DESC:FIELD
The present disclosure relates to the field of homeopathy based formulation.
DEFINITIONS
As used in the present disclosure, the following terms are generally intended to have the meaning as set forth below, except to the extent that the context in which they are used indicate otherwise.
Nosode: The term “nosode” refers to a homeopathic preparations of organic material derived from inactivated disease products, cultures of microorganisms (e.g. bacteria, fungi and viruses) or parasites, infected or pathologically changed material or decomposition products from humans or animals, rendered safe during the homeopathic manufacturing process.
Decimal Potency: The term “decimal potency” refers to the principle that the first potency should contain one-tenth part of the original drug and each succeeding potency should contain one-tenth part of the potency preceding it. The decimal potency is denoted by suffixing the letter 'X' or 'D' to the number indicating the potency, i.e. the first potency is 1X or 1D in the decimal scale, followed by 2X or 2D and so on.
Centesimal Potency: The term “centesimal potency” refers to the principle that the first potency must contain one-hundredth part of the original drug and each succeeding potency must contain one-hundredth part of the preceding one. This scale is designated by suffixing the letter 'C' or 'CH' to the number indicating the potency, i.e., the first potency is 1C or 1CH, followed by 2C or 2CH and so on.
Potentization: The term “potentization” refers to the process of making a remedy more potent by serial dilution and succussion. Potentization is the process for the reduction, according to scale, of crude, inert or poisonous medical substances to a state of physical solubility, physiological assimilability and therapeutic activity and harmlessness, for use as Homeopathic healing remedies.
Trituration: The term “trituration” refers to grinding one compound into another to dilute one of the ingredients, add volume for processing and handling, or to mask undesirable qualities.
Succussion: The term “succussion” refers to the vigorous shaking of a diluted homeopathic preparation in order to activate the medicinal substance. Succussion is the process of potentization, by which preparation of medicine takes place by the use of a liquid vehicle like alcohol or water, by shaking in a definite method.
Potency: The term “potency” refers to the power that is derived by the grades of medicinal power developed by the process of dynamization. Potency is a result of a series of successive dilutions, according to scale and friction through succussion or trituration.
Parasitemia: The term “parasitemia” refers to the quantitative content of parasites in the blood. It is used as a measurement of parasite load in an organism and an indication of the degree of an active parasitic infection.
BACKGROUND
Malaria is a dominant parasitic infection caused by a parasite of the genus Plasmodium. Plasmodium falciparum and Plasmodium vivax are the two major species which are infective to humans. Plasmodium falciparum is responsible for 85 % of the malaria cases in the world. Two other plasmodial species, Plasmodium ovale and Plasmodium malariae, occur with irregularity, but the latter may induce considerable immunopathology, particularly in children.
Considerable research has been carried out for the treatment of malaria in various fields of medicine, such as, Allopathy, Ayurveda, Homeopathy and the like. Despite years of research and major expenditures for the control of disease, malaria remains a major world-wide public health problem.
In recent years, there has been a dramatic increase in the use of non-allopathic compositions and therapeutic regimens for malaria, due to the drug-resistance developed by the malarial parasites. Use of alternative medical approaches and medicines, such as homeopathy, is gaining widespread acceptance.
Homeopathy is based on the doctrine that “likes cures like” and makes use of minute dose to cure a disease. Homeopathy provides safe and long lasting results by energizing the body's curative powers.
Therefore, there still remains scope to develop effective malarial nosode to combat infection by malarial parasites.
OBJECTS
Some of the objects of the present disclosure, which at least one embodiment herein satisfies, are as follows:
It is an object of the present disclosure to ameliorate one or more problems of the prior art or to at least provide a useful alternative.
Another object of the present disclosure is to provide a homeopathy based formulation.
Still another object of the present disclosure is to provide a homeopathy based formulation which is effective against malaria.
Other objects and advantages of the present disclosure will be more apparent from the following description, which is not intended to limit the scope of the present disclosure.
SUMMARY
The present disclosure relates to a homeopathy based formulation. The formulation comprises at least one potentized mixture of malaria nosode, and a vehicle. The ratio of the potentized mixture of malaria nosode to the vehicle can be 1:9 for 1X potency and 1:99 for 1C potency. Typically, the malaria nosode can be obtained from a malaria parasite selected from the group consisting of Plasmodium falciparum Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, and combinations thereof. The potentized mixture of malaria nosode is typically obtained by serial dilution and succussion of the malaria parasite in the vehicle in the range of 1C to 1 million C. The vehicle can be selected from the group consisting of alcohol, water and sachhrum lactose.
