Formulation for targeting cancer in humans and canines using venom

Abstract

The present invention provides formulation/composition comprising phytonutrients with natural chlorotoxins for targeting cancer, infection, inflammation and pain without any side effects and a method for synthesizing the same. The raw materials are cleaned and dried and to prepare coarse powder of 40 mesh size, extracted with solvent (comprising water:alcohol in a ratio of 40:60) in a ratio of 4:1 with overnight soaking. Prior to cold extraction, the mixture is macerated for 4 hours. The mixture is refluxed for 2 hours at 80 C. The addition of ethanol, maceration and refluxing steps are repeated three times and above solvent is added, if required. The residue is checked for complete extraction after every refluxing step. The extract/residue is filtered and concentrated under vacuum. The extract/residue is vacuum tray dried at 70-80 C. for 12 hours. The extract/residue is scraped and dried lumps of the extract/residue are milled. The extract/residue is sieved and packed.

Claims

1. A method for the synthesis of a nutraceutical supplement comprising phytonutrients and natural chlorotoxins for targeting disease comprising of: A. First creating a powder from said phytonutrients comprising of the steps: a. Forming a first phytonutrients powder from plants comprising said phytonutrients, wherein said plants are selected from the group consisting of Glycyrriza glabra, Piper longum, Picrorhhiza kurroa, Phyllanthus amarus, Bauhinia variegata, Terminalia chebulain, and combinations thereof; b. Applying a solvent to said first phytonutrients powder to form a first mixture; c. Macerating said first mixture; d. Separating the soluble compounds in said first mixture using cold extraction; e. Separating said soluble compounds from the said first mixture into a first residue and first soluble compounds; f. Applying a solvent to said first residue to form a second mixture; g. Macerating said second mixture; h. Separating the soluble compounds in said second mixture using cold extraction; i. Separating said soluble compounds from the said second mixture into a second residue and second soluble compounds; j. Applying a solvent to said second residue to form to form a third mixture; k. Macerating said third mixture; I. Separating the soluble compounds in said third mixture using cold extraction; m. Separating said soluble compounds from the said third mixture into third residue and soluble compounds; n. Concentrating said third residue by filtering and concentrated under vacuum to create an extract; o. Drying said extract; p. Milling said extract into a second phytonutrients powder; B. Second, extracting chlorotoxins from the venom of a scorpion comprising of: a. Securely holding the scorpion; b. Applying an electrical stimulus to said scorpion; c. Collecting the venom containing the chlorotoxins in a 1 ml vial; d. Freezing said venom; e. Taking the frozen venom and forming a supernatant using distilled water; f. Lyophilizing said supernatant to form a lyophilized chlorotoxins powder containing chlorotoxins; C. Third, adding 0.0001 grams of the lyophilized powder containing chlorotoxins to 1 gram of said phytonutrients powder to form a chlorotoxins and phytonutrients powder; D. Fourth, mixing said chlorotoxins and phytonutrients powder to form a uniform powder mixture of phytonutrients and chlorotoxins; E. Fifth, taking said uniform powder mixture of phytonutrients and chlorotoxins and mixing an effective amount thereof with at least one excipient to formulate said nutraceutical supplement.

2. The method of claim 1, wherein the disease is selected from the group consisting of cancer, Alzheimer's, heart disease, epilepsy, and pain management.

3. The method of claim 1, wherein the chlorotoxins are from venom extractions from a Blue Scorpion.

4. The method of claim 1, wherein the chlorotoxins are collected from the scorpion species selected from the group consisting of Bothriuridae, Buthidae, Caraboctonidae, Chactidae, Chaerilidae, Diplocentridae, Euscorpiidae, Gigantoscorpionidae, Heterometrus swammerdami, Hemiscorpiidae, Palaeopisthacanthidae, Pseudochactidae, Scorpionidae, Superstitioniidae, Typhlochactidae, Vaejovidae, African Androctonus australis, Hadrurus hirsutus, Albino Scorpions, Black Scorpions, Asian Forest Scorpions, Blue Scorpions, Emperor Scorpions, Deathstalker Scorpions, Lesser Brown Scorpions, Red Claw Scorpions, Red Scorpions, Sand Scorpions, Tailless Whip Scorpions, and Whip Scorpions.

5. The method of claim 1, wherein said excipient is selected from the group consisting of water, aqua, sorbitol, glycerin, sodium benzoate, sodium carboxymethylcellulose (Na-CMC), honey, sucralose and citric acid.

