Mixed extraction of noni and stevia for treatment of heart failure

Abstract

New use of combination of extraction of Noni and stevia reveals improvement effects on heart failure in animals.

Description

[0001] New use of mixed extraction of Noni and stevia for treatment of heart failure

[0002] Background

[0003] Chronic heart failure (CHF) is a significant global public health issue, Heart failure affects an estimated 64 million people globally, and making it a pandemiclevel health issue. The prevalence is expected to increase due to the aging population.

[0004] There is no specific pharmacological treatment for patients with heart failure with reduced ejection fraction (HFrEF) since no substantial prospective randomized clinical trial has been demonstrated exclusively in such a population. In addition, the incidence of Heart Failure with Preserved Ejection Fraction (HFpEF) is probably increasing due to the lack of an established effective treatment. Despite the important role of the Renin-Angiotensin-Aldosterone System (RAAS) in heart failure pathophysiology, RAAS blockers were found ineffective for HFpEF patients.

[0005] A therapeutic alternative may be represented by drugs stimulating the nonclassic RAAS (ACE2) as well as other emerging drug classes (such as SGLT2 inhibitors). However, no clinic trail results so far have revealed a significant improvement in either reduced ejection fraction (HFrEF) or preserved Ejection Fraction (HFpEF). Therefore, there is a gap or deficiency in the available medical options for treating CHF and represents a significant unmet medical need.

[0006] Noni (Morinda citrifolia) is a plant that has been traditionally used for its various health benefits. Noni juice may have a positive impact on cardiovascular health by exerting vasodilating effects on blood vessels, which can help regulate blood pressure and maintain a healthy heart. A study conducted at the University of Illinois College of Medicine at Rockford, USA, demonstrated that drinking noni juice can help reduce LDL (bad) cholesterol levels, total cholesterol content, homocysteine, and improve HDL (good) cholesterol levels in the body.

[0007] Studies found that Morinda citrifolia (Noni) can prevent inflammatory and oxidative stress, which may be factors in the development of heart failure. Noni aqueous extract (NAE) has been shown to ameliorate toxic effects caused by Bisphenol A (BP A), an endocrine disruptor, by increasing histone deacetylase (HD AC) expression and neovascularization activities, which can reduce cardiovascular risks.

[0008] While these findings suggest that noni may have potential benefits for heart health, the studies mentioned do not provide definitive evidence for the use of noni in treating heart failure.

[0009] In Stevia there is some research that suggests stevioside, a natural sweetener derived from the stevia plant, may have potential implications for heart health. There is limited and inconclusive evidence regarding the use of stevia or its components, such as stevioside, in the treatment of heart failure. A study analyzed the effect of Stevia (Stevia rebaudiana) in patients with chronic kidney disease (CKD) stages I to III. The study aimed to compare the effects of stevioside with conventional antihypertensive treatments. The study found that stevioside capsules (500 mg twice a day) showed good efficacy and tolerability in the long term in humans. However, the study's primary focus was on CKD patients, not specifically those with heart failure. Stevioside has been observed to have a significant decrease in systolic and diastolic blood pressure in patients with mild to moderate hypertension after one year of continued consumption of 750 mg / day and after two years of daily ingestion of 1500 mg / day

[0010] Stevioside has shown to inhibit the release of proinflammatory cytokines (TNF-a and IL-ip) and nitric oxide in a human monocytic THP1 cell line, suggesting potential anti-inflammatory effects. Stevioside and its related compounds may have antioxidant properties, which could be beneficial in managing oxidative stress related to various diseases

[0011] In conclusion, while stevia and its components like stevioside may have potential effects on blood pressure and other cardiovascular risk factors, there is no direct evidence from the provided search results to suggest that stevia is used for treating heart failure.

[0012] Briefly, neither Noni nor stevia alone showed no specific and significant effects in treatment of heart failure, while both Noni and stevia have been associated with potential cardiovascular benefits. However, there is no specific research on their combined use in treating heart failure so far.

[0013] In this invention we disclosed for the first time that treatment of heart failure by administration of a combination of both Noni extraction and stevia extraction. This combination of Noni extraction and stevia extraction is useful for treatment of cardiac hypertrophy and heart failure in TAC -induced long-term hypertrophy rats. It can also prevent cardiac remodeling by reducing the fibrosis and collagen deposition, and the size of cardiomyocytes and myocardium.

[0014] In addition, this invention disclosed that this mixture of combination of Noni extraction and stevia extraction ( mixture of noni and stevia) revealed significant therapeutically effects on heart failure with significant improvement in the symptoms: including fatigue, anorexia and pulmonary edema manifested by frequently cough and difficult in exercise with short breath in aged dogs.

