JP2015078125A - Composition useful for pain treatment and method for treating pain using composition concerned - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a pharmaceutical composition useful for pain treatment which is a different type from conventional pain treatment.SOLUTION: The invention provides a pharmaceutical composition for treating diseases in patients comprising supernatant of a culture medium obtained by culturing tissues obtained from the patient.

Description

The present invention relates to a composition useful for treating pain and a method for treating pain using the composition.
More specifically, in one aspect, the present invention provides a culture in which cells constituting the patient's own (self) tissue contain the patient's own (self) serum for a patient exhibiting pain symptoms. The present invention relates to a method for treating pain using a physiologically active substance produced in a fluid.

  According to a survey of men and women over 20 years of age, one out of 4.4 adults in Japan has chronic pain. This means that about 23 million people in Japan have chronic pain.

  Disease names and symptoms causing chronic pain are low back pain (55.7%), forty shoulders / fifty shoulders, stiff shoulders (27.9%), and headache / migraine (20.7%). And about 70% of those who have chronic pain feel loss of motivation and mental stress.

  In the United States, the number of people with chronic pain is higher than the total number of people with heart disease, diabetes and cancer. In fact, in the United States, about 50 million people suffer from chronic pain, spending about $ 100 billion per year for treatment.

  Unfortunately, many of the strongest available analgesics have serious side effects. These side effects include risks such as addiction, addiction, increased heart attack and stroke.

  Moreover, many pain symptoms, particularly chronic pain symptoms, cannot be effectively treated with existing medications.

  CELEBREX® ($ 2.8 billion in 2004; GD Seale & Co., Skokie, Illinois, USA) and VIOXX® ($ 1.4 billion in 2004, Merck & Co., USA Needless to consider the revenue of drugs such as White House Station, New Jersey), effective treatment of pain symptoms, especially chronic pain symptoms, will undoubtedly be very helpful to human health.

  Thus, there is an unmet need for effective pain treatment.

  An object of the present invention is to provide a composition useful for the treatment of pain, which is a type different from the conventional treatment of pain.

  More specifically, in one aspect, the present invention provides, in a culture solution in which the patient's own (self) tissue constituent cells contain the patient's own (self) serum for a patient who exhibits pain symptoms. To treat pain using a physiologically active substance produced in Such treatment is superior in that it does not use drugs or chemical substances and does not use components derived from other people or other animals.

  In order to solve the above-described problems, the present invention has the following features.

<Pharmaceutical composition>
In one aspect, the pharmaceutical composition of the present invention is a pharmaceutical composition for treating a disease of a patient, comprising a culture solution obtained by culturing a tissue obtained from the patient. It is a thing.

  The culture solution obtained by culturing the tissue obtained from the patient can be used as it is in the pharmaceutical composition of the present invention, but can also be used after removing the cells. As a method for removing cells, for example, (1) a method of centrifuging the culture solution and collecting a supernatant after centrifugation of the culture solution, and / or (2) a cell or the like using a filter Can be removed. Preferably, the culture solution is centrifuged, and the supernatant after centrifugation of the culture solution is collected and then removed from the supernatant by further filtering cells and the like using a filter. it can.

  Here, the disease of the patient may be pain, particularly chronic pain or rheumatic pain.

  Moreover, culture | cultivation of the tissue acquired from the said patient can be performed in the culture solution which added the serum obtained from the said patient's autologous blood in one aspect | mode.

  Moreover, culture | cultivation of the tissue acquired from the said patient can be performed for 3 to 9 days in one aspect | mode.

  Moreover, the tissue acquired from the patient may be an adipose tissue in one aspect.

<Treatment method>
In one aspect, the method for treating a patient according to the present invention is a method comprising administering to the patient a pharmaceutical composition containing an effective amount of a culture solution obtained by culturing tissue obtained from the patient. .

  The culture solution obtained by culturing the tissue obtained from the patient can be used as it is in the treatment method of the present invention, but can also be used after removing the cells. As a method for removing cells, for example, (1) a method of centrifuging the culture solution and collecting a supernatant after centrifugation of the culture solution, and / or (2) a cell or the like using a filter Can be removed. Preferably, the culture solution is centrifuged, and the supernatant after centrifugation of the culture solution is collected and then removed from the supernatant by further filtering cells and the like using a filter. it can.

