KR102566287B1 - Composition for synovial fluid substitute comprising hyaluronic acid - Google Patents

Abstract

The present invention relates to a composition for joint synovial fluid supplement containing hyaluronic acid, which suppresses the decrease in molecular weight of hyaluronic acid by satisfying specific conditions in a frequency sweep test and a rate sweep test, It maintains viscoelasticity to increase the stability of the formulation, and satisfies the osmotic pressure and pH range similar to that of normal synovial fluid, which can significantly alleviate pain that may occur in patients, thereby replacing synovial fluid lost during arthroscopic surgery and reducing friction in the joint area. It is useful for rapid functional recovery due to pain, reduction of the risk of infection at the joint surgery site, and pain relief for patients.

Description

Composition for synovial fluid substitute comprising hyaluronic acid

The present invention relates to a composition for synovial fluid supplementation containing hyaluronic acid, and more particularly, by satisfying specific conditions during a frequency sweep test and a rate sweep test while containing hyaluronic acid, The present invention relates to a composition for synovial fluid supplementation with high average recovery rate and small deformation, high stability of the composition for synovial fluid supplementation in terms of molecular weight and viscoelasticity of hyaluronic acid, and high pain relieving effect.

Recently, arthroscopic surgery is frequently performed as a method for treating joint lesions such as osteoarthritis or synovitis.

Arthroscopic surgery means making a hole through a minimal incision of less than 1 cm in diameter around the joint, inserting an endoscope with a small camera into the joint, and performing surgery while watching a high-resolution monitor. Types of such arthroscopic surgery include tendon and ligament reconstruction and suture related surgery, meniscus resection and suture related surgery, limb joint resection, vitrectomy and cruciate ligamentoplasty. As of 2016, these arthroscopic surgeries Approximately 330,000 patients received arthroscopic surgery, and many patients are undergoing arthroscopic surgery (HIRA, health care big data medical open system, treatment behavior statistics). On the other hand, the knee joint is the largest joint in the human body, and is known as the most suitable joint for performing arthroscopic surgery because it is not surrounded by muscles in the front and has no important blood vessels or nerves. Therefore, among arthroscopic surgeries, arthroscopy performed on the knee joint is most frequently performed, and it is known that about 80% of the total arthroscopic surgery is performed (Ahn Jin-hwan, orthopedic surgeon, Samsung Seoul Hospital, Korean doctor). association magazine). The names of arthroscopy performed on the knee joint include partial or total meniscectomy, meniscal repair, removal of loose body, synovectomy, and dissected bone. These include removal or fixation of osteochondritis dissecans, repair or reconstruction of cruciate ligament, and fracture fixation.

However, during such arthroscopic surgery, there is a risk of joint dysfunction due to generation of small fragments of cartilage or loss of synovial fluid. In addition, although physiological saline is generally used to wash the joint cavity after arthroscopic surgery, there is a problem in that the possibility of side effects increases because the physicochemical properties are different from synovial fluid. Therefore, a synovial fluid supplement made of a polymer solution such as hyaluronic acid is used to relieve pain by washing physiological saline used in arthroscopic surgery and temporarily replenishing and replacing lost synovial fluid.

Synovial fluid supplement is classified as a grade 4 biomaterial (medical device) for tissue repair, and is a product developed to replace synovial fluid lost after arthroscopic surgery. It is injected into the joint cavity after surgery to reduce friction in the joint. It reduces the risk of infection by quickly restoring functionality and coating the synovial cell layer of articular cartilage. Regarding this, only Viscoseal®, a hyaluronic acid-based isotonic solution that was approved for import by the Ministry of Food and Drug Safety in April 2016 as the first synovial fluid substitute in Korea, is being used. Viscosil forms a layer of hyaluronic acid while being coated with an isotonic hyaluronic acid solution. As this layer is activated, a masking effect on the joint occurs, reducing the pain caused by this and reducing the discomfort felt by the patient after joint surgery. It was developed with the purpose of On the other hand, there are currently no domestically produced products that have been licensed for manufacturing, but it is estimated that there is a market size of about 40 to 60 billion won.