[0015] Detail of Invention

[0016] This invention disclosed that a combination of mixture extraction of Noni and stevia reveals therapeutic effects on transverse aortic constriction (TAC)-induced cardiac hypertrophy and heart failure in animal. A significant increase in heart to body weight ratio (HW7BW), an index of cardiac hypertrophy, was observed in the 9-week TAC rats. The increase in the ratio of HW / BW was greatly reduced in TAC rats with treatment of mixture of Noni and stevia. The increased ratio of HW / BW was also accompanied by increased cardiomyocyte cross-sectional area which was increased by 76% percent in 9-week TAC rats comparing to sham rats. The increased myocardium hypertrophy and heart failure in TAC rats were significantly improved by treatment of mixture of Noni and stevia. This invention disclosed a new used of combination of Noni and Stevia in treatment of heart failure.

[0017] Noni powder preparing:

[0018] Preparation: start with high-quality, mature Noni fruits.

[0019] Drying: sliced to increase the surface area for efficient drying, drying can be done using methods including Natural Sunlight Drying which involves spreading the sliced Noni fruits in a single layer and exposing them to sunlight for several days until they are completely dried, and Dehydrator Drying: using a food dehydrator can speed up the process and provide more control over the drying conditions, such as temperature and air circulation.

[0020] Cleaning: to remove any dust, debris, or foreign particles that may have accumulated during the drying process.

[0021] Grinding: ground into a fine powder using a grinder or a mill. It should be done carefully to avoid overheating the product in order to preserve the nutritional and phytochemical properties of Noni.

[0022] The extraction of Noni powder:

[0023] Solvent: prepare the extraction solvent by mixing 80-50 parts of water with 20-50 parts of alcohol. Other solvent may be used for

[0024] acidification by adding acids such as citric acid, acetic acid, or lactic acid etc. in order to lower the pH level to compliance the food regulation.

[0025] Extraction: Immerse the Noni powder in the water-alcohol mixture. The ratio of Noni powder to solvent should be determined based on the desired concentration of the final extract. Allow the mixture to macerate for a period of time to allow the solvent to dissolve the bioactive compounds. This can take several hours or even days, depending on the specific protocol.

[0026] Filtration: after the maceration period, filter the mixture to separate the liquid extract from the solid Noni residue.

[0027] Concentration: the liquid extract can then be concentrated to remove the solvent and increase the potency of the bioactive compounds. This can be done through evaporation, often under reduced pressure and controlled temperature to prevent degradation of heat sensitive compounds.

[0028] Drying: Once the solvent has been removed, the remaining extract can be dried further to create a powder.

[0029] Preparation of stevia powder

[0030] The preparation of stevia powder involves extracting the sweet compounds known as stevioside from the leaves of the Stevia rebaudiana plant including:

[0031] Harvesting: the stevia leaves are harvested from the plant.

[0032] Drying: the leaves are then dried to reduce their moisture content which can be done through natural sun drying or using controlled drying equipment to ensure uniform drying and prevent degradation.

[0033] Crushing: the dried leaves are crushed or ground into a fine powder. Solvent Extraction: the similar process as noted above for Noni powder.

[0034] This invention also disclosed a co-extraction of Noni and stevia powders. Both Noni and stevia are mixed in proportions of 20:80 or 80:20. The solvents used and extraction process are similar to Noni as noted above.

[0035] Preparation of capsule:

[0036] After the extractions of both Noni and Stevia in their dried and powdered forms, Noni and Stevia were used to mixture at different proportions for encapsulation (typically 50:50). A size of capsules contains varies from 100 to lOOOmg of mixture of Noni and Stevia for different purpose of experiments.

[0037] Examples:

[0038] Example 1 Therapeutic effects on heart failure by administering the mixture of Noni and Stevia

[0039] Experiment materials

[0040] Animal: Adult male 30 Wistar rats, weighing 200g±20 g. Each rat was housed in an individual cage under standard conditions, and received standard laboratory diet ad libitum. Administration of the mixture powder of mixture of Noni and Stevia give orally at dosage: 30 mg / kg.

[0041] Experimental Methods

[0042] Establishment of cardiac hypertrophy (TAC transverse aortic constriction), a commonly used heart failure animal model and experimental protocol

[0043] TAC between the innominate artery and the left carotid artery was conducted to induce pressure overload. After surgery allowes animal to recovery. Sham control animals underwent the same operation, but without aortic constriction. All surgical procedures were performed with anesthetized with 3% pentobarbital intraperitoneally (i.p. 40mg / kg). Rats with TAC were randomly divided into three groups (n=8-10 rats) including a Sham control, TAC model, and mixture of both Noni and Stevia. (50:50) ( 30 mg / kg / d). Sham controls and TAC model were all treated with vehicle.