6. The method of claim 1, wherein one or more of said solvents comprises a water:alcohol ratio of 40:60.

7. The method of claim 1, further comprising forming the nutraceutical supplement into a physiological vehicle, wherein said physiological vehicle is selected from the group consisting of a pill, an injectable liquid, an oral liquid, an oral tablet, a topical cream, a nasal spray, and a dry powder aerosol.

8. The method of claim 7, wherein the said injectable liquid is formed by mixing the nutraceutical supplement with amounts of suspending liquid, stabilizers, and preservatives.

9. The method of claim 8, wherein the suspending liquid is selected from the group consisting of deionized water, saline, plasma, and interstitial fluid.

10. The method of claim 8, wherein the preservatives are dissolved in said suspending liquid and said preservatives are selected from the group consisting of benzoic acid, salts, sorbic acid, paraben substitute, grape fruit seed, benzyl alcohol, phenoxyethanol, sodium benzoate, potassium sorbate, and parabens.

11. The method of claim 8, wherein the stabilizers are dissolved in said suspending liquid and said stabilizers are selected from the group consisting of albumin, proteins, phenols, polyols, glycine, poloxamers, and cyclodextrins.

12. The method of claim 7, wherein the physiological vehicle is a pill which is formulated by mixing the nutraceutical supplement with amounts of adjuvants, excipients, stabilizers, and preservatives.

13. The method of claim 12, wherein said excipient is selected from the group consisting of water, aqua, sorbitol, glycerin, sodium benzoate, sodium carboxymethylcellulose (Na-CMC), honey, sucralose, and citric acid.

14. The method of claim 13, wherein said adjuvants are selected from the group consisting of aluminum phosphate, aluminum hydroxide, docosahexaenoic acid, and MF59.

15. The method of claim 7, wherein the oral liquid is formed by mixing the nutraceutical supplement with amounts of liquid, stabilizer, and preservatives.

16. The method of claim 15, wherein the liquid is selected from the group consisting of deionized water, saline, plasma, and interstitial fluid.

17. The method of claim 15, wherein the stabilizer is selected from the group consisting of polyols, albumins and other proteins, surfactants, poloxamers, cyclodextrins and the preservatives are selected from benzoic acid, salts, sorbic acid, paraben substitute, grape fruit seed, benzyl alcohol, phenoxyethanol, sodium benzoate, potassium sorbate and parabens.

18. The method of claim 15, wherein the preservatives are selected from the group consisting of methyl-, ethyl-, and propyl-parabens, sodium benzoate, benzoic acid, methyl paraben/sodium benzoate, benzyl alcohol, phenoxyethanol, grape fruit seed, potassium sorbate, and parabens.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) Exemplary embodiments of a system and method are shown in the accompanying drawings. The other objects, features, and advantages will occur to those skilled in the art from the following description of the preferred embodiment and the accompanying drawings in which:

(2) FIG. 1 illustrates a flow chart explaining a method for synthesizing the formulation/composition comprising phytonutrients and natural chlorotoxins for targeting cancer, according to one embodiment of the present invention.

(3) Although the specific features of the present invention are shown in some drawings and not in others. This is done for convenience only, as each feature may be combined with any or all of the other features in accordance with the present invention.

DETAILED DESCRIPTION OF INVENTION

(4) The Definition of scorpion includes all of the following Scorpion families which includes by definition: Bothriuridae, Buthidae, Caraboctonidae Chactidae, Chaerilidae, Diplocentridae, Euscorpiidae, Gigantoscorpionidae, Heterometrus swammerdami, Hemiscorpiidae, Palaeopisthacanthidae, Pseudochactidae, Scorpionidae, Superstitioniidae, Typhlochactidae and Vaejovidae, and all Seventy-two scorpion species, belonging to Hemiscorpiidae.

(5) In the following detailed description, reference is made to the accompanying drawings that form a part hereof, and in which the specific embodiments that may be practiced is shown by way of illustration. These embodiments are described in sufficient detail to enable those skilled in the art to practice the embodiments and it is to be understood that the logical, mechanical, and other changes may be made without departing from the scope of the embodiments. The following detailed description is therefore not to be taken in a limiting sense.

(6) The various embodiments of the present invention provide a formulation/composition comprising phytonutrients and natural chlorotoxins for targeting cancer in humans, felines and canines, and other pet animals without any side effects. The embodiments of the present invention also provide a method for the synthesis of a formulation comprising phytonutrients and natural chlorotoxins for targeting cancer.

