WO2021182647A1

WO2021182647A1 - Fortification of foods with carica papaya seeds for treatment of schistosomiasis

Info

Publication number: WO2021182647A1
Application number: PCT/KE2020/050003
Authority: WO
Inventors: Elijah SONGOK
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-03-11
Filing date: 2020-10-09
Publication date: 2021-09-16
Anticipated expiration: 2022-09-11
Also published as: WO2021182647A4

Abstract

Schistosomiasis is classified as among the world's neglected tropical diseases (NTD). Morbidity to S. mansoni is greatest among school-age children who typically have the highest burden of infection. In 2001, World Health Organization (WHO) passed a resolution for large-scale mass drug administration (MDA) using chemotherapy to deworm vulnerable children through school-based programs. However, chemotherapeutic drugs are not recommended for children under the age of 6 years due to their toxicity, despite the fact that this is the age group most infected with schistosomiasis. Additionally, the current school-based MDA does not consider child malnutrition a very common malady in African countries. The current invention discloses the making of a food composition comprising common edible cereals like maize, sorghum or a combination, fortified with extracts from Carica papaya (C. papaya) fruit seeds to form a mixture, and the use of this mixture in the management and or treatment of schistosomiasis.

Description

TITLE: FORTIFICATION OF FOODS WITH CARICA PAPAYA SEEDS FOR TREATMENT OF SCHISTOSOMIASIS

Field of invention: The invention pertains to health and preventive medicine, more particularly, to a treatment provided through fortification of foods.

Background Art:

Schistosomiasis, also known as Bilharzia is a tropical disease caused by a trematode parasite of the genus Schistosoma (Colley et al., 2014). Infections by intestinal parasites -Ascaris lumbricoides, taenia spp ( hookworm ) Trichuris trichura, and Schistosoma mansoni ( S.mansoni ) are an important public health concern (WHO, 1998). These maladies are most prevalent in poor settings of the developing world, with the highest burden of infection registered among school-aged children (Booth et al., 1992). Intestinal parasites are commonly associated with malnutrition, anemia, impaired growth, poor school attendance, and impaired cognition (Hotez et al., 2007). In 2001, the World Health Assembly passed a resolution for endemic countries to implement large-scale mass drug administration (MDA) of school-aged children using albendazole chemotherapy for Ascaris, Taenia and Trichuris (soil-transmitted helminths) and praziquantel (PZQ) for S. mansoni (Hall et al., 2008;

WHO, 2006). While MDA has significantly contributed to reducing the burden of these infections, several concerns still exist over the large-scale use of chemotherapeutic drugs in deworming. The large population of children and the high frequency of dosage may pose a challenge to the sustainability of these programs. Further, the MDAs exert increasing drug pressure on parasite populations, a circumstance that is likely to favor parasite genotypes that can resist chemotherapy. For example, the existence of S. mansoni tolerance to PZQ has been reported in Senegal, where the parasitological cure rate 12 weeks after treatment was as low as 18% (Albonico et al., 2008). In regions in Egypt and Kenya where there has been heavy exposure to PZQ, there are emerging reports of S. mansoni and S. haematobium resistance to treatment (Brindley and Sher, 1987; Ross et al., 2002). Besides, the MDA mechanism is only limited to school- aged and pre-school children, while the other community population is neglected (Masaku et al., 2017).

Additionally, PZQ is bitter tasting and therefore, not administered to newborn and young children. But, perhaps the most significant drawback to chemotherapeutic MDA, is its inability to consider child malnutrition, a widespread problem in most resource limited countries (Ismail et al., 1996; Stephenson et al., 2000), this is exacerbated by the fact that the PZQ drugs have to be taken with meals- a rare phenomenon in resource-limited settings rural schools. Consequently, there has been an urgent need to design treatment alternatives that are not only effective, affordable, and sustainable but also directly enhances child nutrition status. Quite a number of natural foods contain active properties that have been shown to act against schistosomiasis, for example, ole-gum-resin (myrrh) an extract from Commiphora molmol has been used in treatment of Schistosoma haematobium, (EP080492) , elsewhere, W02015123480 and US20200009099 discloses the method of treatment and prevention of schistosomiasis using polyunsaturated fatty acid (PUFA).

