JP6427039B2 - Evaluation method of physical pain during bipedal walking of non-human primates - Google Patents

Description

  The present invention provides a method for evaluating the intensity of body motion pain during biped walking of non-human primates from the walking state, and the presence or absence of the action of the test substance to alleviate body pain using the method, and It relates to a method of evaluating the action strength.

  Pain model animals are widely used to elucidate the mechanism of pain and develop analgesics. Animals, unlike humans, can not report the degree or condition of pain on their own. For this reason, in the pain test using an animal, the intensity of pain is evaluated by using the animal's behavior (pain-related behavior) to various stimuli causing pain. There are various types of pain, such as inflammatory pain, neuropathic pain, cancer pain, etc., and the mechanism is different, so various pain model animals are known. As inflammatory pain model animals, rats and mice which have been treated with chemical stimulants in their hind limbs are widely known. For example, a mouse in which arthritis is induced by injecting complete Freund's adjuvant (CFA) into a knee joint is used as a pain model animal of osteoarthritis (OA) (see Non-patent Document 1).

  Non-human primates are closer to humans than to rodents in terms of both genetics, neuroanatomical and pharmacokinetics, so they have an effect similar to that of human administration. It can be expected to be obtained. Therefore, in order to develop an analgesic that is effective for humans, it is preferable to use a non-human primate as a pain model animal rather than a rodent. However, since many non-human primates are originally four-legged animals, it is very difficult to analyze non-human primates for pain with a site of pain in the hind limb such as osteoarthritis of the knee. Have difficulty.

Westlund, et al. , Translational Research, 2012, vol. 160 (1), p. 84-94.

  The present invention provides a method for evaluating the intensity of body movement pain during bipedal walking of non-human primates, and the presence or absence of the action substance alleviating body movement pain of the test substance and the action intensity using the method. Aims to provide a way to Body pain is one of the indicators of pain.

  As a result of earnest research aimed at solving the above problems, the inventor of the present invention has evaluated the walking condition in a state of being installed in a walking aid that holds a posture during biped walking by the hind limbs. The inventors have found that it is possible to objectively evaluate body movement pain at the time of foot walking and complete the present invention.

That is, the method for evaluating body motion pain in bipedal walking of non-human primate according to the present invention, the method for evaluating action for relieving body pain of test substance, and substance having action for relieving body motion pain The screening method is as the following [1] to [8].
[1] A method for evaluating body pain in a test animal, wherein a non-human primate is the test animal, wherein the test animal is placed on a walking aid that holds a posture during biped walking by the hind limbs And a step of evaluating the walking state by freely walking only with the hind limbs in the stopped state, and evaluating the strength of body dynamic pain felt by the subject animal based on the evaluation result of the walking state. A method for evaluating body pain in bipedal walking of non-human primates, characterized by
[2] The evaluation of the walking state is performed based on one or more selected from the group consisting of the position of the center of gravity during walking, how to attach the hind limb to the floor, how to lift the hind limb, and the extension state of the knee joint. The evaluation method of body movement pain during biped walking of the non-human primate according to the above [1].
[3] The non-human primate of [1], wherein the evaluation of the walking state is performed by scoring the position of the center of gravity during walking, how to attach the hind limb to the floor, how to lift the hind limb, and the extension state of the knee joint. Method for evaluating body ache during bipedal walking of animals.
[4] The method for evaluating body pain during bipedal walking of a non-human primate according to any of [1] to [3], wherein the test animal is a cynomolgus monkey or a rhesus monkey.
[5] The evaluation method of body pain during bipedal walking of the non-human primate according to any one of the above [1] to [4], wherein the test animal is a pain model animal.
[6] A method for evaluating the action of the test substance for relieving body pain, which is a non-human primate ingested with the test substance as the test animal, which is any one of the above [1] to [5] The method for evaluating body movement pain during bipedal walking of non-human primates, and based on the evaluation result obtained to evaluate the presence or absence and the action strength of the test substance for relieving body movement pain. The evaluation method of the effect | action which relieve | moderates the dynamic pain of a to-be-tested substance characterized by the above.
[7] The strength of the body dynamic pain felt by the subject animal is compared with that before taking the test substance, and the strength of the body dynamic pain felt by the subject animal is the test substance When it is lower than before intake, it is evaluated that the test substance has an action of alleviating body pain of the test substance, and the strength of the body dynamic pain felt by the test animal is the above The body movement of the test substance of the above [6], wherein it is evaluated that the test substance has no effect on relieving body pain of the test substance when the strength is equal to or higher than the strength before taking the test substance. How to evaluate the effect of relieving pain.
[8] A method of screening a substance having an action of alleviating body pain, which comprises using a non-human primate ingested with a test substance as a test animal. The method for evaluating body motion pain during bipedal walking of a non-human primate, wherein the intensity of body motion pain felt by the subject animal is lower than before intake of the test substance A method of screening a substance having an action for relieving body pain, comprising selecting a test substance consumed by a subject animal as a candidate substance for a substance having an action for relieving body pain.