[0044] The TAC model was developed about 2 weeks after surgery. All treatment was given twice a day after two weeks surgery and the treatment last for additional 7 weeks (TAC for total 9 weeks). The animals were examined at 9 weeks at the end of the observation periods and after hemodynamic measurement in vivo, all animals were sacrificed. The hearts were removed and cut into different part for either nitrogen frozen or reagent fixation for further histopathological examination and biochemical analysis.

[0045] Measurements of cardio-dynamic parameters

[0046] Heart hemodynamic Analysis was conducted by pressure-volume (PV) catheter. Rats were anesthetized and placed on a warming pad (37°C) underwent tracheostomy with ventilation. The right internal carotid was identified and ligated cranially. A four-electrode pressure-volume catheter (model SPR-838, Millar Instruments Inc.) was advanced via right carotid artery into left ventricle until stable PV loops were obtained. Preload reduction achieved by compression of the inferior vena cava and parallel conductance values were obtained by the injection of 40pl hypertonic saline into the right jugular vein. Through a longitudinal inguinal incision, a polyethylene arterial catheter (PE10) connected to a pressure transducer was inserted into the distal abdominal aorta via the femoral artery. The left ventricular function and in peripheral pressure in each animal were examined. Parameter including: left ventricular ejection fraction (LVEF); Fractional Shortening (FS %), Left Ventricular End-Diastolic Volume (Vsd); Left Ventricular End-Systolic Volume (Ves). Stroke Volume (SV), maximal left ventricular pressure rising rate (Positive) +dp / dt max. (which can reflex the systolic heart failure), and maximal left ventricular pressure declining rate and (Negative)-dp / dt max.( which can reflex the diastolic heart failure).

[0047] Data were recorded on separate channels of the PowerLab system.

[0048] Histology Study

[0049] Tissue sections of rat hearts were fixed in 10% neutral -buffered formalin, embedded in paraffin, cut into 3 mm serial sections, and then stained with hematoxylin and eosin (H&E), picrosirius red or phalloidin. Nikon system and Zeiss confocal microscope were used to capture digital images. Stained with H&E was to evaluate cell size, stained with picrosirius red (sigma CA) was to test fibrosis using standard procedure. The cross-sectional cell area and interstitial collagen fraction were determined using computer-assisted image analysis (Image-Pro Plus software). Serum Biomarkers for cardiac injury Enzymes:

[0050] Lactate Dehydrogenase (LDH), Creatine Kinase (CK), Cardiac Troponin I (cTnl) and Cardiac Troponin T (cTnT); B-Type Natriuretic Peptide (BNP)

[0051] Effects on hypertrophy and heart failure

[0052] Hypertrophy and fibrosis

[0053] Adult Wistar rats were subjected to TAC for 9 weeks and a significant increase in heart to body weight ratio (HW7BW), an index of cardiac hypertrophy, increase 35%, (P < 0.001), which was accompanied by significant increased cardiac cross-sectional area. The cardiac hypertrophy (HW7BW), were significantly ameliorated by mixture of Noni and Stevia (reduce 12%, P < 0.01). The increment of cardiomyocyte cross-sectional area was also significant reduced.

[0054] Table 1. Effects of mixture of Noni and stevia on heart weight

[0055] and body weight in TAC rats (n=8)

[0056] Sham TAC TAC+Noni / stevia

[0057] HW(g) 0.66 ±0.04 0.94±0.07*** 0.72±0.03##

[0058] BW(g) 270± 9.7 271±7.3 250.5±5.1 HW / BW(mg / g) 2.45 ±0.1 3.32 ±0.16*" 2.92±0.1##

[0059] TAC transverse aortic constriction, HW heart weight, BW Body weight. Fibrosis: to determine whether mixture of Noni / stevia attenuates TAC -induced cardiac fibrosis, heart tissues were stained with picrosirius red to detect interstitial collagen distribution in left ventricular. In 9-week TAC groups, TAC induced significant interstitial fibrosis (P<0.05).and collagen content increased 7.5-fold compared to from sham groups. Treatment with mixture of Noni and Stevia (30mg / kg / d) resulted in 80.8% reductions in interstitial fibrosis comparing 9-week TAC groups.

[0060] Serum Biomarkers:

[0061] TAC rats show significant increase in LDH, Creatine Kinase (CK), Cardiac Troponin I (cTnl) and Cardiac Troponin T (cTnT) and BNP comparing to sham control. These biomarkers were significantly ameliorated after treatment of mixture of Noni and Stevia.