(7) FIG. 1 illustrates a flow chart explaining a method for synthesizing the formulation/composition (RX002CxN) comprising phytonutrients and natural chlorotoxins such as Blue Scorpion Venom Chlorotoxin for targeting cancer, according to one embodiment of the present invention. The phytonutrients are selected from the group comprising of Glycyrrizaglabra, Piper longum, Picrorhiza kurroa, Phyllanthus amarus, Bauhinia variegate, and Terminalia chebula. The herbal raw material can also include Cannabis sativa (cannabinoids). The herbal raw materials are cleaned and dried and mixed together to form a uniform mixture. Then a coarse powder (101) is prepared such that it can pass through a 20-40 mesh size by either grinding or pulverizing using a suitable tool such as a mortar and pestle, or a processor such as a IKA Handheld Analytical Mill, or an IKA Continuous Feed Grinding machine EW-04300-30 with IKA 2836001 Continuous feed grinding drive. Then solvent comprising water/alcohol in a ratio 40:60 is added to the coarse powder mixture in a ratio 4-parts powder and 1-part solvent and the solvent is mixed thoroughly to obtain a homogeneous mixture (102). The mixture is macerated for at least 4 hours to 12 hours, followed by a cold extraction of 12 hours forming homogenous mixture (103). Cold extraction is achieved by taking the homogeneous mixture of solvent and phytonutrients powder and dissolving the mixture in warm water, and then rapidly cooling the mixture. The insoluble compounds precipitate out of the water, while the soluble ones stay dissolved. The solution can then be separated by reflux, filtration, or decantation. In the preferred method the mixture is refluxed (104) for 2 hours at 80 C. The addition of solvent maceration (105) and refluxing steps are repeated a minimum three times and sufficient quantity of solvent is added, if required. The residue is checked (106) for complete extraction after every refluxing step. The extract/residue is filtered and concentrated under vacuum (108). The extract/residue is vacuum tray dried at 70-80 C. for 12 hours (107). The extract/residue is scraped and the dried lumps of the extract/residue are milled (109). The natural chlorotoxin is added to the extract/residue and blended homogenously (110). The extract/residue is sieved and packed (111). The percentage yield is calculated according to the weight of the raw materials. The example utilizes the Blue Scorpion Venom Chlorotoxin venom however the opportunity to expand the viable chlorotoxin pool of compounds by utilizing any one of the more than 1004 known species of scorpion's in the treatment of disease's such as heart disease, cancer, Alzheimer's, epilepsy, inflammation and pain management is incorporate because any one of the vast number of scorpion's could be used as a donor animal and this specification includes the incorporation of any of these animals being used as an alternative sources of chlorotoxin's. For this example, the Blue Scorpion Venom Chlorotoxin venom is extracted by a mild electro-stimulation method that includes stimulating and scorpion restraining devices. However researchers have had success milking scorpions in the Buthidae family, the Scorpionidae family which includes the emperor scorpion (Pandinus imperator), and the Heterometrus swammerdami scorpion as well as specific species such as African Androctonus australis, Hadrurus hirsutus, Albino Scorpions, Black Scorpions or Asian Forest Scorpions, Blue Scorpions, Emperor Scorpions, Deathstalker Scorpions, Lesser Brown Scorpions, Red Claw Scorpions, Red Scorpions, Sand Scorpions, Tailless Whip Scorpions and Whip Scorpions. Many of these venoms have natural chlorotoxin that have shown indications as effective treatments for disease states such as cancer, Alzheimer's, epilepsy, inflammation and pain management. Scorpiones (Scorpions), Current records show that there are 1,004 species known which presents the researcher a wealth of chlorotoxins with which to analyze for effective treatments to disease states such as cancer, Alzheimer's, epilepsy, inflammation and pain management. The non-disulfide-bridged scorpion venom peptide has become a crucial substance to initiate pharmacologic activities including antibacterial, antifungal, antimalarial, antiviral and anticancer activities.

