Priority Claim:

[0001] This application claims priority from the provisional application numbered 202111056096 filed with Indian Patent Office, Chennai on 3 ^rd February 2022 entitled “A phytochemical formulation for the treatment of joint inflammation”, the entirety of which is expressly incorporated herein by reference.

Preamble to the description

[0002] The following specification describes the invention and the manner in which it is to be performed:

Description of the invention

Technical field of the invention

[0003] The present invention relates to a formulation of combination of phytoconstituents for treatment of joint inflammation. More particularly, the invention relates to a formulation of combination of phytoconstituents namely curcumin from Curcuma longa, Boswellia serrata extract, cinnamon oil extract, total gingerols from ginger extract and turmeric extract forming a complex for treatment of joint inflammation.

Background of the invention

[0004] Inflammation is part of the process by which the immune system defends the body from harmful agents such as bacteria, viruses or other microbes. A variety of foreign agents such as pathogens, damaged cells, toxic chemicals, could induce a biological response from the body by triggering the immune system causing inflammation.

[0005] Inflammation is broadly classified into acute and chronic inflammation. Acute inflammation is the initial response of the body to harmful stimuli and is achieved by the increased movement of plasma and leukocytes from the blood into the injured tissues. Chronic inflammation also known as prolonged inflammation leads to a progressive shift in the type of cells present at the site of inflammation, such as mononuclear cells and is characterized by simultaneous destruction and healing of the tissue from the inflammatory process.

[0006] Inflammation is generally characterized by redness, heat, swelling, pain and loss of function due to the release of inflammatory mediators to the site of injury or infection. Several chronic diseases show inflammation during the initial stages including cardiovascular and bowel diseases, diabetes, arthritis, and cancer show inflammation of the tissues during early diagnosis. The mechanism of action of inflammation begins with the identification of the undesirable stimuli by cell surface pattern receptors, activating the inflammatory pathways leading to the release of inflammatory markers and the release of inflammatory cells.

[0007] Arthritis is an autoimmune disorder in which the body attacks its own tissues due to hyperactivity of the immune system. Arthritis is characterized by joint pain due to inflammation in the lining of the joints. In addition, it is also associated with Joint redness, joint swelling, joint warmth, joint tenderness, limping, lack of mobility of the joints, stiffness and weakness.

[0008] The most common types of arthritis are osteoarthritis and rheumatoid arthritis. Osteoarthritis causes break down of cartilage whereas the rheumatoid arthritis is a disease in which the immune system attacks the joints, beginning with the lining of joints. In addition, the infections or underlying disease, such as psoriasis or lupus also cause other types of arthritis.

[0009] The goal of treatment for arthritis is primarily by reducing the pain and inflammation surrounding the joints. Conventionally, Nonsteroidal anti- inflammatory drugs (NSAIDs) such as naproxen, ibuprofen, and aspirin, corticosteroids etc. These drugs may help relieve symptoms of rheumatoid arthritis, psoriasis, and other similar autoimmune reactions. However, they can also leave a person’s body less able to fight an infection if it occurs and long-term use of corticosteroids is harmful and may result in ulcers, nausea, abdominal pain, gastrointestinal bleeding, tinnitus, etc.

[0010] The use of allopathic medicines results in good improvements but in order to reduce the side effects and improve the quality of life, herbal or complementary medicines are also explored for their beneficial effects in patients with arthritis.

[0011] The alternate treatment approaches such as ayurvedic and homeopathic systems of medicine focus on healing the major cause of the ailment than symptomatic treatment and hence results in fewer side effects compared to allopathic medicines.

[0012] The Patent Application WO2019097417A2 entitled “A pharmaceutical composition made from hydrophobic phytochemicals dispersed in sesame oil to enhance bioactivity” discloses a pharmaceutical composition and method to increase the bioactivity and bioavailability of plant compound having hydrophobic phytochemicals which have poor bioavailability due to poor water solubility. The dispersions of extracts of Boswellia and/or Curcuma longa in sesame seed oil. Analgesic and anti-inflammatory composition having a) an extract of Boswellia, b) one or more of an extract of Curcuma longa, a curcuminoid composition and combinations thereof, and c) a sesame seed oil. The analgesic and antiinflammatory composition has particles having a particle size ranging from less than about 20 micrometers to about 1 micrometer. The analgesic and antiinflammatory composition provides a total combined specific surface area provided by particles in the composition ranging from about 900 m ² per kilogram to about 3000 m2 per kilogram of the composition. The invention also discloses a method of preparing the compositions and methods of treating inflammation and musculoskeletal disorders. [0013] The Patent Application W02011125070A2 entitled “Formulation of curcumin with enhanced bioavailability of curcumin and method of preparation and treatment thereof” discloses a formulation of curcuminoid with essential oil of turmeric to enhance the bioavailability and to augment the biological activity of curcumin, wherein curcumin is the main constituent of curcuminoid and Ar- turmerone is the main constituent of the essential oil of turmeric. The use of the formulation in the manufacture of medicament for the treatment of a variety of diseases is also disclosed.