[0062] Effects on Cardiac function and Hemodynamic changes after treatment

[0063] There is significant declining in cardiac function after 9 weeks TAC including: significant decreased cardiac contractility (dp / dtmax) by 46% from sham control to TAC and reduced ejection fraction from sham control 67% to TAC 42% which decreased by 37%. These clearly indicate a deteriorated condition of heart failure.

[0064] After treatment of Mixture Noni and Stevia, the deteriorated condition of the heart significantly improved from the heart failure. There is significant recovered systolic contractile (+dp / dt max)by 65% comparing to TAC, and the ejection fraction recovered by 31% from TAC.

[0065] In addition, after treatment of Mixture Noni and Stevia, there is significant recovery in rate of relaxation (-dp / dt max) from TAC by 39%. This indicates a therapeutic effect on diastolic heart failure, with the incidence of preserved ejection fraction (HFpEF).

[0066] This invention disclosed that treatments of combination of mixture Noni and stevia have therapeutic effects on heart failure with either reduced ejection Fraction (HFrEF) or Preserved Ejection Fraction (HFpEF).

[0067] Table 2. Effects of mixture Noni and stevia on cardio-dynamic parameters before and after treatment in TAC heart failure

[0068] TAC+

[0069] Parameter Sham TAC _

[0070] Nom+Stevia

[0071] +dp / dt11354±9566142±345 10110±420

[0072] (mmHg / s)

[0073] -dp / dti2911±2116 7233±303 10103±481

[0074] (mmHg / s)

[0075] EF (%) 67±3 42±4 55±3

[0076] Ves (pL) 93±4 203±16 112±18

[0077] Ved (pL) 180±9 241±11 191±19

[0078] TAC transverse aortic constriction, EF ejection fraction Ves: ventricular end systolic volume; Ved: ventricular end diastolic volume

[0079] (Positive) +dp / dt max, (Negative) -dp / dt max.

[0080] Example 2. Effects of mixture of Noni and Stevia on heart failure

[0081] This study involves in 5 pet dogs ages around 7-10years diagnosis with heart failure. Dogs suffered with weakness and fatigue, increased respiratory rate with short breath, coughing more than usual, even with coughing up foam or blood due to lung fluid congestion, difficulty in exercising and have trouble catching its breath, and change in tongue or gum color. A reduced activity level or lethargy, lose their appetite, which can lead to weight loss and muscle wasting. Diagnosis confirmed with Echocardiography (ECHO) which including increased the size of the heart chambers, the thickness of the heart walls. A reduced ejection fraction (EF) less than 50 % and decreased Fractional Shortening (FS).

[0082] Once the dog diagnosed, the treatment with mixture of Noni and Stevia started under the monitoring by each individual care-taker of dog. The cuspules were given twice a day. Each cuspule contains lOOmg. The feeding of cuspules according the weight of each dog. Per day, the weight under 5kg give 2 cuspules, between 5 to 10 Kg give 4 cuspules and between 10 to 15 Kg give 8 cuspules. The treatment is for period 6-8 weeks for each dog. Assessments on each dog made at the beginning and the end of the treatment with mixture of Noni and Stevia.

[0083] This invention disclosed that the Echocardiography (ECHO) indicates significant improvements in the function of deteriorated heart in each treated dog with significantly increased ejection fraction (EF) and Fractional Shortening (FS) in comparison of the beginning of treatment. The size of heart and left ventricular wall thickness are both reduced or improved than the beginning of treatment.

[0084] According to the reports of caretakers of each dog, the dogs are eating well, energetic and much more exercise without difficult or short breach after treatments. Cough were seldom occurred comparing beginning treatments.

[0085] This invention disclosed that the mixture of Noni and stevia is effective in treatment heart failure in dogs. Furtherm ore, this invention disclosed that mixture of Noni and Stevia can be used to treat either reduced ejection fraction (HFrEF) or preserved Ejection Fraction (HFpEF).

[0086] Claims

[0087] What is claimed is:

[0088] 1, A method for treating cardiac hypertrophy and heart failure comprise administering combination of mixture of extraction from Noni and stevia to a human or animal in need.

[0089] 2, The method in accordance with claim 1, wherein the said heart failure is characterized by reduced ejection fraction (HFrEF) or preserved ejection Fraction (HFpEF).

Claims

Title: New use of mixed extraction of Noni and stevia for treatment of heart failureClaimsList of Claims1, A method for treating cardiac hypertrophy and heart failure comprise administering combination of mixture of extraction from Noni and stevia to a human o animal in need.2, The method in accordance with claim 1, wherein the said heart failure is characterized by reduced ejection fraction (HFrEF) or preserved ejection Fraction (HFpEF).