  Here, the disease of the patient may be pain, particularly chronic pain or rheumatic pain.

  Moreover, culture | cultivation of the tissue acquired from the said patient can be performed in the culture solution which added the serum obtained from the said patient's autologous blood in one aspect | mode.

  Moreover, culture | cultivation of the tissue acquired from the said patient can be performed for 3 to 9 days in one aspect | mode.

  Moreover, the tissue acquired from the patient may be an adipose tissue in one aspect.

<Manufacturing method>
In one aspect, the method for producing a pharmaceutical composition of the present invention is a method for producing a pharmaceutical composition for treating a disease of a patient, comprising: (1) culturing a tissue obtained from the patient; 2) A step of removing cells from a culture solution obtained by culturing.

  As a method for removing cells, for example, (1) a method of centrifuging the culture solution and collecting a supernatant after centrifugation of the culture solution, and / or (2) a cell or the like using a filter Can be removed. Preferably, the culture solution is centrifuged, and the supernatant after centrifugation of the culture solution is collected and then removed from the supernatant by further filtering cells and the like using a filter. it can.

  Here, the disease of the patient may be pain, particularly chronic pain or rheumatic pain.

  Moreover, culture | cultivation of the tissue acquired from the said patient can be performed in the culture solution which added the serum obtained from the said patient's autologous blood in one aspect | mode.

  Moreover, culture | cultivation of the tissue acquired from the said patient can be performed for 3 to 9 days in one aspect | mode.

  Moreover, the tissue acquired from the patient may be an adipose tissue in one aspect.

  The pharmaceutical composition of the present invention can be advantageously used for the treatment of pain (particularly chronic pain and rheumatic pain).

  The pharmaceutical composition of the present invention contains a culture solution obtained by culturing tissue obtained from the patient himself.

  The tissue obtained from the patient itself is preferably adipose tissue, but is not limited thereto.

  Moreover, there is no limitation in particular as a method of acquiring a structure | tissue from a patient itself. For example, liposuction can be used. As another method, the surgical wound can be cut out.

  The tissue obtained from the patient itself may be cultured by culturing the tissue itself, for example, by chopping cells with a scalpel and further culturing cells obtained by enzyme treatment.

  Culture of the tissue obtained from the patient himself can be performed by a well-known method. In one embodiment, it can be performed in a culture solution (eg, DMEM / F12 phenol red (−)) supplemented with serum obtained from the patient's own autologous blood.

  There is no particular limitation on the culture period of the tissue obtained from the patient himself. For example, it can be performed for 3 to 9 days.

  In the pharmaceutical composition of the present invention, a culture solution obtained by culturing a tissue obtained from a patient himself is used. The culture solution obtained by culturing the tissue obtained from the patient can be used as it is in the pharmaceutical composition of the present invention, but can also be used after removing the cells. As a method for removing cells, for example, (1) a method of centrifuging the culture solution and collecting a supernatant after centrifugation of the culture solution, and / or (2) a cell or the like using a filter Can be removed. Preferably, the culture solution is centrifuged, and the supernatant after centrifugation of the culture solution is collected and then removed from the supernatant by further filtering cells and the like using a filter. it can. In addition, the culture solution can be stored at a low temperature (for example, frozen or 4 ° C.) before use.

  The pharmaceutical composition of the present invention comprises the culture solution collected in this way. Such a culture solution is considered to contain various physiologically active substances produced when culturing a tissue (or a cell obtained by dissociation from the tissue) obtained from the patient itself. Therefore, the surprising and remarkable effect of the pharmaceutical composition of the present invention is that such various physiologically active substances are effective in the healing of organic and / or functional damage in the patient's body. Inferred.

  The pharmaceutical composition of the present invention is basically based on components derived from the patient himself and can be said to be particularly preferable from the viewpoint of side effects.