According to the method for evaluating body movement pain during biped walking of a non-human primate according to the present invention, body movement pain during biped walking of a non-human primate can be stably and objectively evaluated.
In addition, according to the evaluation method, non-human primates can be used to objectively evaluate the presence or absence and strength of the action of the test substance for relieving body dynamic pain, and further action for alleviating body dynamic pain. It is also possible to perform screening of substances that have

It is the figure which showed the one aspect | mode of the walking aid used in the body pain evaluation method which concerns on this invention. 1 (A) is a schematic perspective view, and FIG. 1 (B) is a side view. In Example 1, it is the figure which showed the gait disorder score (average value +/- standard error) of a diclofenac group (G2 group) and a vehicle group (G1 group). In Example 1, the results of Weight Bearing test of the vehicle group on day 36 after MIA administration (FIG. 3 (A)), and the results of Knee pressure test (FIG. 3) before and at 30 minutes after administration of morphine. (B) and the result of a gait disorder test (FIG. 3 (C)). In Example 2, it is the figure which showed the gait disorder score (mean value +/- standard error) of a MMx group and a Siamese group. In Example 2, the results of Weight Bearing test of MMx group on day 42 from MMx (FIG. 5 (A)) and the results of Knee pressure test (FIG. 5 (FIG. 5 (FIG. 5)) before and at 30 minutes after administration of morphine. B)) and the result of a gait disorder test (FIG. 5 (C)).

  The method for evaluating body movement pain during bipedal walking of a non-human primate according to the present invention (hereinafter sometimes referred to as "the method for evaluating body dynamic pain according to the present invention") And the test animal is allowed to walk freely with only the hind limbs while being placed on the walking aid holding the posture during biped walking with the hind limbs, and the step of evaluating the walking condition and the evaluation result of the walking condition Evaluating the strength of the body dynamic pain felt by the subject animal. When the subject animal feels a body pain, compared to when healthy (when not feeling a body pain), the posture at the time of walking collapses and no burden is placed on the part where the body pain is felt There is a tendency for crawling movements to be observed, and the tendency is greater as the intensity of body pain is stronger. That is, since the walking state of the subject animal is affected by the strength of the body dynamic pain felt by the subject animal, the strength of the body dynamic pain felt by the subject animal can be evaluated based on the walking state.

  In the method for evaluating body pain according to the present invention, the walking state is observed in a state where the test animal is placed on a walking aid that holds a posture during biped walking by the hind limbs. In order to evaluate body movement pain during bipedal walking by the hindlimb, it is necessary for the subject animal to walk only with the hindlimb without putting the forelimb on the floor. In the method for evaluating body movement pain according to the present invention, a non-human primate that originally walks on the limb by using the walking aid that holds the posture during biped walking by the hind limb, in certain time for sure only in the hind limb I can walk.