(8) Specifically, for this example the Blue Scorpion Venom Chlorotoxin venom is extracted by mild Electro-stimulation method, that includes stimulating and scorpion restraining devices. To obtain the venom, the scorpion is subjected to an electrical current from a simple 12-volt battery or low voltage power supply such that the venoms are collected by the electrical stimulation method. Specifically, a series of regular currents are applied to shock the scorpion until the venom is ejected. For that purpose, controlling the scorpion is important, the scorpion is held with forceps by the tail and is then given a shock with electrode connected to the low voltage power source such as a battery or power supply. The venom droplet is recovered in a 1 ml vial after which the extracted venom kept frozen until used.

(9) Then once the venom is extracted and frozen, the venom can be recovered from the freezing process using distilled water and centrifuged. The supernatant can then be lyophilized (freeze-dried), and maintained at 20 C. until used or the lyophilized powder can be added to the phytonutrients powder in the ratio of 1-part venom 10,000 parts by weight phytonutrients extract and mixing thoroughly. The extract from the residue is filtered and concentrated under vacuum. The extract from the residue is vacuum tray dried at 70-80 C. for 12 hours. The extract from the residue is scraped and the dried lumps of the extract from the residue are milled. The natural chlorotoxin is added to the extract/residue and blended homogenously. The extract/residue is sieved and packed. The percentage yield is calculated according to the weight of the raw materials.

(10) The finished powder is made into a nutraceutical supplement in the form of pills that are taken by the patient. However, the final nutraceutical supplement could be formulated into a number of suitable delivery means such as, injectable, oral tablet, oral liquid, topical cream, nasal spray, dry powder aerosols, powder structures for water-soluble drugs, and powder containing time release capsules, since dry powder aerosols is the best route reported for chlorotoxin to cross the blood brain barrier and reach its site in cases of glioma this may be a preferred formulation. The pills can also contain any buffering or inorganic compounds suitable for the pill formation and that does not adversely affect the finished powder of phytonutrients extract and chlorotoxin powder. Typical excipients can be found in Table 1. All pharmaceutical formulations of the venom will be taken as a base in a suitable physiological vehicle to which the processed phytonutrient ingredients will be added and formulated with other inert excipients, stabilizers, and preservatives, as needed. The oral liquid and injectable formulations can be formed by taking the powder of phytonutrients extract and chlorotoxin powder and mixing it with a liquid, wherein the liquid is selected from the group consisting of deionized water, saline, plasma, and interstitial fluid, as well as stabilizers and preservatives, as needed.

(11) The injectable formulations commonly use chemicals used in the production of vaccines. These include suspending fluid (sterile water, saline, or fluids containing protein); preservatives for example (Benzoic acid, salts, Sorbic acid, Parabens and paraben substitutes such as natural products such as grape fruit seed, Benzyl Alcohol, Phenoxyethanol, Sodium Benzoate, Potassium Sorbate) and stabilizers (for example, albumin, phenols, and glycine); and adjuvants or enhancers that help improve the injectable effectiveness. The adjuvants can be selected from the group consisting of aluminum phosphate, aluminum hydroxide, Docosahexaenoic acid, and MF59, an immunologic adjuvant that uses squalene. The oral formulations utilize stabilizers selected from polyols, albumins and other proteins, surfactants, poloxamers, cyclodextrins and preservatives selected from Benzoic acid, salts, Sorbic acid, Parabens and paraben substitutes such as natural products such as grape fruit seed, Benzyl Alcohol, Phenoxyethanol, Sodium Benzoate, Potassium Sorbate. The oral formulations utilize preservatives selected from the group Methyl, ethyl, propyl parabens sodium benzoate, benzoic acid, Methyl paraben/sodium benzoate, Benzyl Alcohol, Phenoxyethanol Sodium Benzoate, Potassium Sorbate and suitable combinations.

(12) The pharmaceutical formulations of the present invention provide a formulation/composition comprising phytonutrients and natural chlorotoxins for targeting cancer in humans, felines and canines, and other pet animals, without any side effects.

(13) Table 1 below illustrates the composition of the formulation/composition comprising phytonutrients and natural chlorotoxins for targeting cancer, Alzheimer's, epilepsy and pain management and excipients that can be utilized in the instant invention:

(14) TABLE-US-00001 TABLE 1 Sl Name of Main function/ No Phytonutrient Derived from % References Pharmaceuticals actives 1 Glycyrrhizic Glycyrrizaglabra 13% Anti-oxidant1 acid 2 Piperine Piper longum 0.54% Bioavailability enhancer, 2 3 Picroside & Picrorhhizakurroa 15% Hepatoprotective 3 kuti koside 4 Tannins Phyllanthusamarus 55% Anti-oxidant4 5 Terminalia 78% chebula 6 36 amino acid Blue scorpion 0.01% Anti-cancerous 5 peptide saliva Excipients 7 Aqua Sucralose Diluting agent and citric acid. 8 Sorbitol 9 Glycerine 10 Sodium Preservatives benzoate 11 Na CMC 12 Honey Sweetener 13 Sucralose Sweetener 14 Citric acid Excipient 15 Water Diluting agent

