Description:FORM 2
THE PATENTS ACT, 1970
(39 of 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10; rule 13)

TITLE OF INVENTION:
OIL-BASED COMPOSITION EXHIBITING CYTOTOXIC ACTIVITY AGAINST MALIGNANT CELLS AND METHOD OF PREPARATION THEREOF
APPLICANTS:
LIFEIS SPEED PRIVATE LIMITED
An Indian company having address as:
Unit no. 546, 5th floor, IJMIMA Complex, Off Link, Road, Malad West,
Mumbai 400064, Maharashtra, India

PREAMBLE TO THE DESCRIPTION
The following complete specification particularly describes the disclosure and the manner in which it is to be performed. 
TECHNICAL FIELD
The present disclosure relates to storage management system. More particularly, the present disclosure adjustable shelf device, system and method for automated storage of products.
BACKGROUND
The global demand for nutraceuticals dietary supplements that offer both nutritional and therapeutic value has grown substantially in recent years. This rise is driven by increased consumer awareness of preventive health, aging populations, and lifestyle-related disorders such as cancer, cardiovascular diseases, and neurodegenerative conditions. As a result, naturally derived bioactive ingredients have gained significant attention for their potential in managing oxidative stress, supporting immune health, and promoting general wellness.
Among such ingredients, vegetable oils rich in bioactive compounds have been widely researched. Wheat germ oil (WGO) and rice bran oil (RBO) are two such natural oils recognized for their functional and nutritional properties. Each is derived from by-products of staple cereal crops and has been traditionally used in both food and cosmetic industries. Their high content of essential fatty acids, antioxidants, vitamins, and phytosterols positions them as valuable constituents in nutraceutical and functional food formulations.
Wheat germ oil is extracted from the germ of the wheat kernel and is notably rich in natural vitamin E (particularly alpha-tocopherol), octacosanol, and omega-3 and omega-6 fatty acids. It also contains a variety of bioactive components such as policosanols and phytosterols, which have been investigated for their antioxidant, cardioprotective, anti-inflammatory, and anti-aging effects. The antioxidant properties of WGO make it a candidate for applications involving the mitigation of oxidative cellular damage.
Rice bran oil, obtained from the outer husk (bran) of rice grains, contains a unique profile of tocopherols, tocotrienols, γ-oryzanol, and unsaturated fatty acids. These compounds have been shown to exhibit cholesterol-lowering effects, antioxidant activity, and potential anti-cancer properties. Additionally, γ-oryzanol has been studied for its ability to regulate lipid metabolism and suppress oxidative stress-induced damage in tissues.
Despite the individual health benefits of WGO and RBO, current nutraceutical formulations tend to utilize these oils independently. There is limited exploration in the prior art regarding their combined use in fixed-dose ratios designed to exploit potential synergistic interactions between their respective bioactive components. While the literature suggests that individual phytochemicals present in WGO or RBO may have therapeutic relevance, the effect of blending these oils in specific proportions to optimize bioavailability, stability, and biological activity has not been adequately investigated or commercialized.
Moreover, combining lipophilic compounds from different sources often presents formulation challenges, including phase separation, oxidation, and loss of activity. Therefore, the development of a stable, homogeneous, and bioactive oil-based blend remains an area of active research in the nutraceutical and pharmaceutical sectors.
In particular, the emerging interest in natural alternatives to support conventional therapies in chronic and degenerative diseases such as cancer has prompted deeper examination of natural oil combinations for their cytoprotective or cytotoxic effects, depending on the application. In this context, formulations that demonstrate selective activity i.e., cytotoxic to cancerous cells while sparing healthy cells are of significant scientific and commercial interest.
Therefore, there exists a need for a defined, fixed-ratio combination of wheat germ oil and rice bran oil, formulated in a pharmaceutically or nutraceutically acceptable dosage form, that offers enhanced functional performance, reduced toxicity, and broad-spectrum biological support, particularly in the context of antioxidant protection, cellular health, and potentially adjunctive therapy in chronic diseases.
SUMMARY
In one aspect of the present disclosure, an oil-based composition is provided.
The oil-based composition includes wheat germ oil in an amount of about 40-50%, said oil includes naturally occurring tocopherols, linoleic acid, oleic acid, palmitic acid, phytosterols, and essential fatty acids.
The oil-based composition further includes rice bran oil in an amount of about 25-35%, said oil includes γ-oryzanol, tocotrienols, phytosterols, and unsaturated fatty acids including oleic and linoleic acids.
In some aspects of the present disclosure, the oil-based composition further includes glycerin in an range of about 10-15% of the oil-based composition.
In some aspects of the present disclosure, the oil-based composition further includes Sorbitol in an amount of about 5-10% of the oil-based composition,
In some aspects of the present disclosure, the oil-based composition further includes purified water in an amount of about 2-5% of the oil-based composition.
In some aspects of the present disclosure, the wheat germ oil and rice bran oil are mixed by pipetting and vortexing until uniform consistency is achieved.
In some aspects of the present disclosure, the oils are sterilized by filtration prior to in vitro application.
In second aspect of the present disclosure, a method for preparing a homogeneous oil-based composition is provided.