As such, compared to the market size, there is still a lack of effective and stable development of synovial fluid supplements. In particular, since the initial pain relief of patients undergoing arthroscopic surgery is affected by the molecular weight and viscoelasticity of the polymer solution used as a synovial fluid supplement, a technique for improving stability in which the molecular weight and viscoelasticity of the polymer solution is stably maintained is important. In addition, since it temporarily replaces synovial fluid, it is important to satisfy the osmotic pressure and pH range similar to that of normal synovial fluid.

The present invention has been proposed to solve the above problems, and has a specific numerical range in the frequency sweep test and the shear rate change test (rate sweep test), whereby the hyaluronic acid molecular weight in the hyaluronic acid solution and the solution It is an object of the present invention to provide compositions for synovial fluid supplements that are stably maintained without decreasing viscoelasticity and have high stability and high pain relieving effect when injected into the human body.

The present invention has been made to solve the problems of the prior art as described above, and the storage modulus and phase angle measured according to the frequency change test, and the relaxation time measured according to the shear rate change test When the relaxation time and the shear thinning index are within a specific range, the molecular weight of the hyaluronic acid in the hyaluronic acid solution and the viscoelasticity of the solution are not lost and are stably maintained, resulting in high stability and pain relieving effect upon injection into the human body. It was confirmed that a composition for a high synovial fluid supplement can be provided and the present invention was completed.

As one aspect for achieving the above objects, the present invention includes hyaluronic acid or a salt thereof, and mannitol, and measures the storage modulus and phase angle measured according to the frequency change test, In addition, the relaxation time and shear thinning index measured according to the shear rate change test are in a specific range, specifically the storage modulus measured at 1 Hz according to the frequency change test is 3 Pa or more The phase angle may be 60° or less, or the storage modulus measured at 2.5 Hz may be 6 Pa or more and the phase angle may be 50° or less.

As another additional aspect, the synovial fluid supplement composition according to the present invention has a relaxation time of 0.1 seconds or more and a shear thinning index of 0.35 or less measured according to the shear rate change test Characterized in, it relates to a composition for joint synovial fluid supplement containing hyaluronic acid. Preferably, the composition for joint synovial fluid supplement is injected into a joint or a joint periphery, and replaces synovial fluid lost during arthroscopic surgery by replenishing synovial fluid, rapidly recovering functionality due to reduced friction in the joint, and reducing the risk of infection in the joint surgical site. , or for relieving joint pain in patients.

Hereinafter, the present invention will be described in detail.

Hyaluronic acid (hereinafter also referred to as 'HA') included in the synovial fluid supplement composition of the present invention is a biopolymer material in which repeating units composed of N-acetyl-D-glucosamine and D-glucuronic acid are linearly linked, It is abundant in the vitreous humor of the eyeball, synovial fluid of the joint, chicken comb, etc., and has excellent biocompatibility, so it is used for medical and medical devices such as ophthalmic surgery aids, joint function improvers, drug delivery materials, eye drops, and anti-wrinkle agents, as well as cosmetics. It is widely used.

Specifically, the hyaluronic acid included in the synovial fluid supplement composition of the present invention may include a salt thereof in addition to hyaluronic acid. The salt of hyaluronic acid includes, for example, inorganic salts such as sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, and cobalt hyaluronate, and organic salts such as tetrabutylammonium hyaluronate. being, but not limited thereto.

In the present invention, the average molecular weight of hyaluronic acid used as a synovial fluid supplement composition may be 1,800,000 Da or more, preferably 1,800,000 to 4,00,000 Da, and more preferably 2,500,000 to 3,500,000 Da.

In addition, the hyaluronic acid synovial fluid supplement composition according to the present invention may contain 0.1 to 3.0% by weight, preferably 0.3 to 1.0% by weight of hyaluronic acid, based on the total weight of the synovial fluid supplement composition.