(15) According to one embodiment of the present invention, Bauhinia variegata bark extract is extracted in hydro-alcohol and used in the anti-cancer composition/formulation. Traditional literature states that, the Bauhinia variegata bark is used in worm infestation, scrofula, cervical lymphadenitis, and wounds. The Bauhinia variegata bark has antimicrobial and wound healing properties. In the phytoceutical approach, hydro-alcohol extract comprises quercitroside, Kaempferol-3-glucoside, lupeol and beta-sitosterolisoquercitroside, rutosidemyricetol glycoside, and Kaempferol glycosides. Stem extracts of Bauhinia variegata comprise phenanthraquinone, 2,7-dimethoxy-3-methyl-9,10 dihydrophenanthrene-1-4, dione (Bauhinione). The extracts from Bauhinia variegate find application in various inflammatory conditions, diabetes, chronic hyperglycemia, antitumor, and cytotoxic activity.

(16) According to one embodiment of the present invention, the main compounds present in the anti-cancer formulation or composition (RX002CxN) are the phyto-complex with natural chlorotoxins. The phyto-complex comprises the extracts of Glycyrrizaglabra, Piper longum, Picrorhiza kurroa, Phyllanthus amarus, Bauhinia variegate, and Terminalia chebula, in predetermined amounts. The chlorotoxins used in the composition are derived from the polypeptide of the blue scorpion saliva. This phyto-complex with natural chlorotoxins act as a potent anti-cancer composition and is active against multi-focal oncological usages with special attention of cancers of blood, bone, colon, cervix, bladder, ovary, breast, prostate, and uterus.

(17) According to one embodiment of the present invention, the following are methods of in vitro assay for anti-cancerous activity of the formulation/composition comprising phytonutrients and natural chlorotoxins for targeting cancer:

Example-1

Sulforhodomine B (SRB) Assay

(18) The experimental procedure for SRB Assay is adopted for analyzing the anti-cancerous activity of the formulation/composition (RX002CxN). The dose response parameters are calculated for each test article. The growth inhibition of 50% (GI 50) is calculated from (TiTz)/(CTz)100=50, which is the drug concentration resulting in a 50% reduction in the drug incubation. The drug concentration resulting in total growth inhibition (TGI) is calculated from Ti=Tz. The Lc50 indicating net loss of cells following treatment is calculated from [(TiTz)/Tz]100=50. The values are calculated for each of these three (3) parameters if the level of activity was reached, however, if the effect is not reached or is exceeded, the values for that parameter are expressed as greater or less than the maximum or minimum concentration tested.

(19) Following are the steps in the SRB assay protocol: a. Preparation of the drug solutions: Confirming the test substance and number with Form A whenever a new drug received. b. Test substance is stored based on the criteria and checked. c. If the molecular weight of test substance is not known, prepare the working solution of the drug (10, 20, 40, 80 microgram/ml) with a specified vehicle. d. If the molecular weight of the test substance is known, then prepare the working solution of the drug 10.sup.7, 10.sup.6, 10.sup.5, 10.sup.4 M concentration with the specified vehicle. e. The working stock of the drug is prepared in appropriate solvent and further dilutions are made as per Table 1 below:

(20) TABLE-US-00002 Drug Concentration L of the L of the L Volume to well working MiLLI-Q Total volume be Added to Molar stock water L the well g/ml concentration A B A B A B A B 10 10.sup.7 10 1 90 999 100 1000 10 10 20 10.sup.6 20 10 80 990 100 1000 10 10 40 10.sup.5 40 100 60 900 100 1000 10 10 80 10.sup.4 80 1000 20 00 100 1000 10 10 f. Prepare the same concentrations of the test substance (the formulation or composition) for the vehicle control and also for the positive control. g. Prepare for the culture plate preparation and drug or composition addition with single cell suspension of the required cell line grown in tissue, followed by counting of cells. The cell count is adjusted according to titration readings, each test substance (composition) is tested in 96 well plates with its four (4) dilutions in triplicate. The culture plates are incubated at 37 C. in carbon dioxide incubator for 24 hours. The drugs with dilutions 10, 20, 40, 80 L are made in triplicate and accordingly culture plates are labeled as: VC: Vehicle Control PC: Positive Control h. The culture plates are labeled and incubated for 48 hours. Experiment is terminated by gently layering the cells in the wells with 50 ml of chilled 30% TCAin case of adherent cells and 50% TCAin case the suspension cell lines for cell fixation. The culture plates are refrigerated at 4 C. for an hour. The culture pates are washed thoroughly with tap water at least five (5) times and air dried. i. The culture plates are stained with 50 L of 0.4% SRB for 20 minutes. The culture plates are washed with 1% acetic acid at least five (5) times and air dried. The bound SRB is eluted with 100 L of Tris buffer for 10 minutes on a shaker. The absorbance is read in the micro titer reader at 540 nm with reference of 690 nm. The optical density of drug treated cells is compared with that of control cells and the growth inhibition is calculated at % values. j. The anti-cancer potential of the formulation/composition (RX002CxN) comprising phytonutrients from phytonutrients bearing plants and natural chlorotoxins for targeting cancer is established through in vivo test or PK/PD study design. The absorption of formulation/composition RX002CxN starts in the duodenum, the first section of the small intestine in higher vertebrates. The peptides, proteins, and minerals present in the formulation/composition RX002CxN are absorbed completely by the human body, within 30 minutes. The main compound in the formulation/composition RX002CxN is a peptide comprising 36 amino acids along with flavonoids. The peptide comprising 36 amino acids pass the blood brain barrier and inhibits glioma cell invasion. The 36 amino acid peptide in the formulation/composition RX002CxN has a cumulative effect, within 24 hours.

Example-2

Activity Criteria

(21) The inhibitory concentration (IC50) for synthetic compounds/formulations is typically 10 g/ml or 10.sup.5M. The inhibitory concentration (IC50) for natural products or plant extracts typically is IC5020 mg/ml, and at any given point concentration, % growth inhibition 50 indicates activity.

(22) Table 2 below illustrates the cell lines and the tissue of origin from which the cell lines are derived:

(23) TABLE-US-00003 Sr Nos Name of the cell line Tissues of origin 1 Colo 205 Colon 2 Hop62 Lung 3 HT29 Colon 4 SiHa Cervix 5 MIAPACA 2 Pancreas 6 DWD Oral 7 T24 Bladder 8 PC3 Prostrate 9 A549 Lung 10 ZR 75 1 Breast 11 A2780 Ovary 12 DU145 Prostate 13 MCF7 Breast 14 K562 Leukemia

(24) According to one embodiment of the present invention, the drug development stages (formulation/composition) are carried out in the following stages:

(25) Drug Discovery: Drug discovery begins with target identification-choosing a biochemical mechanism involved in a disease condition. The drug/composition/formulation candidates, discovered in academic and pharmaceutical/biotech research labs, are tested for their interaction with the drug target. Up to 5,000 to 10,000 molecules for each potential drug candidate are subjected to a rigorous screening process that includes functional genomics and/or proteomics, as well as other screening methods. After the researchers/scientists confirm interaction of the drug/composition with a target, the target is validated by checking for drug/composition activity versus the disease condition for which the drug/composition is being developed. After careful review, one or more lead compounds are chosen.

(26) Ideation: This stage is primitive and creates and idea for how to lead with drug trials.

(27) Target identification: This stage deals with how to fix the target, molecules, and precursors.

(28) Product Characterization: When the candidate molecule shows promise as a therapeutic, it must be characterized; the molecule's size, shape, strengths and weaknesses, preferred conditions for maintaining function, toxicity, bioactivity, and bioavailability must be determined. Characterization studies will undergo analytical method development and validation. Early stage pharmacology studies help to characterize the underlying mechanism of action of the compound, including phytoceuticals.

(29) Formulation, Delivery, Packaging Development: Drug developers must devise a formulation that ensures the proper drug delivery parameters. It is critical to begin looking ahead to clinical trials at this phase of the drug development process. Drug formulation and delivery may be refined continuously until, and even after, the drug's final approval. Scientists determine the drug's stability in the formulation itself, and for all the parameters involved with storage and shipment, such as heat, light, and time. The formulation must remain potent and sterile; and it must also remain safe (nontoxic). It may also be necessary to perform leachable on containers or packaging.