The present disclosure also provides a process for preparing a homeopathy based formulation. The process comprises preparing a malaria nosode, which is potentisized to obtain a potentized mixture. The step of potentisizing the malaria nosode is then iterated 30 to 10,000 times to obtain the homeopathy based formulation of the present disclosure. The potency of the formulation can be in the range of 30C to 10,000C potency. The malaria nosode can be prepared by cultivating a malaria parasite on a suitable growth medium and isolating the malaria parasite from the growth medium to obtain isolated malaria parasite. The isolated malaria parasite is titurated with a vehicle to obtain the malaria nosode. Typically, the ratio of the isolated malaria parasite to the vehicle can be in the range of 1:99 to 50:50.
The homeopathy based formulation of the present disclosure can be used for the treatment of malaria
DETAILED DESCRIPTION
Malaria is a dominant parasitic infection caused by a parasite of the genus Plasmodium. Plasmodium falciparum and Plasmodium vivax are the two major species which are infective to humans. Plasmodium falciparum is responsible for 85 % of the malaria cases in the world.
Plasmodium falciparum is a protozoan parasite, belonging to the Plasmodium species, which causes malaria in humans. Plasmodium falciparum is transmitted by the female Anopheles mosquito. Plasmodium falciparum is the causative agent of the malignant or falciparum malaria, having the highest complication rates and mortality. Globally, malaria is one of the most prevalent parasitic disease of humans and claims the lives of more children worldwide than any other infectious disease.
Plasmodium falciparum can cause severe malaria as it can multiply rapidly in the blood, and can thus cause severe blood loss (anemia). In addition, the infected parasites can clog the small blood vessels in the infected individual. Cerebral malaria is caused, when the brain of an individual is infected with Plasmodium falciparum, which can be fatal.
Plasmodium vivax is a protozoan parasite and a human pathogen. It is the most frequent and widely distributed cause of recurring (Benign tertian) malaria. It is less virulent than Plasmodium falciparum, however, vivax malaria can lead to severe disease and death due to splenomegaly (a pathologically enlarged spleen).
Plasmodium ovale is a species of parasitic protozoa that causes tertian malaria in humans. It is less dangerous as compared to Plasmodium falciparum.
Plasmodium malariae is a parasitic protozoa that causes malaria in humans. Though, it is present worldwide, it is not as dangerous as compared to P. falciparum or P. vivax. It causes fevers that recur at approximately three-day intervals (a quartan fever), longer than the two-day (tertian) intervals of the other malarial parasites, hence its alternative names are quartan fever and quartan malaria. In recent years, there has been a dramatic increase in the use of non-allopathic compositions and therapeutic regimens for malaria, due to the drug-resistance developed by malarial parasites. The use of alternative medical approaches and medicines, such as homeopathy, is gaining widespread acceptance.
Homeopathy is a system of alternative medicine developed in 1796 by Samuel Hahnemann based on his doctrine of like cures like (similia similibus curentur): a claim that a substance that causes the symptoms of a disease in healthy people would cure similar symptoms in sick people. Homeopathy is based on the principles of:
i. like cures like: if the symptoms of your cold are similar to poisoning by mercury, then mercury would be your homeopathic remedy;
ii. minimal dose: the remedy is taken in an extremely dilute form; typically, one part of the remedy to around 1,000,000,000,000 parts of water; and
iii. the single remedy: no matter how many symptoms are experienced, only one remedy is taken, and that remedy will be aimed at all those symptoms.
Nosodes are broad-spectrum, widely used homoeopathic preparations sourced from biological materials, such as, cultures or clinical samples of microorganisms (e.g. bacteria, fungi, and viruses) or from parasites, diseased tissues (cancerous tissues), or decomposition products from humans or animals. Nosodes are applied for treatment as well as for alleviating the symptoms of acute as well as chronic diseases. The Homoeopathy Pharmacopeia of India (HPI) had divided the source material into four groups N-I, N-II, N-III, and N-IV, depending on the nature of the source material, whether organisms are capable of producing endotoxins, exotoxins, made from purified toxins or made from microbes, viruses, or clinical material from diseased subjects.