On the other hand, the application of papaya in the treatment of ailments has been described in various reports. For example, Choi, 2015 (CA2946095A1) describes a method of preparation made from oat flour and papaya fruit, used in treatment of skin-related ailments such as eczema and dry skin, further, Choi, 2002, (652353499T3), describes a method of using Carica papaya for treatment of digestive disorders, in addition, US20080069907A1 discloses the use of extracts of papaya from various parts of papaya plant including leaves for the treatment of multiple forms of cancer such as stomach cancer. Moreover, papaya has for centuries been used as an antihelminthic compound in India (Global burden of disease collaboration, 2016; Krishnakumar, 1960), Central and South America (Dar et al., 1965) and throughout the world (Werner et al., 1992). In Cuba, papaya seed extract has been formulated to capsules and dispensed at a dosage of lg three times a day for helminthic treatment (Volpato et al., 2009), while BR 102013027442 A2 described a method of preparing nanoparticles from papaya latex for antihelmintic properties. Laboratory studies have confirmed that various preparations of papaya seeds can kill helminths and S. mansoni cercaria effectively in vitro, and in infected animals (Mokua, 2013; Muchika et al., 2010). In Africa, the clinical importance of papaya seed extract as an antihelminthic was confirmed in a study from Nigeria.

In a randomized trial to determine the effectiveness of dried papaya seeds on human intestinal parasitoses, 60 asymptomatic children with microscopic stool evidence of intestinal parasites who received a lg honey- emulsified dose of papaya seed, showed a parasite clearance rate of 77% after seven days as compared to 17% clearance rate on those who were given honey alone (z = 4.40, p = .0000109) (Okeniyi et al., 20007). No adverse effects were observed from both groups of children. Follow up phytochemical analysis has shown that the bio-active antihelminthic ingredient in papaya seeds was benzyl isothiocyanate, a chemopreventive compound with antioxidant properties (Kermanshai et al., 2001).

Summary of the invention

We have discovered that use of cereal porridge fortified with Carica papaya seed powder can be used in management or treatment of schistosomiasis disease, caused by S. mansoni. Clinical studies showed that papaya seed fortified porridge had a significant reduction in the prevalence of schistosomiasis in infected subjects. Therefore in accordance with this invention, Carica papaya seed powder is effective against schistosomiasis prevalence. More specifically it relates to the use of food compositions fortified with Carica papaya seed powder for the treatment or management of schistosomiasis.

In particular embodiment, a method is provided for making Carica papaya seed powder for fortification of cereal flour, comprising the steps of obtaining the Carica papaya fruit, getting the seeds from the fruit and drying the Carica papaya seeds, washing the dried seeds in glacial acetic acid; rinsing off the glacial acetic acid from the seeds with tap water for at least 4 times; re-drying the seeds for at least 1 hour and milling the Carica papaya seed into powder.

In another embodiment, the Carica papaya seeds are washed in 8% glacial acetic acid. In another embodiment of the invention, the Carica papaya seeds are washed in glacial acetic acid for at least 30 minutes.

In yet another embodiment, the Carica papaya seeds are dried in the sun or the dryer. In another embodiment of the invention, the Carica papaya seeds are milled to powder using a gauge 16 sieve.

In another embodiment, we provide a method of fortifying cereal flour food composition with Carica papaya seed powder.

In another embodiment of the invention, the fortified cereal flour food composition comprises about 5 parts of cereal flour to 1 part of the Carica papaya seed powder.

In another embodiment a micronutrient premix supplied nutrients are added to Carica papaya seed fortified cereal flour food composition.

In another embodiment the cereal flour comprises, maize, sorghum, millet, rice, wheat, or mixtures thereof. In another embodiment, the composition can be made into the form of porridge, biscuits, powder, flour or tablet.

In another embodiment, a method is provided for use of food composition fortified with Carica papaya seed powder for treating or managing schistosomiasis, comprising the steps of identifying a subject infected with schistosomiasis diseases, making porridge from cereal flour fortified with Carica papaya seed powder and administering to the subject an effective dose of the Carica papaya porridge.