In addition, the synovial fluid supplement composition according to the present invention contains mannitol. The mannitol may be included as a stabilizer for stabilizing hyaluronic acid included in the synovial fluid supplement. Unfortunately, conventional hyaluronic acid-containing compositions that can be injected into the human body by injection or the like partially or completely degrade prior to injection, particularly during sterilization steps such as high-temperature sterilization and/or when stored for any reasonably long period of time. have a tendency to become However, when mannitol is included in the synovial fluid supplement according to the present invention, even if the high-molecular hyaluronic acid is used, the reduction in molecular weight due to sterilization or storage as described above can be minimized to provide a stable synovial fluid supplement You can do it.

Preferably, the content of mannitol included in the synovial fluid supplement composition according to the present invention is 3% by weight or more, preferably 3% by weight or more and 10% by weight or less, more preferably 3% by weight or more based on the total weight of the synovial fluid supplement. It may be included in 7% by weight or less.

The hyaluronic acid-based synovial fluid supplement composition according to the present invention exhibits excellent stability in terms of molecular weight and viscoelasticity of hyaluronic acid, and specifically, according to a frequency change test, the storage modulus measured at 1 Hz is 3 Pa or more and the phase angle is 60 ° Or less, or characterized in that the storage modulus measured at 2.5 Hz is 6 Pa or more and the phase angle is 50 ° or less. Preferably, the storage modulus measured at 1Hz may be 2Pa or more to 20Pa or less, more preferably 3Pa or more to 15Pa or less, and the storage modulus measured at 2.5Hz is 5Pa or more to 25Pa or less, more preferably 6Pa It may be greater than or equal to 20Pa, and the measured phase angle at 1Hz may be greater than or equal to 20° and less than or equal to 60°, more preferably greater than or equal to 40° and less than or equal to 60°, and the measured phase angle at 2.5Hz may be greater than or equal to 20° to less than or equal to 60°. It may be 50° or less, more preferably 30° or more to 50° or less.

In the present invention, the frequency sweep test (oscillation) is to determine the change in rheological properties according to the shear rate in order to measure the viscoelastic properties according to the change in frequency, shear storage modulus (G ', storage modulus), shear Loss modulus (G ", loss modulus), phase angle (δ, phase angle), etc. can be measured. Specifically, the values according to the frequency change test according to the present invention are measured at 1 Hz and 2.5 Hz, and 1 Hz is The walking situation, 2.5Hz is defined as the frequency corresponding to the running situation.

As a specific aspect, the frequency change test is performed by loading the sample into a circular parallel plate geometry at room temperature, increasing the frequency from 0.01 to 100 Hz, and measuring rheological properties according to shear in the linear region. .

In addition, the shear rate change test (rate sweep test, roatation) is to confirm the change in rheological properties such as viscosity according to the change in shear rate. ), shear thinning index, etc. can be calculated. As a specific embodiment, the shear rate change test may be performed by loading the sample into a circular parallel plate geometry at room temperature and measuring the viscosity by increasing the shear rate from 0.01 to 500/s.

On the other hand, the synovial fluid supplement composition according to the present invention may further include a buffer, an isotonic agent, or a combination thereof in addition to the hyaluronic acid and mannitol. In a specific example, when the synovial fluid supplement composition further includes a buffer or a buffer and an isotonic agent, mannitol and/or an isotonic agent may be mixed with the buffer and mixed with hyaluronic acid or a salt thereof.

The buffer may be used without limitation as long as it is used for the preparation of hyaluronic acid. Examples of such preferred buffers are citric acid, sodium monohydrogen phosphate, sodium dihydrogen phosphate, acetic acid, diethyl barbituric acid, sodium acetate, TAPS (tris(hydroxymethyl)methylamino)propane Sulfonic Acid), Bicine (2-bis(2-hydroxyethyl)amino)acetic acid), Tris(tris(hydroxymethyl)ammoniummethane), Tricine(N-(2-hydroxy-1,1-bis(hydroxyl) methyl)ethyl)glycine), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), TES (2-[[1,3-dihydroxy-2-(hydroxymethyl)propane- 2-yl]amino]methanesulfonic acid) and PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid)), and buffer solutions containing one or more selected from the group consisting of, but are not limited thereto. The content of the components included in the buffer may be appropriately adjusted, but may be preferably included in a concentration of 0.03 to 2.0 g/L with respect to the buffer.