The method includes measuring Wheat Germ Oil in an amount of about 300 mg. The method further includes measuring Rice Bran Oil in an amount of about 200 mg. The method further includes combining the Wheat Germ Oil and Rice Bran Oil using pipetting. The method further includes vortexing the combined mixture for a predetermined time at room temperature and atmospheric pressure to form a homogeneous oil blend.
In some aspects of the present disclosure, the vortex mixing is at a speed ranging from 2500 to 3000 RPM for a period of 30 to 60 seconds to form a stable and homogeneous oil blend.
In some aspects of the present disclosure, the oil-based composition are filtered through a 0.22 µm membrane prior to filling.
In some aspects of the present disclosure, the oil-based composition are sterilized by filtration prior to in vitro application.
BRIEF DESCRIPTION OF DRAWINGS
The accompanying drawings, which are incorporated in and constitute a part of this specification, show certain aspects of the subject matter disclosed herein and, together with the description, help explain some of the principles associated with the disclosed implementations. In the drawing,
Figure 1 illustrates a flowchart that depicts a method for preparing oil-based composition, in accordance with an aspect of the present disclosure;
Figure 2A and 2B illustrates a graph representing results associated with microtiter plate assay for determination of minimum inhibitory concentration of gram positive and gram-negative bacteria, in accordance with an aspect of the present disclosure;
Figure 3 illustrates a graph representing results associated with cytotoxicity analysis, in accordance with an aspect of the present disclosure;
Figure 4 illustrates a graph representing results associated with antioxidative test, in accordance with an aspect of the present disclosure; and
Figure 5 illustrates a graph representing results associated with GBM cell assay, in accordance with an aspect of the present disclosure.
To facilitate understanding, like reference numerals have been used, where possible, to designate like elements common to the figures.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
Various embodiments of the disclosure are discussed in detail below. While specific implementations are discussed, it should be understood that this is done for illustration purposes only. A person skilled in the relevant art will recognize that other components and configurations may be used without parting from the spirit and scope of the disclosure. Thus, the following description and drawings are illustrative and are not to be construed as limiting. Numerous specific details are described to provide a thorough understanding of the disclosure. However, in certain instances, known details are not described in order to avoid obscuring the description.
References to one or an embodiment in the present disclosure can be references to the same embodiment or any embodiment; and such references mean at least one of the embodiments.
Reference to "one embodiment", "an embodiment", “one aspect”, “some aspects”, “an aspect” means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the disclosure. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments. Moreover, various features are described which may be exhibited by some embodiments and not by others.
The terms used in this specification generally have their ordinary meanings in the art, within the context of the disclosure, and in the specific context where each term is used. Alternative language and synonyms may be used for any one or more of the terms discussed herein, and no special significance should be placed upon whether or not a term is elaborated or discussed herein. In some cases, synonyms for certain terms are provided.
Additional features and advantages of the disclosure will be set forth in the description which follows, and in part will be obvious from the description, or can be learned by practice of the herein disclosed principles. The features and advantages of the disclosure can be realized and obtained by means of the instruments and combinations particularly pointed out in the appended claims. These and other features of the disclosure will become more fully apparent from the following description and appended claims or can be learned by the practice of the principles set forth herein.
As mentioned before, there exists a need for a defined, fixed-ratio combination of wheat germ oil and rice bran oil, formulated in a pharmaceutically or nutraceutically acceptable dosage form, that offers enhanced functional performance, reduced toxicity, and broad-spectrum biological support, particularly in the context of antioxidant protection, cellular health, and potentially adjunctive therapy in chronic diseases. The present disclosure, therefore, provides a nutraceutical composition comprising a fixed-dose combination of wheat germ oil (WGO) and rice bran oil (RBO), respectively. The composition is formulated as a homogeneous blend, suitable for oral administration, and intended to deliver synergistic health benefits derived from the complementary bioactive profiles of WGO and RBO.
In one aspect of the present disclosure, an oil-based composition includes wheat germ oil and rice bran oil.
The wheat germ oil may be in a range of about 40-50%. In some aspects of the present disclosure, the wheat germ oil may include naturally occurring tocopherols, linoleic acid, oleic acid, palmitic acid, phytosterols, and essential fatty acids.
In some aspects of the present disclosure, the wheat germ oil may be present in the oil-based composition of about 45.11%.
In some aspects of the present disclosure, the rice bran oil in an amount of about 25-35%. In some aspects of the present disclosure, the oil includes γ-oryzanol, tocotrienols, phytosterols, and unsaturated fatty acids including oleic and linoleic acids.
In some aspects of the present disclosure, the rice bran oil may be present in the oil-based composition of about 30.08%.
The oil-based composition may further include glycerin, sorbitol and purified water.