In another embodiment, we provide a method of making biscuits fortified with Carica papaya seed powder. Comprising steps of mixing sugar and margarine in a mixing bowl and beating the mixture at medium speed till the mixture is fluffy white. Adding to this composition, flour and baking powder and mixing for at least 2 minutes, thereafter adding milk or water and a preferred essence then mixing the contents to obtain a consistency; removing the mixture from the mixing bowl and piping the mixture onto a greased baking sheet using desired nozzle shapes and baking the biscuit mixture in an oven at 180 °C for at least 25 minutes.

In another embodiment, the biscuits fortified with Carica papaya seed powder are used for the treatment of schistosomasis. Detailed disclosure of the invention:

The objective of this invention was to provide a food composition, which is fortified with Carica papaya seed powder for the treatment or management of schistosomiasis in human subjects As a food source, with no known harmful effects, enough evidence suggests that papaya seeds can be a sustainable candidate for deworming school children. Its reduction to practice through fortifications to common foods and healing has not been demonstrated. Accordingly, the present invention relates to a fortification strategy using Carica papaya fruit seeds to common edible cereals like maize, sorghum, millet, rice, wheat, or any other combination as mass drug administration tool against S. mansoni and other intestinal parasites in human subjects. Porridge made from maize (corn) flour is one of the most prevalent traditional school meal snacks in developing countries. Because of its low cost and popularity, it has been adopted as a component in school meals and is often prepared and given as a snack at break time and forms as breakfast in many communities. Similarly, biscuits are a delicacy among school-going children and equally affordable. The method has an advantage that, the papaya seeds are a natural product with no additives or preservatives, and high heat stability. The seeds are also readily available and affordable; furthermore, the method of treatment of schistosomiasis proposed by this invention, is not only an affordable treatment option for schistosomiasis but also provides the patient with other nutritional benefits and at the same time a filling meal for the school going children especially in resource-poor countries with adverse food scarcity. With the knowledge of papaya seed as an anthelminthic, the current invention discloses a method of preparing a cereal deworming porridge, as follows.

Carica papaya seed is extracted from whole ripe fruit; they are then washed with tap water and dried thoroughly under the sun. The dried seeds are again re-washed in 8% glacial acetic acid in a ratio of 5:8 (seeds: glacial acetic acid). The seeds and glacial acetic acid preparation are appropriately mixed and allowed to stand for 30 minutes. The seeds are then rinsed with tap water 3 to 4 times to wash off all the glacial acetic acid; they are then placed on trays and dried in a dryer. The dry seeds are then taken out from the drier. The seeds are then milled into powder, thereafter, one part of the papaya seed powder is mixed with 5 parts of the cereal flour. The Carica papaya seed fortified flour is then divided into small manageable portions of preferably 10kg portions, to each of these portions, a micronutrient premix is added in the ratio of 1:400 (one part micronutrient premix to 400 parts Carica papaya seed fortified flour) and mixed well. The fortified flour is then packaged and stored in a cool, dry place. The components of the micronutrient premix are as described in table 1 below.

Table 1. Components of the micronutrient premix

Vitamin A (IU) 4,162,500 Iron (mg) 40,000

Zinc (mg) 60,000 Thiamine (mg) 8000 Riboflavin (mg) 7000 Niacin (mg) 61,000 Vitamin B6 (mg)' 10,000 Folate (mg) 3000 Vitamin B12 (pg) 14,000

The Carica papaya seed fortified porridge, or otherwise referred to as the deworming porridge for the treatment of schistosomiasis is prepared as described below; The deworming porridge is prepared by heating water to boil; the premix prepared above is then added to the boiled water in a ratio of 1:9, respectively, followed by stirring to mix, while heating. The porridge is then allowed to cook for approximately 15-30 minutes. The treatment of schistosomiasis infected patients using the fortified porridge is as follows;

Each infected schistosomiasis patient is given a capful (300ml) of the deworming porridge once a day for 30 days, which is sufficient to cure the disease. The current invention also describes a method of preparing Carica papaya seed fortified biscuits for the treatment of schistosomiasis.