The present disclosure envisages a homeopathy based formulation that is effective for the treatment of malaria. The nosode obtained from malaria parasite is diluted to an extent that no molecules of the original substance remain in the homeopathy based formulation.
The digestion of hemoglobin in the food vacuole of the malarial parasite produces high quantities of redox active toxic free heme. The Plasmodium species detoxifies the free heme by the formation of hemozoin (beta-hematin). Hence, the present disclosure targets the hemozoin formation to develop the homeopathy based formulation that is effective in the treatment of malaria.
In an aspect of the present disclosure, there is provided a homeopathy based formulation. The formulation comprises at least one potentized mixture of malaria nosode, and a vehicle. The ratio of the potentized mixture of malaria nosode to the vehicle can be 1:9 for 1X potency and 1:99 for 1C potency. In an embodiment, the potentized mixture of malaria nosode is obtained by serial dilution and succussion of the malaria parasite in the vehicle in the range of 1C to 1 million C.
Typically, the malaria nosode is prepared from a malaria parasite selected from the group consisting of Plasmodium falciparum Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, and combinations thereof.
The vehicle can be selected from the group consisting of alcohol, water and sachhrum lactose.
In another aspect of the present disclosure, there is provided a process for preparing a homeopathy based formulation. The process comprises preparing a malaria nosode, and then potentisizing the malaria nosode to obtain the homeopathy based formulation of the present disclosure. The process is described hereinafter in detail.

Initially, the malaria nosode is prepared by cultivating a malaria parasite on a growth medium. The malaria nosode can be prepared from any known strain of malaria parasite. In accordance with the embodiments of the present disclosure, the malaria parasite is selected from the group consisting of Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, and combinations thereof. The malaria nosode is grown on a suitable growth medium, as per known in-vitro cultivation parasite protocols. In an embodiment, the malaria parasite is cultivated on a growth medium comprising Red Blood Cells (RBCs).
The malaria parasite is then isolated from the growth medium using known techniques. In one embodiment, the malaria parasite is isolated by subjecting the RBCs infected with the malaria parasite to centrifugation. Typically, centrifugation of the RBCs can be carried out at a speed in the range of 1000 rpm to 2000 rpm for a time period in the range of 5 minutes to 60 minutes, at a temperature in the range of 10 °C to 50 °C. Further, the RBCs can be lysed using a suitable lysing agent. In one embodiment, the lysing agent is saponin.
The malaria parasite can be isolated from the centrifuged and lysed RBCs. The number of malaria parasites can be calculated based on % parasitemia and the total volume of packed blood cells.
The isolated malaria parasite is then used as the original stock for making the malaria nosode. A pre-determined number of isolated malaria parasite is triturated with a vehicle to obtain the malaria nosode. Typically, the ratio of isolated malaria parasite to the vehicle can be in the range of 1:99 to 50:50. The malaria nosode is potentisized using the vehicle to obtain a potentisized mixture. The step of potentisizing the malaria nosode is iterated 30 to 10,000 times, to obtain the homeopathy based formulation of the present disclosure. Typically, the potency of the homeopathy based formulation is in the range of 30C to 10,000C.
The vehicle can be selected from the group consisting of alcohol, water and sachhrum lactose. In an embodiment, the vehicle for the tituration step is Saccharum lactis. In another embodiment, the vehicle for the potentisization step is selected from alcohol, and water. Typically, the potentization can be performed using an electro-mechanical potentizer.
Typically, the safety of nosode in various potencies can be established as per the sterility testing mentioned in the HPI. Further, the presence and growth of the source organism(s) in various potencies is established and documented. Additional testing, such as DNA test, pro-viral DNA test, ultra-electron microscopy, Reverse Transcription Polymerase Chain Reaction (RT-PCR) method with positive and negative control can be carried out to establish the safety of the nosode for human use.
The prepared nosode can be lyophilized, so that nosode having the same characteristics can be reproduced. All the preparations are carried out in stringent biosafety compliant environment with minimum handling, using sealed containers and disposable auto-tip pipettes. The safety regarding the oral use of the nosodes is established through pathogenetic trials in healthy volunteers and clinical trials.