[0014] The Patent Application US2016166516A1 entitled “Novel curcuminoid formulations and related methods of treatment” discloses a novel curcuminoid formulations which have improved aqueous solubility and bioavailability when compared to known curcuminoid dosage forms. The formulations of the invention exhibit an enhanced intestinal absorption of curcuminoids, a slower curcuminoid metabolism and a reduced rate of systemic curcuminoid elimination. Additionally, upon administration in vivo, the formulations described and claimed herein produce a high, sustained level of the curcuminoid metabolite tetrahydrocurcumin. For example, at about eight to about ten hours after oral administration to a human, tetrahydrocurcumin plasma levels of between about 50 ng/mL to about 175 ng/mL are detectable, even when that metabolite is absent from the administered formulation. Novel processes for making curcuminoid formulations which have improved aqueous solubility and oral bioavailability, and methods of treating a wide variety of inflammatory, immune and neurogenerative disorders and cancers, are also provided.

[0015] The Patent Application CN104717969A entitled “Novel formulations and uses for curcuma extracts ” discloses the formulations and uses of extracts of Curcuma longa plants for safe use topically, orally, rectally, or vaginally, for example, are provided. A sunscreen containing an extract of Curcuma longa is provided, the sunscreen having an absorption that spans the UVA and UVB ranges in a manner that meets updated FDA recommendations without requiring the addition of titanium dioxide. A process of producing the extract is also provided, using an extraction solvent that is at least substantially non-toxic and useful also as a pharmaceutically acceptable carrier in liquid dosage forms. The extraction process also produces a significantly higher yield from a single extraction than the state-of- the-art. The liquid dosage forms are produced directly from the extraction process without requiring removal of the extraction solvent, reducing complexity and cost of processing.

[0016] The Patent Application WO2016057839A1 entitled “Compositions and methods for increasing the bioavailability of one or more compounds ” discloses a highly bioavailable compositions and related methods of improving the bioavailability of one or more compounds. The compositions and methods disclosed herein may be employed to improve the bioavailability of poorly soluble or poorly bioavailable ingredients in a subject.

[0017] The Patent Application IN201641028193A entitled “An anti-inflammatory composition for the treatment of acute joint inflammation and a process for preparation thereof” discloses an anti-inflammatory composition comprising turmeric alcohol extract, beta caryophyllene, 3 -o-acetyl- 11 -keto-beta-bo swellic acid, extract obtained from Kaempferia galangal and turmeric water extract. The invention also describes a process of preparing the composition containing these extracts, which mainly involves extracting curcumin alcohol extract from turmeric, extracting gum resin from Boswellia serrata, AKBA is isolated from the extracted gum resin and complexed with phospholipid, beta-caryophyllene is isolated from Piper nigrum by steam distillation and subjected to fractionation for purification, flavonoids rich aqueous extract is extracted from Kaempferia galangal and turmeric water extract powder is obtained from Curcuma longa. The extracts are combined together to form a complete natural matrix, homogenized and subjected to spray drying to obtain a uniform mixture in a powdered form. The composition is useful as an anti-inflammatory agent in treatment of conditions such as acute joint pain and osteoarthritis.

[0018] The Patent Application IN202041031859A entitled “Method of synthesizing a turmeric based composition having enhanced bioavailability ” discloses a method of synthesizing a turmeric based composition wherein the composition has an enhanced bioavailability in blood. The composition is prepared by micronization and homogenization of the turmeric extract. The composition is a water dispersible liquid.

[0019] Even though there are formulations based on herbal constituents but very few aim to treat the root cause of the disease. However, Ayurvedic and homeopathic systems of medicine focus on healing the major cause of the ailment than symptomatic treatment and therefore exhibit fewer side effects with respect to allopathic medicines.

[0020] Hence, there is a need for a formulation with a specific set of phytoconstituents that exhibits the synergistic effect in mitigating the inflammation and pain thus to be effective for the treatment of arthritis. Herbal remedies form an alternative source to relieve symptoms in patients with arthritis as well as to address the drawbacks associated with existing treatment methods with allopathic approach.