The isotonic agent may be used without limitation as long as it is used in the preparation of hyaluronic acid, and may be included in a buffer solution. As a preferred tonicity agent, sodium chloride may be used, but is not limited thereto. The content of the tonicity agent may be appropriately adjusted as needed, and may be included, for example, 0.1 to 9.0 g/L relative to the buffer, but is not limited thereto.

As an additional aspect, the composition according to the present invention may further include acceptable components that may be included in the preparation of the synovial fluid supplement composition in addition to the above components. Specifically, the composition may include one or more components selected from the group consisting of non-steroidal anti-inflammatory drugs, anesthetics, opioid analgesics, corticosteroids, antineoplastic agents, monoclonal antibodies, chimeric monoclonal antibodies, vitamins and minerals. . Examples of such ingredients include non-steroidal anti-inflammatory drugs (NSAIDS) (eg diclofenac, ibuprofen, piroxicam); anesthetics (eg lidocaine and bupivacaine); opioid analgesics (eg codeine and morphine); corticosteroids (eg dexamethasone and prednisone); anti-neoplastic agents (eg methotrexate); antiviral agents (eg acyclovir and vidarabine); may contain additional active or inactive ingredients including monoclonal antibodies (eg Humira) and chimeric monoclonal antibodies (eg infliximab).

The synovial fluid supplement composition according to the present invention may be formulated into an appropriate dosage form known in the art and administered to the joint or the periphery of the joint. The synovial fluid supplement composition according to the present invention may be administered in a dosage form suitable for such use, and preferably may be an injection.

Specifically, the administration method may include single or multiple intra-articular injections or implantation of the supplement in an amount sufficient to provide the desired physiological effect of the composition for lubricant supplement prepared according to the present invention. Intra-articular injection or implantation is performed directly into a bone and/or cartilage defect of a human or non-human mammal, eg by arthroscopy or by an injection device such as a syringe. Examples of administration sites include the knee, shoulder, ankle, temporomandibular and carpometacarpal joints, elbow, hip, wrist, and spinal discs.

As a further aspect, the present invention provides a pre-filled disposable syringe (pre-filled syringe) having a single dose of the synovial fluid supplement composition.

In addition, the present invention relates to a composition for relieving or treating pain and discomfort associated with joint disorders, joint damage including osteoarthritis and traumatic cartilage lesions, or joint diseases, including the synovial fluid supplement composition. Examples of such joint diseases include osteoarthritis (primary (idiopathic) or secondary), rheumatoid arthritis, joint injury (eg traumatic or repetitive motion injury), cartilage pathology (eg chondrocalconsis), cartilage maraxosis, and septic arthritis.

The hyaluronic acid-based synovial fluid supplement composition according to the present invention includes a specific range of mannitol as a stabilizer, and at the same time, the storage modulus and phase angle measured according to the characteristic frequency change test, and also Due to the relaxation time and shear thinning index calculated according to the shear rate change test, the decrease in molecular weight that can occur when sterilizing high molecular weight hyaluronic acid is suppressed and viscoelasticity is maintained As a synovial fluid supplement composition used not only for arthroscopic surgery but also for patients with arthritis, it not only enhances the stability of the formulation, but also satisfies the osmotic pressure and pH range similar to that of normal synovial fluid, thereby significantly reducing pain that may occur in patients. useful.