  There is no limitation on how to administer the pharmaceutical composition of the present invention to a patient. For example, a culture solution obtained by culturing a tissue obtained from the patient (including those obtained by removing cells from the culture solution, for example, a supernatant after centrifugation and a filtrate after filtration) It can be intravenously administered to a patient by drip infusion after being diluted with Lactec Ringer solution. It can also be administered to a patient without such dilution.

  There is no particular limitation on the frequency of administration of the pharmaceutical composition of the present invention to a patient. However, since the pharmaceutical composition of the present invention can exert a sufficient therapeutic effect even if administered once a month, it is preferable from the viewpoint of compliance.

The terms used in the present specification are used to describe specific embodiments, and are not intended to limit the invention.
In addition, the term “comprising” as used herein is intended to mean that there is a matter (member, step, element, number, etc.) described, unless the context clearly requires a different understanding. It does not exclude the presence of other items (members, steps, elements, numbers, etc.).
Unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms used herein should be interpreted as having a meaning consistent with the meaning in this specification and the related technical field, unless otherwise defined, idealized, or overly formal. It should not be interpreted in a general sense.
Although terms such as first, second, etc. may be used to represent various elements, it is understood that these elements should not be limited by those terms. These terms are only used to distinguish one element from another, for example, the first element is referred to as the second element, and similarly, the second element is the first element. Can be made without departing from the scope of the present invention.

  Hereinafter, the present invention will be described more specifically by way of examples. However, the present invention can be embodied in various forms and should not be construed as limited to the examples described herein. .

<Example 1> ● 1 ● Pain Syndrome case 1.36 years old, female, Ulnar nerve region pain

(1) Adipose tissue collection and sample preparation Approximately 10 cc of adipose tissue was aspirated into the abdominal adipose tissue and sucked into the syringe (Tsumcent method). The tissue was transferred to a petri dish and further minced with a scalpel. The minced tissue was transferred to a centrifuge tube, 25 mL of 0.5% trypsin solution was added, and enzyme treatment was performed at 37 ° C. for 30 minutes. Subsequently, the supernatant was removed by centrifugation at 3000 rpm for 10 minutes.
Cells precipitated in the centrifuge tube were resuspended in 30 mL of a culture solution (DMEM / F12 phenol red (−)) supplemented with 10% of serum obtained from autologous blood to obtain a suspension.
The suspension was transferred to two 75 cm 2 flasks for cell culture, and cultured in a culture apparatus at 37 ° C., 5% carbon dioxide gas, 100% humidity. On the third day of culture, the culture solution was transferred to a centrifuge tube, centrifuged at 3000 rpm for 10 minutes, and the supernatant was stored (4 ° C.). From the centrifuged supernatant, a filtrate was obtained using a filter having a pore size of 0.45 μm (hereinafter referred to as the third day culture supernatant). Cells are present on the bottom of the two flasks excluding the culture solution. To these two flasks, 15 mL each of the above culture solution supplemented with 10% serum obtained from autologous blood was added, and the culture was continued.
The culture was continued in the same manner to obtain culture supernatants on the 6th and 9th days (hereinafter referred to as the 6th and 9th day culture supernatants). Each culture supernatant was stored at 4 ° C.

(2) Administration method 20 mL of the culture supernatant was diluted in 500 mL of physiological saline for infusion. Subsequently, it was administered by infusion over 3 hours into the vein of the upper arm of the patient.

(3) Medical history The medical history of this patient is as follows.

(4) Therapeutic results The therapeutic effects after administration of the culture supernatant are shown below using VAS. Here, VAS means a visual analog scale. VAS is one method of scoring a patient's pain. In VAS, “maximum pain” is 10 and “no pain” is 0.

<Example 2> 2 ● Multiple rheumatoid arthritis case 1.80 years old, female, right femoral neck fracture, myocardial infarction, lumbar compression fracture, rheumatoid arthritis