(30) Pharmacokinetics and Drug Disposition: Pharmacokinetic (PK) and ADME (Absorption/Distribution/Metabolism/Excretion) studies provide useful feedback for formulation scientists. PK studies yield parameters such as AUC (area under the curve), Cmax (maximum concentration of the drug in blood), and Tmax (time at which Cmax is reached). Later on, this data from animal PK studies is compared to data from early stage clinical trials to check the predictive power of animal models.

(31) Preclinical Toxicology Testing and IND Application: Preclinical testing analyzes the bioactivity, safety, and efficacy of the formulated drug product. This testing is critical to a drug's eventual success and as such, is scrutinized by many regulatory entities. During the preclinical stage of the development process, plans for clinical trials and an Investigational New Drug (IND) application are prepared. Studies taking place during the preclinical stage should be designed to support the clinical studies that will follow.

(32) The main stages of preclinical toxicology testing are:

(33) Acute StudiesAcute toxicology (tox) studies analyze the effects of one or more doses administered over a period of up to 24 hours. The aim of acute tox studies is to determine toxic dose levels and observe clinical indications of toxicity. Usually, at least two mammalian species are tested. Data from acute tox studies help determine doses for repeated dose studies in animals and Phase I clinical trials in humans.

(34) Repeated Dose StudiesDepending on the duration of the studies, repeated dose studies may be referred to as subacute, subchronic, or chronic. The specific duration should anticipate the length of the clinical trial that will be conducted on the new drug. Again, two species are typically required.

(35) Genetic Toxicity StudiesThese studies assess the likelihood that a drug compound is mutagenic or carcinogenic. Procedures such as the Ames Test (conducted in bacteria) detect genetic changes. DNA damage is assessed in tests using mammalian cells such as the Mouse Micronucleus Test. The Chromosomal Aberration Test and similar procedures detect damage at the chromosomal level.

(36) Reproductive Toxicity StudiesSegment I reproductive fox studies analyze the effects of the drug on fertility. Segment II and III studies detect effects on embryonic and post-natal development. In general, reproductive tox studies must be completed before a drug can be administered to women of child-bearing age.

(37) Carcinogenicity StudiesCarcinogenicity studies are usually needed only for drugs intended for chronic or recurring conditions. They are time consuming and expensive, and must be planned for early in the preclinical testing process.

(38) Toxicokinetic StudiesThese are typically similar in design to PK/ADME studies except that they use much higher dose levels. They examine the effects of toxic doses of the drug and help estimate the clinical margin of safety. There are numerous FDA and ICH guidelines that give a wealth of detail on the different types of preclinical toxicology studies and the appropriate timing for them relative to IND and NDA or BLA filings.

(39) Bioanalytical Testing: Bioanalytical laboratory work and bioanalytical method development supports most of the other activities in the drug development process. The bioanalytical work is key to proper characterization of the molecule, assay development, developing optimal methods for cell culture or fermentation, determining process yields, and providing quality assurance and quality control for the entire development process. It is also critical for supporting preclinical toxicology/pharmacology testing and clinical trials.

(40) Clinical Trials: Clinical studies are grouped according to their objective into three types or phases:

(41) Phase I Clinical Development (Human Pharmacology)Phase I studies are used to evaluate pharmacokinetic parameters, tolerance and safety, generally in healthy volunteers. These studies include initial single-dose studies, dose escalation and short-term repeated-dose studies.

(42) Phase II Clinical Development (Therapeutic Exploratory)Phase II clinical studies are small-scale trials to evaluate a drug's preliminary efficacy and side effect profile. Additional safety and clinical pharmacology studies are also included in this category.

(43) Phase III Clinical Development (Therapeutic Confirmatory)Phase III studies are large-scale clinical trials for safety and efficacy in large patient populations. While phase III studies are in progress, preparations are made for submitting the Biologics License Application (BLA), filing under the FDA Nutraceutical Act or the New Drug Application (NDA).

(44) The foregoing description of the specific embodiments will so fully reveal the general nature of the embodiments herein that others can, by applying current knowledge, readily modifying and/or adapting for various applications, 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.

(45) It is to be understood that the phraseology or terminology employed herein is 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 appended claims.

(46) Although the embodiments herein are described with various specific embodiments, it will be obvious for a person skilled in the art to practice the embodiments herein with modifications. The scope of the embodiments will be ascertained by the claims to be submitted at the time of filing a complete specification.