In some aspects of the present disclosure, the glycerin may be in an range of about 10-15% of the oil-based composition. In some aspects of the present disclosure, the Glycerin may be present in the oil-based composition of about 13.53%.
In some aspects of the present disclosure, the sorbitol may be in an amount of about 5-10% of the oil-based composition. In some aspects of the present disclosure, the sorbitol may be present in the oil-based composition of about 7.52%.
In some aspects of the present disclosure, the purified water may be in an amount of about 2-5% of the oil-based composition. In some aspects of the present disclosure, the purified water may be present in the oil-based composition of about 3.76%.
In an exemplary scenario, when a room of the oil-based composition may be 100 percent which is 655mg. The Wheat Germ Oil may be in an amount of about 300 mg. In some aspects of the present disclosure, the Wheat Germ Oil may include naturally occurring tocopherols, linoleic acid, oleic acid, palmitic acid, phytosterols, and essential fatty acids. The Rice Bran Oil may be in an amount of about 200 mg. In some aspects of the present disclosure, rice bran oil may include γ-oryzanol, tocotrienols, phytosterols, and unsaturated fatty acids including oleic and linoleic acids. In some aspects of the present disclosure, the glycerin may be in an about 80 mg. In some aspects of the present disclosure, the sorbitol may be in an about 46.88mg. In some aspects of the present disclosure, the purified water may be in an amount of about 20mg.
In some aspects of the present disclosure, the wheat germ oil and rice bran oil may be mixed by pipetting and vortexing until uniform consistency is achieved.
In some aspects of the present disclosure, the oils may be sterilized by filtration prior to in vitro application.
In some aspects of the present disclosure, the combination may demonstrate lower cytotoxicity on Vero cell lines relative to the individual oils when tested at equivalent concentrations.
In some aspects of the present disclosure, the combination may exhibits % viability of Vero cells exceeding 80% at the said concentrations.
In some aspects of the present disclosure, the synergistic antioxidative efficacy may be compared against a standard positive control comprising ascorbic acid.
Figure 1 illustrates a method 100 for preparing a homogeneous oil-based composition, in accordance with an aspect of the present disclosure. The method 100 may include the following steps:
At step 102, the method 100 may include measuring Wheat Germ Oil in an amount of about 300 mg.
At step 104, the method 100 may include measuring Rice Bran Oil in an amount of about 200 mg.
At step 106, the method 100 may include combining the Wheat Germ Oil and Rice Bran Oil using pipetting.
At step 108, the method 100 may include vortexing the combined mixture for a predetermined time at room temperature and atmospheric pressure to form a homogeneous oil blend.
At step 110, the method 100 may include adding the sorbitol, the glycerin, and the purified water into the obtained homogeneous oil blend.
In some aspects of the present disclosure, the vortex mixing is at a speed ranging from 2500 to 3000 RPM for a period of 30 to 60 seconds to form a stable and homogeneous oil blend.
In some aspects of the present disclosure, the oil-based composition are filtered through a 0.22 µm membrane prior to filling. In some aspects of the present disclosure, the oil-based composition are sterilized by filtration prior to in vitro application.
Example 1: method for preparing a homogeneous oil-based composition
300mg of rice bran oil was measure and added to a container having 200mg of rice bran oil. The mixture was combines by way of pipetting for 5 minutes at room temperature and atmospheric pressure, and the homogeneous oil blend was obtained. 80mg of glycerin, 46.88mg of sorbitol, and 20mg of purified water was added into the obtained homogeneous oil blend.
Example 2: Microtiter plate assay for determination of minimum inhibitory concentration
The different concentrations of curcumin and combinations of wheat germ oil (WGO) and rice bran oil (RBO) were incubated with 1×10⁶ CFU/mL of the indicator organism in each well of a 96-well microtiter plate. Bacterial cultures (gram positive and gram negative) without any antimicrobial agents and media alone were used as negative and baseline controls, respectively. The plates were incubated at 37°C for 18 hours, after which the wells were examined for bacterial growth inhibition in comparison to the untreated bacterial growth control and shown in figure 2A and 2B. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of the tested formulation at which more than 90% inhibition of bacterial growth was observed. The assay was performed in triplicates and repeated three times to ensure reproducibility and reliability of the results. The % inhibition was calculated by following formula:
% 𝑖𝑛ℎ𝑖𝑏𝑖𝑡𝑖𝑜𝑛 = [1 − {𝑂𝐷𝑜𝑓𝑡𝑟𝑒𝑎𝑡𝑒𝑑𝑐𝑒𝑙𝑙𝑠/𝑂𝐷𝑜𝑓𝑢𝑛𝑡𝑟𝑒𝑎𝑡𝑒𝑑𝑐𝑒𝑙𝑙𝑠}]100
Example 3: 3-(4, 5-dimethylthiazol-2- yl)-2, 5-diphenyltetrazolium bromide (MTT) assay for cytotoxicity analysis
The cytotoxicity of the individual oils, as well as their combinations, was evaluated using the MTT assay on Vero epithelial cells derived from the African green monkey. The cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and incubated at 37°C with 5% CO₂ to maintain optimal growth conditions. A cell suspension with a concentration of 1×10⁵ CFU/mL was seeded into 96-well plates and incubated for 24 hours to allow for cell adherence. The test compounds, which were filter-sterilized, were added to the adhered cells and incubated for another 24 hours under identical conditions. Following this, MTT reagent (5 mg/mL) was introduced to each well, and the plates were incubated for an additional 4 hours at 37°C to allow the formation of formazan crystals by metabolically active cells. The resulting absorbance was measured at 570 nm using a 96-well plate reader (Bio-Rad, USA) and shown in figure 3. The cytotoxic effect of the compounds was calculated by comparing the absorbance values of treated wells (AbsT) to those of the untreated control wells (AbsU) using the formula:
% Viability = (AbsT / AbsU) × 100
The results revealed that all formulations, where the combinations, exhibited high cell viability, indicating low cytotoxicity. In most cases, cell viability exceeded 85%, suggesting that the tested oil-based compositions were biocompatible and suitable for further biological or therapeutic applications. All experiments were conducted in triplicates and repeated three times to ensure reproducibility and statistical reliability.
Example 4: Antioxidative test
The antioxidative potential of wheat germ oil (WGO), rice bran oil (RBO), and their combination was assessed using the DPPH free radical scavenging assay. In the assay, 1.0 mL of 0.1 mM DPPH solution was mixed with 1.0 mL of each test sample individually (WGO and RBO) as well as in combination. A mixture of 1.0 mL DPPH and 1.0 mL of ascorbic acid was used as the positive control, serving as a reference for maximum antioxidant activity. All mixtures were incubated in the dark for 30 minutes at room temperature to prevent light-induced degradation of DPPH. The decrease in absorbance, which indicated radical scavenging, was measured at 517 nm using a UV-Vis spectrophotometer and shown in figure 4. The antioxidant activity was quantified using the formula:
% Inhibition = [(A_control - A_sample) / A_control] × 100
The results showed that both WGO and RBO exhibited measurable antioxidant activity, with the combination of WGO + RBO demonstrating higher % inhibition compared to the oils tested individually. However, ascorbic acid (positive control) showed the highest % inhibition, as expected, confirming the sensitivity and validity of the assay. The above result suggested that the combination of WGO and RBO acted synergistically, enhancing free radical scavenging ability. This indicated potential for use of these oils in formulations aimed at oxidative stress reduction or antioxidant-based therapeutic applications.
Example 4: GBM cell assay
The GBM cell assay was conducted using the same experimental procedure as the MTT cytotoxicity assay, with the only variation being the use of glioblastoma multiforme (GBM) cell lines instead of Vero cells. The assay aimed to assess the anti-proliferative effects of wheat germ oil (WGO), rice bran oil (RBO), and their combinations on cancerous brain cells.
After incubation with the test compounds for 24 hours, followed by treatment with MTT reagent and subsequent absorbance measurement at 570 nm, the % viability of GBM cells was calculated. The results demonstrated that WGO and RBO, particularly in combination, induced a concentration-dependent decrease in GBM cell viability. Compared to Vero cells, the oils showed greater cytotoxicity against GBM cells, suggesting selective anticancer potential. In several combinations, the cell viability dropped below 60%, indicating moderate to strong anticancer activity. In parallel, the viscosity profiles of the oils at different temperatures were evaluated to understand their physical characteristics, which may influence formulation and delivery. At 20°C, WGO exhibited a viscosity of approximately 65–72 cP, whereas RBO showed higher viscosity ranging between 90–100 cP. At 40°C, the viscosity of both oils decreased substantially WGO dropped to ~35–40 cP, and RBO to ~55–60 cP and shown in figure 5.
This temperature-dependent reduction in viscosity suggested that heating the oils before formulation could enhance their miscibility and uniform distribution in biological media, improving both homogeneity and bioavailability during cellular exposure. The GBM assay results, supported by the viscosity data, indicated that thermally optimized oil blends of WGO and RBO could be promising candidates for anticancer applications, especially in formulations targeting brain tumors like glioblastoma.
The foregoing discussion of the present disclosure has been presented for purposes of illustration and description. It is not intended to limit the present disclosure to the form or forms disclosed herein. In the foregoing Detailed Description, for example, various features of the present disclosure are grouped in one or more aspects, configurations, or aspects for the purpose of streamlining the disclosure. The features of the aspects, configurations, or aspects may be combined in alternate aspects, configurations, or aspects other than those discussed above. This device of disclosure is not to be interpreted as reflecting an intention the present disclosure requires more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive aspects lie in less than all features of a single foregoing disclosed aspect, configuration, or aspect. Thus, the following claims are hereby incorporated into this Detailed Description, with each claim standing on its own as a separate aspect of the present disclosure.
Moreover, though the description of the present disclosure has included description of one or more aspects, configurations, or aspects and certain variations and modifications, other variations, combinations, and modifications are within the scope of the present disclosure, e.g., as may be within the skill and knowledge of those in the art, after understanding the present disclosure. It is intended to obtain rights which include alternative aspects, configurations, or aspects to the extent permitted, including alternate, interchangeable and/or equivalent structures, functions, ranges, or steps to those claimed, whether or not such alternate, interchangeable and/or equivalent structures, functions, ranges or steps are disclosed herein, and without intending to publicly dedicate any patentable subject matter
, Claims:I/We Claim:
1. An oil-based composition exhibiting cytotoxic activity against malignant cells, comprising:
Wheat Germ Oil in an amount of about 40-50%, said oil comprising naturally occurring tocopherols, linoleic acid, oleic acid, palmitic acid, phytosterols, and essential fatty acids; and
Rice Bran Oil in an amount of about 25-35%, said oil comprising γ-oryzanol, tocotrienols, phytosterols, and unsaturated fatty acids including oleic and linoleic acids.