As another aspect, the present invention,

Performing a frequency change test and a shear rate change test on the composition for hyaluronic acid synovial fluid supplement; and

As a result of the frequency change test, the storage modulus measured at 1 Hz was 3 Pa or more, the phase angle was 60 ° or less, the storage modulus measured at 2.5 Hz was 6 Pa or more, and the phase angle was 50 ° or less, and whether the relaxation time measured according to the shear rate change test is 0.1 second or more, and the shear thinning index is 0.35 or less. And it provides a method for selecting a composition for synovial fluid supplementation with high pain relief.

The frequency change test and the shear rate change test may be performed according to known frequency change test and shear rate change test methods. As an example of each of these tests, the items described in the synovial fluid supplement composition may be applied as they are. In the case of a synovial fluid supplement composition having a storage modulus, phase angle, relaxation time, and shear thinning index within the applicable range, even if it contains high-molecular weight hyaluronic acid, it is unlikely to decrease molecular weight and viscoelasticity due to sterilization, etc., resulting in high stability and problems such as pain It is possible to clearly select the composition for synovial fluid replenishment in which is resolved.

The synovial fluid supplement composition according to the present invention includes hyaluronic acid, a salt thereof, or mannitol, and the frequency change test and the shear rate change test are the specific storage modulus and phase angle by the known frequency change test and shear rate change test method, In addition, by having a relaxation time and a shear thinning index, the decrease in hyaluronic acid molecular weight is suppressed and viscoelasticity is maintained to increase the stability of the formulation, and it satisfies the osmotic pressure and pH range similar to that of normal synovial fluid, thereby reducing pain that may occur to patients It is useful for replacing synovial fluid lost during arthroscopic surgery, rapidly recovering functionality by reducing friction in the joint, reducing the risk of infection in the joint surgical site, and relieving patient pain.

1 is a graph showing the measurement of the storage modulus and the phase angle as a result of a frequency change test of 1Hz (walking situation) of the compositions for synovial fluid supplements of Examples 1 to 3 and Comparative Examples 1 to 6 according to the present invention.
Figure 2 is a graph showing the measurement of the storage modulus and the phase angle as a result of the frequency change test of 2.5Hz (jumping situation) of the compositions for synovial fluid supplements of Examples 1 to 3 and Comparative Examples 1 to 6 according to the present invention.
Figure 3 is a graph showing the relaxation time (s) and the shear thinning index calculated through the shear rate change test results of the compositions for synovial fluid supplements of Examples 1 to 3 and Comparative Examples 1 to 6 according to the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for explaining the present invention in more detail, and it is to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present application. It will be self-explanatory.

Example 1: Preparation of synovial fluid supplement composition containing high molecular hyaluronic acid and 3% by weight of mannitol

For the preparation of the synovial fluid supplement composition according to the present invention, the composition was obtained by performing the following manufacturing method.

0.5 g of hyaluronic acid (average molecular weight: about 2.5 to 3.5 million Da) was dissolved at a concentration of 0.5% by weight in a buffer solution containing 3% by weight of mannitol, 0.35% by weight of sodium chloride and 0.043% by weight of sodium hydrogen phosphate. did The completely melted mixture was filled in a plastic syringe and sterilized for 10 minutes using a high-temperature moist heat sterilizer to prepare a synovial fluid supplement composition containing hyaluronic acid and mannitol as a stabilizer.

Example 2: Preparation of synovial fluid supplement composition containing high molecular weight hyaluronic acid and 4% by weight of mannitol

0.5 g of hyaluronic acid (average molecular weight: about 2.5 to 3.5 million Da) was dissolved at a concentration of 0.5% by weight in a buffer solution containing 4% by weight of mannitol, 0.16% by weight of sodium chloride and 0.043% by weight of sodium hydrogen phosphate. did The completely melted mixture was filled in a plastic syringe and sterilized for 10 minutes using a high-temperature moist heat sterilizer to prepare a synovial fluid supplement composition containing hyaluronic acid and mannitol as a stabilizer.