Summary of the invention

[0021] The present invention discloses a formulation comprising the combination of phytoconstituents for the treatment of joint inflammation. The formulation is a combination of phytoconstituents, which exhibit synergistic effect in mitigating the pain and inflammation in patients with arthritis.

[0022] The formulation comprises the phytoconstituents such as turmeric, Indian frankincense, cinnamon, curcumin and ginger that are potent in isolation and in combination with other herbs exhibits improved bioactivity in mitigating the inflammation.

[0023] The formulation comprises combination of curcumin from Curcuma longa at a concentration in a range between 10%- 13% w/w, Boswellia serrata extract at a concentration in a range between 13%-15% w/w, cinnamon oil extract at a concentration in a range between 10%- 13% w/w, total gingerols from ginger extract at a concentration in a range between 5% -7% w/w, extract of turmeric at a concentration in a range between 27%-30% w/w. [0024] The formulation is prepared using a unique combination of polar and nonpolar matrix to enhance the bioactivity of the individual ingredients. The phytoconstituents used in the formulation of the present invention are extracted and isolated from the sources using various processing methods to retain the bioactivity of the phytoconstituents.

[0025] Cinnamon oil extract is a non-polar entity and forms a non-polar matrix during homogenization. This matrix protects the turmeric-boswellic acid-ginger extract complex within its network from the digestive enzymes present in the gastro-intestinal tract. The lipophilic nature of the non-polar matrix also aids in easy absorption of the complex from the epithelial lining of the gastrointestinal tract.

[0026] The formulation with a polar non-polar complex increases the bioavailability thus enhancing the bioactivity of the constituents. The complex synergistic interactions between the ingredients enhance the bioavailability of active components, promote therapeutic effects, and reduce toxicity.

[0027] The formulation of the present invention is effective in reducing mono- iodoacetate-induced osteoarthritis in Wistar rats. The results indicated that the administration of the formulation of the present invention did not result in mortality or morbidity, does not alter any clinical signs or body weight in comparison with the control group. The formulation exhibited increase in grip strength and decreased the knee diameter in comparison to the induction control group at the end of 30 days. The formulation also decreased the levels of interleukin- ip (IL-1 P), tumor necrosis factor-a (TNF- a) and C-reactive Protein (CRP) indicating that the formulation of the present invention is effective in inhibiting mono-iodoacetate- induced osteoarthritis in Wistar rats.

[0028] The formulation exhibits improved bio -efficacy through the unique delivery mechanism by active penetration and increased stability of phytochemicals due to reduced degradation of phytochemicals.

Brief description of the drawings [0029] The foregoing and other features of embodiments will become more apparent from the following detailed description of embodiments when read in conjunction with the accompanying drawings. In the drawings, like reference numerals refer to like elements.

[0030] FIGURE 1 tabulates the composition of the formulation according to an embodiment of the invention.

[0031] FIGURE 2a illustrates the top view of the matrix embedded with the complex of the formulation.

[0032] FIGURE 2b illustrates the 3D view of the matrix embedded with the complex of the formulation.

[0033] FIG 3 tabulates the effect of the formulation on locomotor activity in Wistar rats.

[0034] FIG 3a and FIG 3b illustrates the effect of the formulation on the grip strength and the knee diameter in Wistar rats.

[0035] FIG 4 tabulates the effect of the formulation on the biochemical parameters in Wistar rats.

[0036] FIG 4a, 4b and FIG 4c illustrates the effect of the formulation on the biochemical parameters in Wistar rats.

Detailed description of the invention

[0037] In order to more clearly and concisely describe and point out the subject matter of the claimed invention, the following definitions are provided for specific terms, which are used in the following written description. [0038] The term "BioavailabiHly" refers to ability of a drug or other substance to be absorbed and used by the body.

[0039] The term “ Joint Inflammation" refers to a part of the process by which the immune system defends the body from harmful agents, such as bacteria and viruses. [0040] The term “ Phytoconstituent" refers to non-nutrient active plant chemical compounds or bioactive compounds and are responsible for protecting the plant against infections, infestations, or predation by microbes, pests, pathogens, or predators.

[0041] The present invention overcomes the drawback of the existing state of the art technologies by providing a formulation of combination of phytoconstituents for treatment of joint inflammation.

[0042] The invention discloses a formulation of phytoconstituents for the treatment of joint inflammation. The formulation comprises a combination of phytoconstituents, which exhibit synergistic effect in mitigating the pain and inflammation in patients with arthritis.