(1) Adipose tissue collection and sample preparation Approximately 10 g of adipose tissue was cut out from the surgical wound edge during surgery for a femoral neck fracture. The tissue was transferred to a petri dish and further minced with a scalpel. The minced tissue was transferred to a centrifuge tube, 25 mL of 0.5% trypsin solution was added, and enzyme treatment was performed at 37 ° C. for 30 minutes. Subsequently, the supernatant was removed by centrifugation at 3000 rpm for 10 minutes.
Cells precipitated in the centrifuge tube were resuspended in 30 mL of a culture solution (DMEM / F12 phenol red (−)) supplemented with 10% of serum obtained from autologous blood to obtain a suspension.
The suspension was transferred to two 75 cm 2 flasks for cell culture, and cultured in a culture apparatus at 37 ° C., 5% carbon dioxide gas, 100% humidity. On the third day of culture, the culture solution was transferred to a centrifuge tube, centrifuged at 3000 rpm for 10 minutes, and the supernatant was stored (4 ° C.). From the centrifuged supernatant, a filtrate was obtained using a filter having a pore size of 0.45 μm (hereinafter referred to as the third day culture supernatant). Cells are present on the bottom of the two flasks excluding the culture solution. To these two flasks, 15 mL each of the above culture solution supplemented with 10% serum obtained from autologous blood was added, and the culture was continued.
The culture was continued in the same manner to obtain a culture supernatant on day 6 (hereinafter referred to as culture supernatant on day 6). Each culture supernatant was stored at 4 ° C.

(2) Administration method 20 mL of culture supernatant was diluted in 500 mL of lactec Ringer's solution. Subsequently, it was administered by infusion over 3 hours into the vein of the upper arm of the patient.

(3) Medical history The medical history of this patient is as follows. That is, this patient has undergone a femoral neck fracture joint on June 10, 2011.

(4) Therapeutic results The therapeutic effects after administration of the culture supernatant are shown below using VAS.

Case 2.73 years old, female, multiple rheumatoid arthritis, pyelonephritis, hypertension, glaucoma

(1) Adipose tissue collection and sample preparation About 1 cm square tissue was cut out from the abdominal wall adipose tissue. The tissue was transferred to a petri dish and further minced with a scalpel. The minced tissue was transferred to a centrifuge tube, 25 mL of 0.5% trypsin solution was added, and enzyme treatment was performed at 37 ° C. for 30 minutes. Subsequently, the supernatant was removed by centrifugation at 3000 rpm for 10 minutes.
Cells precipitated in the centrifuge tube were resuspended in 30 mL of a culture solution (DMEM / F12 phenol red (−)) supplemented with 10% of serum obtained from autologous blood to obtain a suspension.
The suspension was transferred to two 75 cm 2 flasks for cell culture, and cultured in a culture apparatus at 37 ° C., 5% carbon dioxide gas, 100% humidity. On the third day of culture, the culture solution was transferred to a centrifuge tube, centrifuged at 3000 rpm for 10 minutes, and the supernatant was stored (4 ° C.). A filtrate was obtained from the centrifuged supernatant using a filter having a pore size of 0.45 μm. (Hereinafter referred to as the third day culture supernatant.) Cells are present on the bottom of the two flasks excluding the culture solution. To these two flasks, 15 mL each of the above culture solution supplemented with 10% serum obtained from autologous blood was added, and the culture was continued.

(2) Administration method 20 mL of the culture supernatant was diluted in 500 mL of physiological saline for infusion. Subsequently, it was administered by infusion over 3 hours into the vein of the upper arm of the patient.

(3) Therapeutic results The therapeutic effects after administration of the culture supernatant are shown below.

Case 3. 70 years old, female, multiple rheumatoid arthritis, hypertension

(1) Adipose tissue collection and sample preparation About 10 cc of adipose tissue was aspirated from the abdominal wall adipose tissue into the syringe (Tsumcent method). The tissue was transferred to a petri dish and further minced with a scalpel. The minced tissue was transferred to a centrifuge tube, 25 mL of 0.5% trypsin solution was added, and enzyme treatment was performed at 37 ° C. for 30 minutes. Subsequently, the supernatant was removed by centrifugation at 3000 rpm for 10 minutes.
Cells precipitated in the centrifuge tube were resuspended in 30 mL of a culture solution (DMEM / F12 phenol red (−)) supplemented with 10% of serum obtained from autologous blood to obtain a suspension.
The suspension was transferred to two 75 cm 2 flasks for cell culture, and cultured in a culture apparatus at 37 ° C., 5% carbon dioxide gas, 100% humidity. On the third day of culture, the culture solution was transferred to a centrifuge tube, centrifuged at 3000 rpm for 10 minutes, and the supernatant was stored (4 ° C.). From the centrifuged supernatant, a filtrate was obtained using a filter having a pore size of 0.45 μm (hereinafter referred to as the third day culture supernatant). Cells are present on the bottom of the two flasks excluding the culture solution. To these two flasks, 15 mL each of the above culture solution supplemented with 10% serum obtained from autologous blood was added, and the culture was continued.