  The walking aid used in the method for evaluating body pain according to the present invention moves the subject animal according to biped walking of the subject animal while maintaining the posture at the time of biped walking by the hind limb to the subject animal. It assists bipedal walking. Here, "holding the posture during biped walking by the hindlimb" means that the forelimb is not attached to the floor, and the posture in which only the hindlimb is attached to the floor is maintained. More specifically, the walking aid includes a holding member having a holding portion for holding the neck of the subject animal, and a posture at which the height position of the holding portion indicates that the subject animal walks the two legs. A support main body for supporting the holding member and a wheel provided at a lower part of the support main body so as to be at a height position of the neck in; The holding portion is a hole smaller than the head of the subject animal and larger than the neck in plan view. The test animal placed on the walking aid can not be placed on the floor because the height from the floor of the neck is kept at the height in the upright posture by the hind limbs, and biped walking by the hind limbs The state is maintained. Moreover, since the said walking aid is equipped with the wheel, in the state installed in the said walking aid, a to-be-tested animal can walk freely back and forth and right and left. In addition, as the said walking aid, it is preferable to be considered so as to reduce the stress given to a test animal as much as possible.

  One aspect of the walking aid used in the method for evaluating body pain according to the present invention is shown in FIG. The walking aid 1 has a holding member 10, a support body 20, and wheels 30, is used on the floor F, and assists biped walking by the hind limbs of the non-human primate subject animal A. Used to

  The holding member 10 has a holding portion 14 for holding the neck of the subject animal A. The holding portion 14 is a hole smaller than the head of the subject animal A and larger than the neck in plan view. The holding member 10 is attached to the neck of the subject animal A at the holding portion 14.

  The support body 20 supports the holding member 10 at a predetermined height position. Specifically, with respect to the support body 20, the height position of the holding portion 14 of the support body 20 is biped walking of the subject animal A with reference to the height position of the neck in the posture when the subject animal A walks two legs. The holding member 10 is supported so as to be at the height position of the neck in time. As a result, the subject animal A on which the holding member 10 is mounted can hold the posture when walking on two legs.

  The wheels 30 are provided at the lower part of the support main body 20, and are capable of moving the entire walking aid 1 as the subject animal A walks on two legs. In the figure, the walking aid 1 has four wheels 30. Of course, the number of wheels 30 is not limited to four as long as it does not prevent biped walking of the subject animal A.

  The walking aid 1 further includes a forelimb holding unit 40 provided with a hand rest that allows the subject animal A to be held by the forelimbs in a biped walking posture. In the figure, the forelimb holding portion 40 is provided on the front lower portion of the holding member 10, but may be provided on the support main body 20. Many of the test animals A naturally live on a tree life, and when the biped is walked in a state where the forelimb has nothing, it causes stress. Therefore, the stress of the subject animal A can be reduced by providing the hand table (forelimb holding part 40) at a position where the subject animal A can be held by the forelimb.

  In the method for evaluating body pain according to the present invention, the walking condition is evaluated by freely walking only with the hind limbs in a state where the test animal is installed on the walking aid. The walking state is evaluated in comparison with the walking state (walking state at the time of healthy) of the same individual as the subject animal or the same individual but not feeling physical pain. When the walking state of the subject animal is the same as or similar to the walking state in the normal state, it is evaluated that the walking state of the subject animal is normal. When the walking state of the subject animal is different from the walking state at the healthy time, that is, when walking in a mode not observed in the walking state at the healthy time, the walking state of the subject animal is abnormal Evaluate.

  The walking state of the subject animal is preferably evaluated using, as an index, a mode that appears during walking when humans and non-human primates feel physical pain. Specifically, it is preferable to evaluate based on one or more selected from the group consisting of the position of the center of gravity when walking, how to attach the hind limb to the floor, how to lift the hind limb, and the extension state of the knee joint. The result of evaluating each item may be used as the evaluation of the walking state of the subject animal as it is, but the walking state can be evaluated more accurately by comprehensively judging and evaluating the evaluation of a plurality of items. .

  For example, when body pain is felt in one of the hind limbs, the center of gravity during walking is biased to the side of the hind limb where no body pain is felt (healthy foot centroid). Therefore, the walking state is evaluated as normal when the center of gravity at the time of walking is not biased to either the left or the right, and the walking state is evaluated as abnormal when the center of gravity at the time of walking is biased to either the left or the right.

  In addition, in the hindlimbs that feel physical pain, there is a tendency to walk with only the toes on the floor so as to lighten the weight load, and the motion to lift the hindlimbs that feel physical pain is also frequent It is observed. Therefore, when the area of the sole that reaches the floor is about the same as in the normal state, or when the action to lift the hind limb is not observed, the walking state is evaluated as normal, and the frequency when only the toe is attached is high, and the hind limb is lifted When motion is frequently observed, the walking state is evaluated as abnormal.