Typically, the homeopathic formulation is prepared in a liquid vehicle and may thereafter be prepared in various delivery forms, including liquid delivery forms, solid delivery forms such as tablets or other solid carriers on which the liquid is deposited, and other oral delivery forms, including time release formulations
The malaria nosodes envisaged in accordance with the present disclosure exhibit anti-malarial activity and can be used in the treatment of malaria.
The present disclosure is further described in light of the following laboratory scale experiments which are set forth for illustration purpose only and not to be construed for limiting the scope of the disclosure. These laboratory scale experiments can be scaled up to industrial/commercial scale and the results obtained can be extrapolated to industrial/commercial scale.
Experimental details:
Experiment-1: Preparation of the homeopathy based formulation in accordance with the present disclosure
Plasmodium falciparum 3D7 (procured from Biosciences and Bio-Engineering department of IIT-B (Mumbai)) was used to prepare the malaria nosode. The parasite Plasmodium falciparum 3D7 is suggested to have originated from Africa. Plasmodium falciparum was cultivated on a growth medium comprising Red Blood Cells (RBCs). The RBCs (O +ve) for cultivating Plasmodium falciparum was collected from healthy volunteers using an anti-coagulant, 10 % citrate-phosphate-dextrose-adenine (CPDA).
1.8 x108 parasites (isolated Plasmodium falciparum) were titurated with 9 parts of Saccharum lactis powder to obtain a malaria nosode mixture having 1X potency.
The malaria nosode was further diluted and succussed by taking one part of the 1X potency and 9 parts of Saccharum lactis powder to obtain Plasmodium falciparum nosode having 2X potency. The 2X malaria nosode was then potentisized (electro-mechanical potentizer having a 12,099 Nm force) using alcohol to obtain the homeopathy based formulation having a potency of 30C. A 1:99 ratio of 2X malaria nosode to alcohol was used to obtain the homeopathy based formulation having the potency of 30C.
Experiment-2: Evaluation of the antimalarial activity of the homeopathy based formulation prepared in accordance with the present disclosure
All the chemicals except Hydrochloric acid (HCl) were purchased from Sigma-Aldrich Chemical Company, while HCl was purchased from Merck.
Homeopathy based formulation obtained in Experiment-1 (Sample-1) was used to evaluate the antimalarial activity, alcohol and potentized alcohol (potentized up to 30C potency) were used as the vehicle control, and known anti-malarial drug Chloroquine diphosphate (500 µg) was used as the standard.
All the samples were incubated with 3 mM of hematin, 10mM oleic acid, and 1M HCl. The final volume was adjusted to 1mL using sodium acetate buffer, pH 5. The reaction mixtures were incubated overnight at 37 °C with constant gentle shaking. After incubation, samples were centrifuged (14,000 rpm, 10 min, at 21 °C) and the hemozoin pellet obtained was repeatedly washed with incubation (15 min at 37 °C with regular shaking) in 2.5 % (w/v) SDS in phosphate buffered saline followed by a final wash in 0.1M sodium bicarbonate until the supernatant was clear (6 washes). After the final wash, the supernatant was removed and the pellets were dissolved in 1mL of 0.1M NaOH before determining the hemozoin content by measuring the absorbance at 400 nm. The results were recorded as % inhibition (I %) of heme crystallization compared to vehicle controls using the following equation:
??% = [(AN-AS)/AN] × 100
Where AN: Absorbance of vehicle control and AS is absorbance of test samples
The results obtained are summarized in Table-1. The results are presented as mean ± SD of three independent experiments carried out in duplicates. Statistical significance between the groups was assessed by one-way analysis of variance (ANOVA) followed by a post hoc test. The accepted level of significance for the test was P < 0.05. All tests were carried out using GraphPad Software (Version 3.06).

Table-1: Effect of the samples on hemozoin formation
S. No. Sample % Inhibition
1. Alcohol 44.72 ± 3.01
2. Potentized alcohol 43.64 ± 2.48
3. Sample-1 57.96 ± 8.52
4. Chloroquine diphosphate (500 µg) 53.83 ± 8.33
All values represent Mean ± SD;
@ p<0.05; @@ p<0.01 as compared to the Untreated Control group;
(ANOVA followed by post-hoc tests)
ANOVA is a collection of statistical models used to analyze the differences among group means and their associated procedures (such as "variation" among and between groups). S.D. represents the standard deviation. Post hoc test consists of looking at the data—after the experiment has concluded—for patterns that were not specified a priori. Graphpad is scientific 2D graphing and statistics software.