(2) Administration method 20 mL of culture supernatant was diluted in 500 mL of lactec Ringer's solution. Subsequently, the patient was intravenously infused into the vein of the upper arm over 3 hours.

(3) Therapeutic results The therapeutic effects after administration of the culture supernatant are shown below.

  Surprisingly, as described above, the pharmaceutical composition of the present invention, that is, the pharmaceutical composition containing a culture solution obtained by culturing a tissue obtained from a patient has pain (especially chronic pain and rheumatic pain). It turns out that there is a remarkable effect.

  The pharmaceutical composition of the present invention can be advantageously used for the treatment of pain (particularly chronic pain and rheumatic pain).

Claims (17)

A pharmaceutical composition for treating a disease in a patient comprising
A pharmaceutical composition comprising a culture solution obtained by culturing a tissue obtained from the patient. A pharmaceutical composition according to claim 1,
A pharmaceutical composition, wherein a culture solution obtained by culturing a tissue obtained from the patient is one from which cells have been removed. A pharmaceutical composition according to claim 2,
The pharmaceutical composition, wherein the cells are removed by centrifugation and / or filtration. A pharmaceutical composition according to claim 3,
The removal of the cells is performed by centrifuging the culture solution, collecting the supernatant after centrifugation of the culture solution, and then removing the cells and the like from the supernatant using a filter. The pharmaceutical composition characterized by being performed. A pharmaceutical composition according to any one of claims 1-4,
A pharmaceutical composition, wherein the disease of the patient is pain. A pharmaceutical composition according to any one of claims 1-4,
A pharmaceutical composition, wherein the patient's disease is chronic pain or rheumatic pain. A pharmaceutical composition according to any one of claims 1-4,
A pharmaceutical composition, wherein the tissue obtained from the patient is cultured in a culture solution supplemented with serum obtained from the patient's autologous blood. A pharmaceutical composition according to any one of claims 1-4,
A pharmaceutical composition, wherein culture of a tissue obtained from the patient is performed for 3 to 9 days. A pharmaceutical composition according to any one of claims 1-4,
A pharmaceutical composition, wherein the tissue obtained from the patient is a fat tissue. A method of treating a patient's disease,
A therapeutic method comprising administering to the patient a pharmaceutical composition comprising a culture solution obtained by culturing a tissue obtained from the patient in an effective amount. The treatment method according to claim 10, comprising:
A method of treatment, wherein the disease of the patient is pain. A method for producing a pharmaceutical composition for treating a disease in a patient comprising:
Culturing tissue obtained from the patient;
Removing cells from the culture medium obtained by culturing;
The manufacturing method characterized by including. It is a manufacturing method of Claim 12, Comprising:
The manufacturing method, wherein the step of culturing the tissue obtained from the patient is a step of culturing the tissue obtained from the patient in a culture solution to which serum obtained from the patient's autologous blood is added. It is a manufacturing method of Claim 13, Comprising:
The manufacturing method, wherein the step of removing the cells is performed by centrifugation and / or filtration. A pharmaceutical composition according to claim 14,
The step of removing the cells is performed by centrifuging the culture solution obtained in step 1 and collecting the supernatant after centrifugation of the culture solution, and further using the filter from the supernatant, A pharmaceutical composition comprising the step of removing others by filtration. It is a manufacturing method of Claim 12, Comprising:
The manufacturing method, wherein the step of culturing the tissue obtained from the patient is a step of culturing the tissue obtained from the patient for 3 to 9 days. It is a manufacturing method of Claim 12, Comprising:
The method for culturing tissue obtained from the patient is a step for culturing adipose tissue obtained from the patient.