  Furthermore, when pain is felt in the knee joint, the range of motion of the knee joint becomes narrow, and the flexion and extension can not be smooth. Therefore, if the range of motion of the knee joint is 100% and bending and extension of the joint is possible without problems, the walking state is evaluated as normal, and the range of motion of the knee joint is narrowed, or the knee joint Although the range of motion of is 100%, the walking state is evaluated as abnormal if a force is applied to the foot at the time of extension and flexion of the joint or movement to endure pain is observed.

  The walking condition of the subject animal can be evaluated more objectively and semi-quantitatively by scoring the respective items and adding up the scores of all the items. For example, as shown in Table 1, the position of the center of gravity when walking, how to attach the hindlimb to the floor, how to lift the hindlimb, and the extension state of the knee joint include those shown in Table 1. Each of the four items (parameters) is scored, and the walking condition of the subject animal is evaluated based on the combined score of all the scores. When the score is 0, the walking state is evaluated as being normal, and as the score is increased, the walking state is evaluated as being worse (the degree of abnormality is larger).

  If the walking state is judged to be normal, the subject animal does not feel any body dynamic pain, or the strength of the body dynamic pain felt by the subject animal is evaluated to be extremely small. If the walking condition is judged to be abnormal, the subject animal is evaluated as feeling physical pain. When the walking state is evaluated by the score, the smaller the walking state score, the smaller the strength of the body dynamic pain felt by the subject animal, and the larger the walking state score, the larger the degree of abnormality of the walking state. It is evaluated that the intensity of the body dynamic pain felt by the subject animal is strong. It is good also as a score of the intensity | strength of the body dynamic pain which the test animal is feeling as it is, which evaluated the walking state.

  Non-human primates are classified into protozoans and protozoans. Protozoa include lemurs, loris, galagos, and tarsal monkeys, and true monkeys include spider monkeys, capuchin monkeys, marmosets, long-tailed monkeys, colobus and apes. The non-human primates to be tested in the method for evaluating body pain according to the present invention are preferably monkeys because the mechanism of pain and the response to analgesics are closer to human beings; It is more preferable that it is a colobus or apes, and it is more preferable that a cynomolgus monkey, a rhesus monkey, a Japanese monkey, a white gibbous monkey, a gorilla, an orangutan, a chimpanzee or a bonobo, and even more preferably a cynomolgus monkey or a rhesus monkey.

  As a test animal used in the method for evaluating body pain according to the present invention, a pain model animal is preferable, and OA induced by administration of MIA (Monosodium iodoacetate) to a joint or medial knee meniscectomy (Medial meniscectomy; MMx) Model animals and the like are more preferable. These pain model animals can be produced by a conventional method.

  The method for evaluating body pain according to the present invention can objectively evaluate body pain felt by non-human primates, so that the method evaluates the action relieving body pain of a test substance. It can be used for screening of substances having an action of alleviating body pain.

  Specifically, the non-human primate ingested with the test substance is used as the test animal, and the body dynamic pain evaluation method according to the present invention described above is performed, and the body of the test substance is obtained based on the obtained evaluation result. The presence or absence and the action strength of alleviating pain can be evaluated. For example, the strength of the body dynamic pain felt by the subject animal is compared with the strength of the body dynamic pain felt by the subject animal before taking the subject substance. The test substance has an effect of alleviating the body pain of the test substance when the strength of the body dynamic pain felt by the test animal is lower than before the test substance is ingested. Then, it is evaluated that the test substance has no effect on relieving body pain of the test substance when the strength is equal to or higher than the strength before intake of the test substance. In the method for evaluating body pain according to the present invention, the walking state of the subject animal is evaluated by a score, and the score is evaluated as it is as the score of the intensity of body pain, and the walking state of the subject animal When the score is lower than that before intake of the test substance, it can be evaluated that the action of alleviating body dynamic pain of the test substance is stronger as the reduction width of the numerical value of the score is larger.