Example 3: Preparation of synovial fluid supplement composition containing high molecular hyaluronic acid and 5% by weight of mannitol

0.5 g of hyaluronic acid (average molecular weight: about 2.5 to 3.5 million Da) was dissolved at a concentration of 0.5% by weight in a buffer solution containing 5% by weight of mannitol and 0.043% by weight of sodium hydrogen phosphate. The completely melted mixture was filled in a plastic syringe and sterilized for 10 minutes using a high-temperature moist heat sterilizer to prepare a synovial fluid supplement composition containing hyaluronic acid and mannitol as a stabilizer.

Comparative Example 1: Mannitol-free polymeric hyaluronic acid synovial fluid supplement composition

For comparison with the present invention, a synovial fluid supplement composition was prepared by the following method.

0.5 g of hyaluronic acid (average molecular weight: about 2.5 to 3.5 million Da) was dissolved at a concentration of 0.5% by weight in a buffer solution containing 0.9% by weight of sodium chloride and 0.043% by weight of sodium hydrogen phosphate. The completely melted mixture was filled in a plastic syringe and sterilized for 10 minutes using a high-temperature moist heat sterilizer to prepare a synovial fluid supplement composition containing hyaluronic acid and mannitol as a stabilizer.

Comparative Example 2: Viscosil®

Viscosil®, a product licensed as a synovial fluid supplement, was used as a comparative example.

Comparative Example 3: Mannitol-free low-molecular-weight hyaluronic acid synovial fluid supplement composition

0.5 g of hyaluronic acid (average molecular weight: about 1.2 to 1.8 million Da) was dissolved at a concentration of 0.5% by weight in a buffer solution containing 0.9% by weight of sodium chloride and 0.043% by weight of sodium hydrogen phosphate. The completely melted mixture was filled in a plastic syringe and sterilized for 10 minutes using a high-temperature moist heat sterilizer to prepare a synovial fluid supplement composition containing hyaluronic acid and mannitol as a stabilizer.

Comparative Example 4: Preparation of a synovial fluid supplement composition containing low molecular weight hyaluronic acid and 3% by weight of mannitol

0.5 g of hyaluronic acid (average molecular weight: about 1.2 to 1.8 million Da) was dissolved at a concentration of 0.5% by weight in a buffer solution containing 3% by weight of mannitol, 0.35% by weight of sodium chloride and 0.043% by weight of sodium hydrogen phosphate. did The completely melted mixture was filled in a plastic syringe and sterilized for 10 minutes using a high-temperature moist heat sterilizer to prepare a synovial fluid supplement composition containing hyaluronic acid and mannitol as a stabilizer.

Comparative Example 5: Preparation of a synovial fluid supplement composition containing low molecular weight hyaluronic acid and 4% by weight of mannitol

0.5 g of hyaluronic acid (average molecular weight: about 1.2 to 1.8 million Da) was dissolved at a concentration of 0.5% by weight in a buffer solution containing 4% by weight of mannitol, 0.16% by weight of sodium chloride and 0.043% by weight of sodium hydrogen phosphate. did The completely melted mixture was filled in a plastic syringe and sterilized for 10 minutes using a high-temperature moist heat sterilizer to prepare a synovial fluid supplement composition containing hyaluronic acid and mannitol as a stabilizer.

Comparative Example 6: Preparation of a synovial fluid supplement composition containing low molecular weight hyaluronic acid and 5% by weight of mannitol

0.5 g of hyaluronic acid (average molecular weight: about 1.2 to 1.8 million Da) was dissolved at a concentration of 0.5% by weight in a buffer solution containing 5% by weight of mannitol and 0.043% by weight of sodium hydrogen phosphate. The completely melted mixture was filled in a plastic syringe and sterilized for 10 minutes using a high-temperature moist heat sterilizer to prepare a synovial fluid supplement composition containing hyaluronic acid and mannitol as a stabilizer.

Experimental Example 1: Frequency Sweep Test of hyaluronic acid-based synovial fluid supplement composition according to mannitol concentration (Frequency Sweep Test)

The sample was loaded into a 40 mm diameter circular parallel plate geometry of DHR (Discovery Hybrid Rhemeter, manufactured by TA), the frequency was increased from 0.01 to 100 Hz, and the strain in the linear region was 1 Hz and 2.5 Hz. The storage modulus and phase angle were measured at room temperature (25°C).