The biscuit consists of the following components: the papaya seed flour (2.25%), wheat flour (42.70%), sugar (12.15%), icing sugar (6.07%), margarine (0.10%), baking powder (0.25%), vanilla essence (or any kind of flavoring agent) (0.24%), water (or milk) (12.15%) and eggs (0.05%). The biscuits are prepared as follows:

Where the sugar and margarine are mixed in a mixing bowl and beat at medium speed till the mixture is fluffy white. Wheat flour, Carica papaya seed powder and baking powder are added and mixed for 2 minutes, thereafter; milk or water and a preferred essence (vanilla, strawberry and etc) are added and mixed to obtain a consistency. The mixture is then removed from the mixing bowl and piped onto greased baking sheet using desired nozzle shapes. Finally the mixture is baked in an oven at 180°C for a time of between 20-25 minutes or thereof

EXAMPLES

The following example is meant to illustrate the methods and materials of the present invention and in no way intended to limit the invention in any way. EXAMPLE 1. STUDY DESIGN

The preliminary evaluation of the effect of the deworming porridge on 5. mansoni was carried out on children in schools of Kirinyaga County, central region of Kenya. Four schools in Mwea Division of Kirinyaga County of Kenya with a high prevalence of S. mansoni were chosen for a clinical trial. The Carica papaya seed fortified flour was given to the schools to prepare the porridge for all children in Early Childhood Development (ECD) classes. The porridge was prepared as follows;

270 ml of water was boiled, then 30g of maize flour fortified with Carica papaya seed powder, and micronutrients were added and stirred to mix well while heating. The porridge was then allowed to cook for 15-30 minutes.

Prior to porridge feeding, all children had their stool samples taken for 5. mansoni microscopy (Kato-Katz) after which each child received 300 ml deworming porridge every school day for 25 days. A follow-up stool sample was taken at the end of the follow-up period. Schools were then closed for the 1 month August holidays without taking the papaya seed fortified porridge, and on reopening schools, a stool sample was again taken from the children for analysis.

Statistical analysis Data were analyzed using SPSS version 13.0 software (SPSS, Inc., Chicago, IL). The parasite infection prevalence, defined as the percentage of children with eggs in faeces was calculated at baseline and at follow up. Parasite eggs counted are expressed as eggs per gram (epg) of faeces. The initial parasite load was corrected for by including it in the analysis of variance. Only individuals for whom there were complete data on infection status from baseline to 25 days were included in this analysis.

Results

Baseline characteristics

A total of 269 children participated in the trial (140 female vs 130 male). The mean age was 5 years (+SD1.0; range 3- 11); and mean weight 17.2 kg (±SD 2.5 range 10-28.4) (Table 2). The overall prevalence of 5. mansoni at baseline was 37.1 % (100 of 270).

Effects of treatment on parasite infestation

The papaya seed fortified porridge reduced the prevalence of S. mansoni to 19.3% (52 of 270, p <0.001) ((Fig. 1) after 25 days of use. The prevalence, however, rebounded to 27.03% (73 of 270) (Fig. 1) after the school holidays.

Table 3 and figure 2 shows the distribution of intensities of infections, expressed as epg of faeces, before treatment and after follow up 1 and 2. Geometric mean intensity was significantly reduced (p < p<0.0001) from 82.2 to 17.3 epg during the 25 day intervention period in the group that consumed papaya seed fortified porridge but rebounded to 73.9 epg during follow up 2 after the school holidays (Fig. 2, Table 3). Table 2. Demographics of participants at baseline- by gender

N (observations) Variable N mean SD

Age 133 4.9 0.87

Height 140 105 8.19

Female 140 Weight 140 16.7 2.15

Age 122 5.2 1.14

Height 129 108 8.21

Male 129 Weight 129 17.6 2.81

Figure 1 Prevalence of Schistosoma mansoni infection before start of fortified porridge feeding (Baseline), after fortified porridge (Followup 1) and after break (Follow-up 2).