  Further, the method for evaluating body pain according to the present invention described above is performed on a non-human primate ingested a test substance as a test animal, and the strength of body pain that the subject animal feels is determined by the subject. The test substance can be selected as a candidate substance of a substance having an action of alleviating body pain when the level is lower than before intake of the test substance. According to the method for assessing body dynamic pain according to the present invention, the intensity of the action of all test substances for relieving body pain can be evaluated as an objective numerical value (score), so it has the action of relieving body dynamic pain. By using the method for screening of substances, it is possible to efficiently select from among a plurality of test substances a substance having an action of alleviating body pain of desired intensity. The screening is performed on a plurality of test substances, but one test subject may be used to examine the strength of the action for alleviating body pain, and one test subject may An animal test may be used to examine the strength of the action to alleviate body pain of a plurality of test substances.

  EXAMPLES The present invention will next be described in more detail by way of examples, which should not be construed as limiting the invention thereto.

  In addition, animal experiments conducted in the following examples and breeding of used animals, etc. are the "Act on Protection and Management of Animals" (Law No. 105 of 1974) in Japan, "rearing of experimental animals and "Standards on storage and alleviation of pain" (2006 Ministry of the Environment notification No. 88), "Basic guidelines on implementation of animal experiments etc. at implementing agencies under the jurisdiction of Ministry of Health, Labor and Welfare" (notice of Ministry of Health, Labor and Welfare 2006), and "Animal It was conducted in compliance with the guidelines for appropriate implementation of experiments (designed by the Japan Science Council in 2006).

Example 1
The MIA-induced OA model monkey was used as a test animal, and the intensity of somatic pain during bipedal walking with and without the administration of the non-steroidal anti-inflammatory drug Diclofenac was evaluated.

<MIA induction OA model monkey>
MIA-induced OA model monkey is MIA (Sigma Aldrich; 1.5 mg of a 60 mg / mL solution) in the left knee joint cavity of cynomolgus monkeys of 7 to 10 years under anesthesia with 25 mg ketamine (Daiichi Sankyo Co., Ltd.) Made by injecting

<Diclofenac administration>
Diclofenac sodium salt (Sigma Aldrich) was orally administered once a day to five MIA-induced OA model monkeys such that the dose per monkey body weight was 1 mg / kg (diclofenac group: G2 group). Administration was performed for 35 days from the day after MIA administration.
As a control, 0.5 weight / volume% methylcellulose aqueous solution (manufactured by Wako Pure Chemical Industries, Ltd.) is orally administered once a day to five MIA-induced OA model monkeys in the same amount as the diclofenac administration group. Administered (vehicle group: group G1). Administration was performed for 35 days from the day after MIA administration.

<Walk disorder test>
Gait impairment tests were performed on days 0, 7, 14, 21, 28, and 35 after MIA injection on MIA-induced OA model monkeys in both the diclofenac group and the vehicle group. The measurements were performed under blinds, so that the gait disorder tester did not know which monkey was administered what. Also, on Day 0, a gait failure test was performed before MIA injection.
In the gait disorder test, first, the monkey was held in the gait aid shown in FIG. 1, and the gait was photographed with a video camera (manufactured by Panasonic Corporation) for about 1 to 3 minutes. The photographed images were analyzed, and the walking condition of monkeys was evaluated for each item according to the walking score table described in Table 1, and the total score of the scores of all items was used as the walking disability score of each monkey.

  The measurement results (mean value ± standard error) of gait disturbance scores of each group before MIA injection (day 0) and on days 1 to 35 from MIA injection are shown in FIG. As a result, in both groups, the gait disorder score was 0 and the gait status was normal before MIA injection (day 0), but the gait disorder score became high after day 7 after MIA injection An abnormality was observed in the walking state. On the 14th day after MIA injection, in the diclofenac group, the gait disorder score was significantly smaller than in the vehicle group (P <0.05, (t test)), and the degree of abnormality in the gait state was smaller. This is because diclofenac relieves pain caused by inflammation of the knee joint. From these results, the walking state in the state of being installed in the walking aid as shown in FIG. 1 reflects the intensity of the body dynamic pain felt by the non-human primate, and from the walking state, the body dynamic pain It is clear that the strength of the can be evaluated.