As the experimental group, Examples 1 to 3 containing the high-molecular weight hyaluronic acid with the content of mannitol at 3% by weight, 4% by weight and 5% by weight, respectively, relative to the total composition, and as a control group, the high-molecular weight hyaluronic acid without mannitol and Viscoseal®, a commercially available hyaluronic acid-based synovial fluid supplement, and low molecular weight hyaluronic acid without mannitol and mannitol at 3% by weight, 4% by weight, and 5% by weight of the total composition, respectively. Comparative Examples 1 to 6 including were used, and the results are shown in Table 1 and FIGS. 1 and 2 below.

division Frequency (1 Hz) Frequency (2.5 Hz) Storage modulus (G') phase angle (°) Storage modulus (G') phase angle (°) Example 1 3.40299 56.07 6.79024 47.6078 Example 2 4.33305 53.4818 8.24196 45.2989 Example 3 7.83133 46.2094 13.4505 38.4547 Comparative Example 1 1.83709 61.2421 4.19973 51.7535 Comparative Example 2 0.15575 80.7059 1.01239 64.3948 Comparative Example 3 0.07379 83.3535 0.41338 74.3663 Comparative Example 4 0.15115 80.0522 0.72926 69.3581 Comparative Example 5 0.2328 79.0694 0.92917 70.3577 Comparative Example 6 0.74069 73.2863 2.34768 63.9666

As can be seen in Table 1 and FIGS. 1 and 2, Examples 1 to 3, which are hyaluronic acid-based synovial fluid supplemental compositions containing high-molecular hyaluronic acid and mannitol at a concentration of 3% by weight or more relative to the total weight of the composition, are the control group. Comparative Example 1, a composition for synovial fluid replenishment based on hyaluronic acid containing high-molecular and low-molecular-weight hyaluronic acid that does not contain Viscosyl or mannitol, which are commercially available preparations, or low-molecular-weight hyaluronic acid and containing mannitol at a concentration of 3% by weight or more relative to the total weight of the composition to 6, the storage modulus (G′) is large and the phase angle is small at both 1 Hz and 2.5 Hz assuming walking and running situations. Therefore, it was confirmed that the synovial fluid supplement composition that satisfies the conditions of storage modulus and phase angle according to the frequency change test has high stability and a large pain relief effect.

Based on these results, in order to ensure stability and pain relieving effect upon injection of the synovial fluid composition containing hyaluronic acid and mannitol into the human body, in the test described above, the measured storage modulus at 1 Hz was 3 Pa or more and the phase angle was 60 ' Less than, preferably, the storage modulus measured at 2.5Hz is 6Pa or more and the phase angle is 50' or less.

Experimental Example 2: Shear rate change test of hyaluronic acid-based synovial fluid supplement composition according to mannitol concentration (Rate Sweep Test)

The sample was loaded into a 40 mm diameter circular parallel plate geometry of DHR (Discovery Hybrid Rhemeter, manufactured by TA), and the viscosity was measured at room temperature (25 ° C) while increasing the shear rate from 0.01 to 500 / s, The measured viscosity results were substituted into the Carreau-Yasuda Model to calculate the relaxation time and shear thinning index.