Advantages of the Invention

(47) The embodiments of the present invention provide a formulation comprising phytonutrients from phytonutrient-bearing plants and natural chlorotoxins for targeting cancer, infection, inflammation, and pain, without any side effects.

(48) The embodiments of the present invention provide a method for the synthesis of a formulation comprising phytonutrients from phytonutrient-bearing plants and natural chlorotoxins for targeting cancer, infection, inflammation, and pain.

(49) The embodiments of the present invention provide a formulation comprising phytonutrients and natural chlorotoxins for minimizing tumor growth.

(50) The embodiments of the present invention provide a formulation comprising phytonutrients from phytonutrient-bearing plants and natural chlorotoxins for minimizing secondary metastasis by apoptosis of variety of cancer cells by a pattern of DNA fragmentation.

(51) The embodiments of the present invention provide a formulation comprising phytonutrients from phytonutrients bearing plants and natural chlorotoxins for arresting cancer cell growth.

(52) The embodiments of the present invention provide a formulation comprising phytonutrients from phytonutrient-bearing plants and natural chlorotoxins for prevention of angiogenesis in cancer cells.

(53) The embodiments of the present invention provide a formulation comprising phytonutrients from phytonutrient-bearing plants and natural chlorotoxins for regulating the process of phosphorylation that affects the p53 tumor suppressor protein leading to cell cycle arrest and apoptotic cell death.

(54) The embodiments of the present invention provide a formulation comprising phytonutrients from phytonutrient-bearing plants and natural chlorotoxins for arresting the growth of cancer cell in the G1 phase of the cell cycle, changing in mitogenic signals specific to malignant cells, and providing a universal effect on different forms of cancer.

(55) The pharmaceutical formulations of the present invention provide a formulation/composition comprising phytonutrients and natural chlorotoxins for targeting cancer, infection, inflammation and pain in humans, felines and canines, and other pet animals without any side effects.

(56) The foregoing description of the specific embodiments will so fully reveal the general nature of the embodiments herein that others can, by applying current knowledge, readily modify and/or adapt for various applications 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.

(57) 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 appended claims.

(58) Although the embodiments herein are described with various specific embodiments, it will be obvious for a person skilled in the art to practice the embodiments herein with modifications. The scope of the embodiments will be ascertained by the claims to be submitted at the time of filing a complete specification.

(59) It should be understood that the foregoing description is only illustrative of the invention. Various alternatives and modifications can be devised by those skilled in the art having the benefit of this disclosure, without departing from the invention. Accordingly, the invention is intended to embrace all such alternatives, modifications and variances.

(60) Certain exemplary embodiments of the disclosure may be described. Of course, the embodiments may be modified in form and content, and are not exhaustive, i.e., additional aspects of the disclosure, as well as additional embodiments, will be understood and may be set forth in view of the description herein. Further, while the invention may be susceptible to various modifications and alternative forms, specific embodiments have been shown by way of example in the drawings and will be described in detail herein. However, it should be understood that the invention is not intended to be limited to the particular forms disclosed. Rather, the invention is to cover all modifications, equivalents and alternatives falling within the spirit and scope of the invention.

(61) To the extent that the term includes or including is used in the specification or the claims, it is intended to be inclusive in a manner similar to the term comprising as that term is interpreted when employed as a transitional word in a claim. Furthermore, to the extent that the term or is employed (e.g., A or B) it is intended to mean A or B or both. When the applicants intend to indicate only A or B but, not both then the term only A or B but not both will be employed. Thus, use of the term or herein is the inclusive, and not the exclusive use. See, Bryan A. Garner, A Dictionary of Modern Legal Usage 624 (2d, Ed. 1995). Also, to the extent that the terms in or into are used in the specification or the claims, it is intended to additionally mean on or onto. Furthermore to the extent the term connect is used in the specification or claims, it is intended to mean not only directly connected to, but also indirectly connected to such as connected through another component or components.

(62) While the present application has been illustrated by the description of embodiments thereof, and while the embodiments have been described in considerable detail, it is not the intention of the applicants to restrict or in any way limit the scope of the appended claims to such detail. Additional advantages and modifications will readily appear to those skilled in the art. Therefore, the application, in its broader aspects, is not limited to the specific details, the representative apparatus and method, and illustrative examples shown and described. Accordingly, departures may be made from such details without departing from the spirit or scope of the applicant's general inventive concept.