It is clearly seen from Table-1 that there is a significant inhibition of the hemozoin formation by the homeopathy based formulation of the present disclosure (Sample-1) as compared to alcohol, and potentized alcohol as vehicle controls. Further, the inhibition is also higher than the inhibition exhibited by the standard, Chloroquine diphosphate.
Similar homeopathy based formulations were prepared using Plasmodium malariae, Plasmodium ovale, Plasmodium vivax. The homeopathy based formulations prepared from Plasmodium malariae, Plasmodium ovale, and Plasmodium vivax exhibited a similar antimalarial activity as that exhibited by homeopathy based formulation prepared using Plasmodium falciparum nosode.
TECHNICAL ADVANCEMENTS
The present disclosure described herein above has several technical advantages including, but not limited to, the realization of
? homeopathy based formulation, that can be used for the treatment of malaria; and
? an effective and economical homeopathy based formulation which can be used in combination with other medicines.
The embodiments as described herein above, and various features and advantageous details thereof are explained with reference to the non-limiting embodiments in the description. Descriptions of well-known aspects, components and molecular biology techniques are omitted so as to not unnecessarily obscure the embodiments herein.
The foregoing description of specific embodiments so fully reveals the general nature of the embodiments herein, that others can, by applying current knowledge, readily modify and/or adapt for various applications of such specific embodiments without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments herein have been described in terms of preferred embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the spirit and scope of the embodiments as described herein. Further, it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.
Having described and illustrated the principles of the present disclosure with reference to the described embodiments, it will be recognized that the described embodiments can be modified in arrangement and detail without departing from the scope of such principles.
While considerable emphasis has been placed herein on the particular features of this disclosure, it will be appreciated that various modifications can be made, and that many changes can be made in the preferred embodiment without departing from the principles of the disclosure. These and other modifications in the nature of the disclosure or the preferred embodiments will be apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.
,CLAIMS:WE CLAIM:
1. A homeopathy based formulation comprising a homogenized mixture of:
? at least one potentized mixture of malaria nosode; and
? a vehicle;
wherein said malaria nosode is obtained from at least one malaria parasite selected from the group consisting of Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, and combinations thereof; and the ratio of said potentized mixture of malaria nosode to said vehicle is 1:9 for 1X potency and 1:99 for 1C potency.
2. The formulation as claimed in claim 1, wherein said vehicle is selected from the group consisting of alcohol, water, and sachhrum lactose.
3. The formulation as claimed in claim 1, wherein said potentized mixture of malaria nosode is obtained by serial dilution and succussion of said parasite in said vehicle in the range of 1C to 1 million C.
4. A process for preparing a homeopathy based formulation, said process comprising the steps of:
a. preparing a malaria nosode by:
• cultivating at least one malaria parasite on a suitable growth medium;
• isolating said malaria parasite from said growth medium to obtain isolated malaria parasite;
• titurating said isolated malaria parasite with a vehicle to obtain to obtain said malaria nosode, wherein the ratio of said isolated malaria parasite to said vehicle is in the range of 1:99 to 50:50;
wherein said malaria parasite is at least one selected from the group consisting of Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, and combinations thereof.
b. potentisizing said malaria nosode using said vehicle to obtain a potentized mixture;
c. iterating said step (b) for 30 to 10,000 times, to obtain said homeopathy based formulation;
wherein the potency of said formulation is in the range of 30C to 10,000C potency.
5. The process as claimed in claim 4, wherein said vehicle is selected from the group consisting of alcohol, water, and sachhrum lactose.
6. The process as claimed in claim 4, wherein said vehicle for said tituration in step (a) is sachhrum lactose.
7. The process as claimed in claim 4, wherein said vehicle for said potentisization in step (b) is selected from alcohol, and water.
8. The process as claimed in claim 4, wherein said potentization is performed using an electro-mechanical potentizer.
9. The homeopathy based formulation as claimed in claim 1 for use in the treatment of malaria.