2. The oil-based composition as claimed in claim 1, further comprising Glycerin in an range of about 10-15% of the oil-based composition

3. The oil-based composition as claimed in claim 1, further comprising Sorbitol in an amount of about 5-10% of the oil-based composition

4. The oil-based composition as claimed in claim 1, further comprising purified water in an amount of about 2-5% of the oil-based composition.

5. The oil-based composition as claimed in claim 1, wherein the wheat germ oil and rice bran oil are mixed by pipetting and vortexing until uniform consistency is achieved.

6. The oil-based composition as claimed in claim 1, wherein the oils are sterilized by filtration prior to in vitro application.

7. A method (100) for preparing a homogeneous oil-based composition, the method (100) comprising:
measuring (102) Wheat Germ Oil in an amount of about 300 mg;
measuring (104) Rice Bran Oil in an amount of about 200 mg;
combining (106) the Wheat Germ Oil and Rice Bran Oil using pipetting; and
vortexing (108) the combined mixture for a predetermined time at room temperature and atmospheric pressure to form a homogeneous oil blend.

8. The method (100) as claimed in claim 7, wherein the vortex mixing is at a speed ranging from 2500 to 3000 RPM for a period of 30 to 60 seconds to form a stable and homogeneous oil blend.

9. The method (100) as claimed in claim 7, wherein the oil-based composition are filtered through a 0.22 µm membrane prior to filling.

10. The method (100) as claimed in claim 7, wherein the oil-based composition are sterilized by filtration prior to in vitro application.

Dated this 26th Day of July 2025

Prasad Prabhakar Karhad
Agent for the Applicant
IN/PA-2352