<Intensity change of pain by morphine administration>
For the MIA-induced OA model monkeys, changes in pain intensity due to administration of morphine were examined. Morphine was injected intramuscularly at a dose of 6 mg / kg per weight of monkey.
Specifically, for the vehicle group (5 animals) on the 36th day after MIA injection, the Weight Bearing test and Knee pressure on the left hind leg injected with MIA before and at 60 minutes after administration of morphine Tests were conducted, and further the gait disorder test was conducted as described above.

  Specifically, the Weight Bearing test was performed as follows. First, the monkey was held in a walking aid, and the hind limbs were placed on separate weighing scales (manufactured by Tanita Co., Ltd.). After confirming that the monkey was at rest and in stable position, measurements were started. The measurement was performed three times at each measurement time point. When the animal moved during the measurement, the value was excluded and the measurement was performed again, and the average of three times was taken as the measurement value at the measurement point. The load ratio (%) to the treatment side hind limb (left hind limb) was calculated by the following formula (1).

  The knee pressure test was performed as follows. First, the monkey was held in a walking aid, and the joint space in the knee and the medial femoral condyle were gently pushed for about 3 seconds using a pressure gauge (manufactured by Matsumiya Medical Seiki Seisakusho Co., Ltd.). The pressure value (kg) was determined as the measurement value at each measurement time point (cut-off value: 3 kg) when pain was recognized from the expression of the monkey or when withdrawal behavior such as withdrawal of the foot was observed. ). The measurement was performed three times at each measurement time point. The pain threshold (%) of the treatment side hind limb (left hind limb) was calculated by the following formula (2).

  The results of the Weight Bearing test are shown in FIG. 3 (A), the results of the Knee pressure test are shown in FIG. 3 (B), and the results of the gait disorder test are shown in FIG. 3 (C). ). As a result, compared to before morphine administration, the weight bearing on the left hind limb increases significantly (P <0.01 (correspondence t test)) significantly after 60 minutes from morphine administration, and the left knee joint bears Threshold increased, and the gait disability score decreased. This is because morphine relieves pain in the knee joint. Also from these results, the walking state in the state of being installed in the walking aid as shown in FIG. 1 reflects the intensity of the body dynamic pain felt by non-human primates, and It is clear that the intensity of the pain can be assessed.

Example 2
The MMx-induced OA model monkey was used as a test animal, and the intensity of body pain during bipedal walking was evaluated.

<MMx induction OA model monkey>
MMx-induced OA model monkeys under the anesthesia of 25 mg ketamine (manufactured by Daiichi Sankyo Co., Ltd.) and 25 mg somnopentyl (manufactured by Kyoritsu Pharmaceutical Co., Ltd.) under anesthesia, medial meniscectomy for the right knee joint of six 7 to 10 age cynomolgus monkeys Created by performing surgery (MMx group).
As a control, the right knee joint of three 7 to 10-year-old cynomolgus monkeys was prepared by performing a sham operation (incision of the skin at the same site as the operation performed on the MMx group) (sham group).

<Walk disorder test>
As in Example 1 before surgery (day 0) and on days 7, 14, 17, 21, 24, 28, 35, 42, 49 and 56 after surgery for both the MMx group and the sham group I did a gait disorder test. On Day 0, a gait disorder test was performed before surgery.

<Training>
On the 10th to 45th days of surgery, training was given to jump from one cage of the two cages placed at an interval of 2m to the other cage for 10 days to 45 days for both the MMx group and the sham group The Between the jumps, a break of 1 to 2 minutes was taken and 50 jumps were made per day. At the time of training and after the training, side dishes such as bananas and apples were given to prevent stress on monkeys.

  The measurement result (mean value +/- standard error) of the gait disorder score of each group in 0-56 days after surgery is shown in FIG. As a result, in both groups, the gait disorder score was 0 and the gait state was normal before the operation (day 0). In addition, in the sham group, the gait disorder score was almost 0 at 7 to 56 days after the operation, and the gait was normal. On the other hand, in the MMx group, the gait disorder score was 0 until the 14th day after the operation, but the gait disorder score became high after the 17th day after the operation, and an abnormality was observed in the gait state. In particular, on the 10th to 45th days after the surgery in which the training was performed, the gait disorder score increased over time, and on the 49th to 56th days after the surgery after the training, the gait disorder score decreased slightly Was. From the change in gait score, the left knee was stressed by training, and MMx-induced OA model monkeys felt pain, and the pain felt by monkeys became more severe with the continuation of training, but It became clear that the pain improved a little by the end.

