage at 35 mg / kg. For the fluoxetine-treated group, the drug was also dissolved in saline and administered daily by gavage at 2 mg / kg. The control group received the same volume of saline.

[0051] 4. Behavioral Testing: After 10 PTZ injections, mice were subjected to behavioral testing. Testing included the tail suspension test, open field test, forced swim test, and elevated maze test. All behavioral experiments strictly adhered to animal ethics and were conducted according to the principle of increasing stress.

[0052] (2) Experimental results

[0053] 1. General

[0054] The mice used in this batch (2025-05 to 2025-06) of experiments were of the C57BL / 6J strain, all male, totaling 20, purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. The mice were raised at the Experimental Animal Center of Tsinghua University. The breeding environment was strictly controlled to simulate the natural circadian rhythm, with a regular light and dark cycle, and standard feed and drinking water were provided free access. All experimental operations followed the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health (NIH) of the United States and were approved by the Laboratory Animal Ethics Committee of the School of Medicine of Tsinghua University (approval number: 23-GXH1.G23-2). During the experiment, two mice in the model group died of epileptic seizures, and one mouse in the whey protein intervention group also died. The remaining animals completed the experimental process smoothly.

[0055] 2. Whey protein intake can effectively alleviate weight loss and the severity of epileptic seizures in epileptic mice:

[0056] To evaluate the effect of whey protein on the overall health of epileptic mice, the body weight of the mice was recorded every 4 days during the experiment. The results showed that from the 16th day, the body weight of the mice in the epilepsy model group (PTZ group, n=5) began to be significantly lower than that of the control group (Con group, n=5), which were 20.198g and 21.982g respectively (p<0.05). By the end of the 28th day, the body weight of the mice in the whey protein intervention group (WPI group, n=4) was significantly higher than that in the model group (n=3), which were 23.77g and 19.283g respectively (p<0.05), indicating that whey protein can effectively alleviate epilepsy-induced weight loss. It is worth noting that there was no significant difference between the WPI group and the fluoxetine treatment group (FLX group, n=5, average body weight 23.724g) ( Figure 1 Left), showing that whey protein is as effective as classic antidepressants in improving body weight.

[0057] In addition, to evaluate the effect of whey protein on the severity of epileptic seizures, the epileptic behavior of mice was observed after each PTZ injection and graded according to the Racine score: Grade 1 is mouth and face twitching, Grade 2 is head twitching, Grade 3 is forelimb lifting and twitching, Grade 4 is falling to the ground with limb twitching, and Grade 5 is limb rigidity with strong whole-body twitching. After the 10th PTZ injection, the average Racine score of mice in the model group (n=3) was 5.0, indicating severe epileptic seizures; while the average score of mice in the WPI group (n=4) was 3.5, which was significantly lower than that of the model group (p<0.05), indicating that whey protein intervention can effectively reduce the intensity of epileptic seizures ( Figure 1 right)

[0058] 3. Behavioral experimental results showed that whey protein significantly improved the depressive-like behavior of mice

[0059] In the open field experiment, the roadmap results ( Figure 2 A) shows that epilepsy model mice (PTZ group) exhibited typical depressive-like behavior, and the frequency of entering the central area was significantly reduced. Both whey protein (WPI group) and fluoxetine (FLX group) interventions could reverse this behavioral pattern. Statistical analysis results showed that the number of times entering the central area in the PTZ group (n=3, average 3.67 times) was significantly reduced compared with the control group (Con group, n=5, average 12.6 times) (p<0.01); while the WPI group (n=4, average 8.5 times, p=0.0763) and the FLX group (n=5, average 8.8 times, p<0.05) were significantly increased compared with the PTZ group ( Figure 2 B). In terms of the time spent in the central area, the PTZ group (average 7.05 seconds) was significantly shorter than the Con group (average 50.05 seconds, p<0.01), while the WPI group (average 29.01 seconds, p<0.05) and the FLX group (average 30.31 seconds, p<0.05) were significantly longer than the PTZ group ( Figure 2 C). It is noteworthy that there was no significant difference in total distance moved between the groups ( Figure 2 D), suggesting that the behavioral changes are not due to differences in motor ability.

[0060] In the elevated plus maze test, the heat map results ( Figure 3 A) shows that mice in the PTZ group tended to stay in the closed arm, and the frequency and time of entering the open arm were significantly reduced, showing anxiety and depression-like behaviors. Specifically, the number of times the PTZ group entered the open arm (n=3, average 1.33 times) was significantly lower than that of the Con group (n=5, average 8.2 times, p<0.001); while the WPI group (n=4, average 4.5 times, p<0.05) and the FLX group (n=5, average 6.8 times, p<0.01) were significantly higher than those in the PTZ group ( Figure 3B). In terms of the open arm residence time, the PTZ group (average 3.91 seconds) was significantly lower than the Con group (average 46.5 seconds, p<0.01), while the WPI group (average 18.25 seconds, p<0.05) and the FLX group (average 24.74 seconds, p<0.05) were significantly improved ( Figure 3 C).

[0061] In the forced swim test and tail suspension test, the immobility time of mice in the PTZ group was significantly prolonged, further supporting the manifestation of their depressive-like behavior. In the forced swim test, the average immobility time of the PTZ group (n=5) was 149.71 seconds, which was significantly longer than that of the Con group (average 27.11 seconds, p<0.01); while the WPI group (n=5, average 95.83 seconds) and the FLX group (n=5, average 57.21 seconds) both had significantly shorter immobility times (p<0.01) ( Figure 4 Left). In the tail suspension test, the average immobility time of the PTZ group was 206.37 seconds, which was also significantly higher than that of the Con group (average 58.36 seconds, p<0.01); the WPI group (average 134.86 seconds) and the FLX group (average 113.10 seconds) both showed significant improvement (p<0.01) ( Figure 4 right).

[0062] In summary, whey protein intervention showed antidepressant effects in multiple behavioral experiments, suggesting that it has potential neuromodulatory effects in the intervention of epilepsy-related depression.

[0063] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. Use of whey protein in preparing a product for treating epilepsy-related depression.

2. The use according to claim 1, characterized in that The products are tablets, capsules, liquid preparations, and powders.

3. The oral use according to claim 2, characterized in that The liquid preparation is a solution, a suspension, or an injection.

4. The use according to claim 1, characterized in that The product is a medicine or a medical food.

5. The use according to claim 1, characterized in that The whey protein was used alone.

6. The use according to claim 1, characterized in that The whey protein is used as an auxiliary treatment means in combination with an anti-epileptic drug.

Images