[0043] The formulation of the present invention comprises the phytoconstituents such as turmeric, Indian frankincense, cinnamon, curcumin and ginger that are potent in isolation and in combination with other herbs exhibits improved bioactivity.

[0044] FIG 1 tabulates the composition of the formulation according to an embodiment of the invention. The formulation comprises combination of curcumin from Curcuma longa at a concentration in a range between 10%-13% w/w, Boswellia serrata extract at a concentration in a range between 13%-15% w/w, cinnamon oil extract at a concentration in a range between 10%- 13% w/w, total gingerols from ginger extract at a concentration in a range between 5%-7% w/w, extract of turmeric at a concentration in a range between 27%-30% w/w. [0045] Curcumin used in the formulation is extracted and isolated from Curcuma longa by High-Performance Liquid Chromatography (HPLC) and turmeric is used as an aqueous extract. Curcumin, a polyphenol extract of Curcuma longa exhibits anti-inflammatory and antioxidant effects. The anti-inflammatory activity of curcumin is attributed to multiple mechanisms including the inhibition of the production of inflammatory mediators, such as interleukin (IL)-l, tumour necrosis factor-alpha (TNF-a), IL-8, Nitric Oxide (NO), and a variety of matrix metalloproteinase (MMPs) by inhibiting the activation of NF-KB, protein kinase B (Akt), and MAPK signaling pathways.

[0046] Turmeric from Curcuma longa exhibits antiarthritic effects by inhibiting joint inflammation and specifically joint destruction.

[0047] Boswellia serrata mainly comprises four pentacyclic triterpenic acids called as boswellic acids among several other terpenoids and are capable of inhibiting the synthesis of pro-inflammatory enzymes, microsomal prostaglandin E2 (PGE2) synthase- 1 and 5-lipoxygenase, reducing production or activation of inflammatory mediators such as matrix metalloproteinase (MMP)-9, MMP-13, cyclooxygenase (COX)-2, and nitric oxide (NO). In addition, boswellic acids exhibit analgesic and anti- arthritic effects. The presence of Boswellia exhibits therapeutic effects in arthritis by improving the knee joint gap, reducing osteophytes and attenuating inflammatory mediators such as C-reactive protein and hyaluronic acid that are associated with knee osteoarthritis.

[0048] Cinnamon oil extract from Cinnamon is commonly used in herbal formulations used in the treatment of inflammation. Cinnamon oil mainly comprises cinnamaldehyde, which exhibits anti-inflammatory and anti-rheumatic effects. Cinnamon extract is useful in regulating the immune system, preventing and treating inflammation. Cinnamon extract and its polyphenols are also associated with the reduction of serum levels of TNF-alpha, C-reactive protein (CRP), and interleukin (IL)-6. Cinnamon usually comprises multiple nutrients such as fiber, calcium, iron, magnesium, phosphorus, zinc, vitamin D, potassium etc. [0049] Gingerols from ginger extract isolated from rhizomes of the Zingiber officinale comprises diarylheptanoid, yakuchinone A, proanthocyanidine, 10- shogoal, 10-gingerol and exhibits anti-inflammatory and analgesic activities by inhibiting prostaglandin and leukotriene biosynthesis. Diarylheptanoid with catechol group exhibits activity against 5-lipoxygenase, which further inhibits leukotriene biosynthesis thus producing anti-inflammatory effect. Yakuchinone A inhibits prostaglandin production, which results in an anti-inflammatory effect.

[0050] In addition, 6-Gingerol, 6-shogoal, 8-shogaol and gingerdione also inhibits COXI and COX 2, PGE2 and NO synthesis. Gingerols exhibits additive joint protective effects in arthritis.

[0051] The ingredient of the formulation is prepared in the form of a matrix to enhance the bioactivity of the individual ingredients. The combination of all said ingredients forms a natural matrix and the combination of curcumin with Boswellic acid significantly improves physical performances in patients with arthritis when compared to patients administered with curcumin alone.

[0052] The phytoconstituents used in the formulation of the present invention are extracted and isolated from the respective sources using different methods to retain the bioactivity of the phytoconstituents.

[0053] The ingredient of the formulation exhibits synergistic effect by regulating the biomolecules that are involved in the mechanism thus mitigating the pain and inflammation to exhibit the efficacy. The complex synergistic interactions between the ingredients enhance the bioavailability of active components, promote therapeutic effects, and reduce toxicity.