<Intensity change of pain by morphine administration>
For the MMx-induced OA model monkeys, changes in pain intensity with morphine administration were examined. Morphine was injected intramuscularly at a dose of 6 mg / kg per weight of monkey.
Specifically, the Weight Bearing test and Knee pressure test were performed before and 60 minutes after the administration of morphine to the MMx group (six animals) on the 42nd day after surgery, and the gait disorder as described above I did a test. Each test was performed in the same manner as Example 1.

  The results of the Weight Bearing test are shown in FIG. 5 (A), the results of the Knee pressure test are shown in FIG. 5 (B), and the results of the gait disorder test are shown in FIG. 5 (C). ). As a result, compared to before morphine administration, 60 minutes after morphine administration significantly (Weight Bearing test P <0.05, Knee pressure test and gait disorder test P <0.01 (match t test) 2.) The weight load on the right hind leg (treated side) increased, the threshold of pressure that the right knee joint can withstand increased, and the gait disorder score decreased. From these results, it was confirmed that morphine relieved pain in the right knee joint.

  DESCRIPTION OF SYMBOLS 1 ... walking aid, A ... test | inspection animal, 10 ... holding member, 14 ... holding part, 20 ... support main body, 30 ... wheel, 40 ... forelimb holding part, F ... floor surface

Claims (8)

A method for evaluating body pain in a test animal, wherein the test animal is a non-human primate,
Evaluating the walking state by allowing the test animal to freely walk only with the hind limbs while being placed on a walking aid that holds the posture during biped walking with the hind limbs;
Evaluating the intensity of body dynamic pain felt by the subject animal based on the evaluation result of the walking state;
A method for evaluating body pain in bipedal walking of non-human primates, comprising:   The evaluation of the walking state is performed based on one or more selected from the group consisting of the position of the center of gravity during walking, how to attach the hind limb to the floor, how to lift the hind limb, and the extension state of the knee joint. The evaluation method of the body movement pain at the time of biped walking of the nonhuman primate as described in 1.   The non-human primate according to claim 1, wherein the evaluation of the walking state is performed by scoring the position of the center of gravity during walking, how to attach the hind limb to the floor, how to lift the hind limb, and the extension state of the knee joint. Evaluation method of body ache during bipedal walking.   The method for evaluating body pain during biped walking of a non-human primate according to any one of claims 1 to 3, wherein the test animal is a cynomolgus monkey or a rhesus monkey.   The evaluation method of body pain during biped walking of a non-human primate according to any one of claims 1 to 4, wherein the test animal is a pain model animal. A method for evaluating the action of a test substance to alleviate body pain, comprising:
A method for evaluating body pain during bipedal walking of a non-human primate according to any one of claims 1 to 5, wherein the non-human primate ingested the test substance is a test animal. An evaluation method of the action for alleviating body dynamic pain of a test substance, characterized by evaluating the presence or absence and the action strength of the test substance for alleviating the body dynamic pain based on the evaluated result. Comparing the intensity of body dynamic pain felt by the subject animal with that before taking the subject substance;
The test substance has an effect of alleviating the body dynamic pain of the test substance when the intensity of the body dynamic pain felt by the test animal is lower than before the intake of the test substance. To evaluate
The test substance has the effect of alleviating the body dynamic pain of the test substance when the strength of the body dynamic pain felt by the test animal is equal to or higher than the strength before taking the test substance. The evaluation method of the effect | action which relieve | moderates the body dynamic pain of the to-be-tested substance of Claim 6 to evaluate. A method of screening a substance having an action of alleviating body pain, comprising:
A method for evaluating body pain during bipedal walking of a non-human primate according to any one of claims 1 to 5, wherein the non-human primate ingested the test substance is a test animal,
When the intensity of the body dynamic pain felt by the subject animal is lower than that before taking the subject substance, the subject substance ingested by the subject animal has an effect of alleviating the body dynamic pain. A screening method of a substance having an action of alleviating body dynamic pain, characterized by selecting it as a candidate substance of a substance to have.

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