Table 3 Mean egg counts among all study participants

N Mean egg count SD Min

Baseline 269 82.2 316 ΊG

Follow up 1 269 17.3 71.2 o

Follow up 2 269 73.9 302 o Figure 2 Mean egg count of Shistosoma masoni among all study participants before (Baseline), after (Follow-up 1) fortified porridge feeding and after break (Follow-up 2).

This study proves that enhancing cereal flour with the seeds of Carica papaya can be used as a treatment alternative for S. mansoni infection and substituting this in the school feeding program, may act as an alternative or supplement to current national school based deworming programs in Africa especially to child populations that are out of reach to current chemotherapeutic approaches. As a simple school meal, the papaya seeds fortified flour porridge brings the added benefit in the fight against child malnutrition. Besides, Papaya seeds are used traditionally as a natural deworming agent. Several studies have also confirmed its antiparasitic potency. In most communities, papa seeds are discarded and their use in this innovation provides a cost-effective and environmentally friendly strategy for the treatment of schistosomiasis. Among several advantages achieved by the present invention, therefore, may be noted the provision of a method for preparing Carica papaya seed powder; the provision of a new cereal flour containing a food composition which can serve as a vehicle for treatment or management of schistosomiasis and nutrient provision; the provision of a new biscuit containing a food composition which can serve as a vehicle for treatment or management of schistosomiasis; the provision of a method for treatment or management of schistosomiasis in schistosomiasis infected subjects.

Claims

Claims:

1.) A method for making Carica papaya seed powder for fortification of cereal, wherein the Carica papaya seed powder so prepared is used for treatment or management of schistosomiasis. 2.) A method of claim 1, wherein making of the Carica papaya seed powder comprises, acquiring ripe Carica papaya fruits, obtaining seeds from the Carica papaya, drying the Carica papaya seeds, washing the dried seeds in glacial acetic acid; rinsing off the glacial acetic acid from the seeds with tap water for at least 4 times; re-drying the seeds for at least 1 hour and milling the Carica papaya seed into powder.

3.) A method of claim 2, wherein the Carica papaya seeds are washed in

8% glacial acetic acid.

4.) The method according the claim 2 and 3, wherein the Carica papaya seeds are washed in glacial acetic acid for at least 30 minutes.

5.) The method according to claim 2, wherein the Carica papaya seeds are dried in the sun or the dryer.

6.) A method according to claim 2 wherein the Carica papaya seeds are milled to powder using a gauge 16 sieve. 7.) A method of making a cereal flour food composition fortified with

Carica papaya seed powder

8.) Method according to claim 7 wherein, the Carica papaya seed fortified food composition comprises from about 5 parts of cereal flour to 1 part of the Carica papaya seed powder. .) A method according to claim 8 wherein a micronutrient premix supplied nutrients is added to the Carica papaya seed fortified cereal flour in the ratio of 1 part micronutrient premix to 400 parts Carica papaya seed fortified cereal flour.

10.) Method according to claim 7 and 8 wherein the cereal flour comprises, maize, sorghum, millet, rice, wheat, or mixtures thereof. 11.) The food composition according to claim 7 to 10, wherein the food composition is in the form of porridge, powder, flour or tablet.

12.) The use of food composition fortified with Carica papaya seed powder prepared according to any of the methods in claim 7 to 11 for treating or managing schistosomiasis, the use comprising administering to a subject in need thereof an effective dose of the food composition.

13.) A method of making biscuits fortified with Carica papaya seed powder.

14.) A method according claim 13, the method comprising mixing sugar and margarine in a mixing bowl and beating the mixture at medium speed till the mixture is fluffy white. Adding to this composition flour and baking powder and mixing this for at least 2 minutes, thereafter adding milk or water and an essence mixing the contents to obtain a consistency; removing the mixture from the mixing bowl and piping the mixture onto a greased baking sheet using desired nozzle shapes and baking the biscuit mixture in an oven at

180°C for at least 25 minutes. The use of Carica papaya seed fortified biscuits for treating or managing schistosomiasis.