[0054] The ingredients of the formulation form a natural matrix comprising polar and non-polar matrices. [0055] FIG 2 illustrates the matrix embedded with the formulation of the present invention. FIG 2a illustrates the top view of the matrix embedded with the complex of the formulation and FIG 2b illustrates the 3D view of the matrix embedded with the complex of the formulation. Cinnamon oil extract is a non-polar entity and forms a non-polar matrix during homogenization. This matrix protects the turmeric - boswellic acid-ginger extract complex within its network from the digestive enzymes present in the gastro-intestinal tract. The lipophilic nature of the non-polar matrix also aids in easy absorption of the complex from the epithelial lining of the gastrointestinal tract. The polar non-polar complex increases the bioavailability thus enhancing the bioactivity of the constituents.

[0056] The formulation of the present invention is prepared by mixing all the ingredients at specified concentration to form a natural matrix. The matrix is subjected to homogenization and spray drying to form a dry powdered mass. The powdered formulation is prepared as capsule by filling. The method of spray drying results in the formation of consistent, fine, optimum particle size of the powder. The capsule is generally prepared at a dosage of 250 mg and recommended for the treatment of joint inflammation in patients with arthritis.

[0057] The following examples are offered to illustrate various aspects of the invention. However, the examples are not intended to limit or define the scope of the invention in any manner.

Example 1: Evaluation of the efficacy of the formulation in rats

[0058] The efficacy of the formulation of the present invention is evaluated in mono-iodoacetate-induced osteoarthritis in Wistar rats.

[0059] The Wistar rats are classified into four groups namely normal control (Gl), induction control (G2), formulation group (G3) and reference group (G4). Osteoarthritis is induced by administering mono-iodoacetate. The injection is prepared by dissolving 3 mg of mono-iodoacetate in 50 pL of sterile saline. The normal control is orally treated with Placebo at a dose of 51.66 mg/kg body weight, formulation group is orally administered with a dose of 51.66 mg/kg body weight of the formulation of the present formulation and the reference group is orally administered with a dose of 51.66 mg/kg body weight of diclofenac sodium.

[0060] The Wistar days are observed for clinical signs, morbidity, mortality, body weight and locomotor activity parameters such as grip strength and knee diameter for 30 days. The biochemical parameters such as interleukin- ip (IL-1 P), tumor necrosis factor-a (TNF- a) and C-reactive Protein (CRP) are analyzed in the formulation group and reference group.

[0061] The results indicated that the administration of the formulation of the present invention did not result in mortality or morbidity, does not alter any clinical signs or body weight in comparison with the control group.

[0062] FIG 3 tabulates the effect of the formulation on locomotor activity in Wistar rats. The locomotor activity in Wistar rats is evaluated by analyzing the grip strength and the knee diameter. The administration of the formulation and the reference drug exhibited increase in grip strength and decreased the knee diameter in comparison to the induction control group at the end of 30 days.

[0063] FIG 3a and FIG 3b illustrates the effect of the formulation on the grip strength and the knee diameter in Wistar rats. The formulation exhibited improved grip strength and the knee diameter in Wistar rats.

[0064] FIG 4 tabulates the effect of the formulation on the biochemical parameters in Wistar rats. The results indicated that the levels of interleukin- ip (IL-1 P), tumor necrosis factor-a (TNF- a) and C-reactive Protein (CRP) are decreased in the treatment group. The results indicated that the formulation of the present invention is effective in inhibiting mono-iodoacetate-induced osteoarthritis in Wistar rats. [0065] FIG 4a, 4b and FIG 4c illustrates the effect of the formulation on the biochemical parameters in Wistar rats. The formulation decreased levels of interleukin- ip (IL-1 P), tumor necrosis factor-a (TNF- a) and C-reactive Protein (CRP) in Wistar rats. The results indicated that the formulation of the present invention is effective in inhibiting mono-iodoacetate-induced osteoarthritis in Wistar rats.

[0066] The formulation of the present invention does not affect the body weight, histological parameters and hence is safe and effective in reducing the mono- iodoacetate-induced osteoarthritis in Wistar rats.

[0067] The formulation exhibits improved bioavailability of the phytoconstituents due to the presence of Boswellia serrata and the cinnamon oil in the formulation.

[0068] The formulation of the present invention with a combination of specific herbal ingredients is safe, effective not only for symptomatic relief but also provide total relief in patients with arthritis. The formulation exhibits improved bio-efficacy through the delivery mechanism by active penetration and increased stability of phytochemicals due to reduced degradation of phytochemicals.