As the experimental group, Examples 1 to 3 containing the high-molecular weight hyaluronic acid with the content of mannitol at 3% by weight, 4% by weight and 5% by weight, respectively, relative to the total composition, and as a control group, the high-molecular weight hyaluronic acid without mannitol and Viscoseal®, a commercially available hyaluronic acid-based synovial fluid supplement, and low molecular weight hyaluronic acid without mannitol and mannitol at 3% by weight, 4% by weight, and 5% by weight of the total composition, respectively. Comparative Examples 1 to 6 including the above were used, and the results are shown in Table 2 and FIG. 3 below.

division Viscosity (Pa s) Relaxation time (s) Shear Thinning Index Example 1 2.01115 0.17717 0.29508 Example 2 2.5887 0.25456 0.31273 Example 3 5.86802 0.42181 0.24307 Comparative Example 1 0.95073 0.1627 0.42486 Comparative Example 2 0.17212 0.01036 0.3932 Comparative Example 3 0.10217 0.01272 0.60685 Comparative Example 4 0.15729 0.00661 0.36523 Comparative Example 5 0.22632 0.0077 0.35242 Comparative Example 6 0.51889 0.03761 0.42421

As can be seen in Table 2 and FIG. 3, Examples 1 to 3, which are hyaluronic acid-based synovial fluid supplemental compositions containing polymeric hyaluronic acid and mannitol at a concentration of 3% by weight or more relative to the total weight of the composition, are commercially available as controls. Comparative Examples 1 to 6, which are compositions for synovial fluid replenishment based on hyaluronic acid containing high-molecular and low-molecular-weight hyaluronic acid or low-molecular-weight hyaluronic acid that do not contain Viscosyl or mannitol, and mannitol at a concentration of 3% by weight or more relative to the total weight of the composition Compared to , it can be seen that the viscosity is high, the relaxation time is long, and the shear thinning index is small. Therefore, it was confirmed that the synovial fluid supplement composition that satisfies the conditions of relaxation time and shear thinning index according to the shear rate change test would have high stability and a large pain relief effect.

Based on these results, in order to ensure the safety and pain relief effect of the synovial fluid composition containing hyaluronic acid and mannitol when injected into the human body, it is necessary to show a relaxation time of 0.1 second or more and a shear thinning index of 0.35 or less in the test described above. desirable.

Claims (8)

It includes hyaluronic acid or a salt thereof and mannitol, wherein the mannitol is included in an amount of 3% by weight or more and 10% by weight or less based on the weight of the total composition,
The hyaluronic acid or its salt is included in 0.3 to 1.0% by weight based on the weight of the total composition,
According to the frequency change test, the storage modulus measured at 1Hz is 3 Pa or more and the phase angle is 60° or less, or the storage modulus measured at 2.5Hz is 6 Pa or more and the phase angle is 50° or less represents,
Characterized in that the relaxation time measured according to the shear rate change test is 0.1 to 0.42181 seconds and the shear thinning index is 0.24307 to 0.35, and the hyaluronic acid has an average molecular weight of 1,800,000 Da to 1,800,000 Da. A composition for a synovial fluid supplement comprising hyaluronic acid of 4,000,000 Da. delete delete delete The composition for synovial fluid supplement according to claim 1, further comprising a buffer, an isotonic agent, or a combination thereof. A composition for treating arthritis or relieving joint pain, comprising the composition for synovial fluid supplement according to claim 1 or 5. A pre-filled syringe containing the composition for a synovial fluid supplement according to claim 1 or 5. Performing a frequency change test or a shear rate change test on the composition for hyaluronic acid synovial fluid supplement; and
According to the frequency change test, the storage modulus measured at 1 Hz is 3 Pa or more and the phase angle is 60 ° or less, or the storage modulus measured at 2.5 Hz is 6 Pa or more and the phase angle is 50 ° whether it indicates the following; or
It is characterized in that the relaxation time calculated according to the shear rate change test is 0.1 to 0.42181 seconds and the shear thinning index is 0.24307 to 0.35, wherein the hyaluronic acid synovial fluid supplement composition contains hyaluronic acid or a salt thereof and mannitol, wherein the mannitol is included in an amount of 3% by weight or more and 10% by weight or less based on the weight of the total composition;
The hyaluronic acid or its salt is included in 0.3 to 1.0% by weight based on the weight of the total composition,
The method for selecting a composition for synovial fluid supplement, wherein the hyaluronic acid comprises hyaluronic acid having an average molecular weight of 1,800,000 Da to 4,000,000 Da.