JP4340339B2 - Blood coagulation factor or blood coagulation control factor deficient model animal and method for producing the model animal - Google Patents

Description

【００２５】
【表３】

【００２６】
【表４】

【００２７】
上記事実は、外因系凝固因子であるサルＦＶＩＩがヒトＦＶＩＩａの投与により誘導された抗体によってその機能が中和されたか、もしくは、体内で産生された抗ヒトＦＶＩＩａ抗体と免疫複合体が形成され網内系により処理され代謝されたものと推察される。また、Ｎｏ.３サルの血漿中に正常サル血漿のＦＶＩＩ活性の中和活性が認められたことから、産生された抗ヒトＦＶＩＩａ抗体はサルＦＶＩＩ活性の中和能を有していることを強く示唆している。以上の結果は、サルにヒトＦＶＩＩａの静脈内投与を頻回行うことによりＦＶＩＩ欠損モデルが作出可能であることを示している。
【００２８】
実施例３
(ＦＩＸ欠損モデルのウサギでの考察)
(１)実施方法
ヒトＦＩＸをウサギに免疫し精製した抗ヒトＦＩＸウサギ抗体(ノルデディク社より購入)が正常ウサギ血漿中のＦＩＸへの中和活性を有しているかどうかを調べた。試験は、ウサギ血漿３００μｌに抗ヒトＦＩＸウサギ抗体を終濃度が０.０〜１.０ｍｇ/ｍｌになるように添加し３７℃で２時間インキュベ−トした後、残存するＦＩＸ活性を測定することによって実施した。
【００２９】
(２)試験結果及び考察
残存ＦＩＸ活性測定の結果、上記抗体濃度０.０〜１.０ｍｇ/ｍｌ以下ではウサギＦＩＸ活性の低下は確認されなかった(図６)。本結果は、ヒトＦＩＸを免疫原としてウサギで産生された抗体はウサギＦＩＸとは反応しないか、もしくはその中和活性の程度が低いことを示唆している。
【００３０】
実施例４
(ＦＩＸ欠損モデルのヤギでの考察)
(１)実施方法
ヒトＦＩＸをヤギに免疫し精製した抗ヒトＦＩＸヤギ抗体(ＣＥＤＡＲＬＡＮＥ社より購入)が正常ヤギ血漿中のＦＩＸへの中和活性を有しているかどうかを調べた。試験は、正常ヤギ血漿３００μｌに抗ヒトＦＩＸヤギ抗体を終濃度が０.０〜１.０ｍｇ/ｍｌになるように添加し３７℃で２時間インキュベ−トしたのち、残存するＦＩＸ活性を測定することによって実施した。
【００３１】
(２)試験結果及び考察
残存ＦＩＸ活性測定の結果、上記抗体濃度０.０〜１.０ｍｇ/ｍｌ以下ではヤギＦＩＸ活性の低下は確認されなかった(図７)。本結果は、ヒトＦＩＸを免疫原としてヤギで産生された抗体はヤギＦＩＸとは反応しないか、もしくはその程度が低いことを示唆している。
【００３２】
実施例３及び４の結果から、系統発生上サルより低い動物種ではヒト血液凝固因子を抗原として投与しても自己の凝固因子を中和する抗体が産生される可能性は極めて低いことが推定された。
【図面の簡単な説明】
【図１】ヒトＦＩＸをサルに投与したときのＰＴの変動を示す図である。
【図２】ヒトＦＩＸをサルに投与したときのＡＰＰＴの変動を示す図である。
【図３】ヒトＦＩＸをサルに投与したときのＦＩＸ力価の変動を示す図である。
【図４】ヒトＦＩＸをサルに投与したときの抗ヒトＦＩＸ抗体価の変動を示す図である。
【図５】ヒトＦＶＩＩａをサルに投与したときの抗ヒトＦＶＩＩａ/ＦＶＩＩ抗体価の変動を示す図である。
【図６】抗ヒトＦＩＸウサギ抗体によるウサギＦＩＸの中和試験における残存ＦＩＸ活性測定の結果を示す図である。
【図７】抗ヒトＦＩＸヤギ抗体によるヤギＦＩＸの中和試験における残存ＦＩＸ活性測定の結果を示す図である。[0001]
[Industrial application fields]
The present invention relates to a blood coagulation factor or blood coagulation control factor deficient model animal. Specifically, the present invention relates to a blood coagulation factor or blood coagulation control factor deficient model using a gagged animal and a method for producing the model animal. More specifically, a substance having the same function as a human-derived factor in a gagged animal body by immunizing a gagged animal with various methods using a human-derived blood coagulation factor or blood coagulation control factor as an immunogen The present invention provides a blood coagulation factor or blood coagulation control factor-deficient model produced by neutralizing and dysfunction and a method for producing the model animal.
[0002]
[Background Art and Problems to be Solved by the Invention]
A deficiency in blood clotting factor or its regulatory factor unbalances the normal blood clotting reaction and its control response, resulting in excessive bleeding and spontaneous thrombus. In particular, hereditary blood coagulation factor deficiency diseases include blood coagulation factor VIII (hereinafter sometimes referred to as FVIII) deficiency, hemophilia A, blood coagulation factor IX (hereinafter sometimes referred to as FIX) deficiency Have hemophilia B. These diseases are characterized by clinically severe recurrent deep organ bleeding (joint, muscle, central nervous system, gastrointestinal bleeding, etc.), and there are about 3000 patients in Japan.
[0003]
Hemophilia patients are always at risk of bleeding, and in order to control their bleeding, that is, to prevent bleeding and stop bleeding, FVIII or FIX expressed in various cells is extracted and purified from pooled plasma or expressed by genetic recombination. Replacement therapy is administered. However, since such a treatment method is originally performed on patients who are deficient in FVIII or FIX, neutralizing antibodies may be produced in some patients considering these administered blood coagulation factors as foreign substances. It is said that approximately 6.0% of hemophilia patients have hemophilia inhibitors in which replacement therapy is ineffective in this way, which is one of the most serious side effects associated with replacement therapy. (Michio Matsuda, Koji Suzuki, Hemostasis / Thrombus / Fibrinolysis, published by Chugai Medical, 1994, Tadashi Kamiya, Recent Topics on p.472-479).
[0004]
For the treatment of the above-mentioned patients with hemophilia inhibitors, (1) an excessive amount of blood coagulation factor VIII and blood coagulation factor IX is administered to surpass the produced antibody. (2) Bypass preparations (Proplex (trade name), Autoplex (trade name), Fiber (trade name) that do not go through FVIII or FIX in the blood coagulation pathway, that is, cause blood coagulation by the sideway (bypass) route) ) Is administered. However, overdose of deficient factors has quantitative and economic limitations. Bypass preparations have a strong ability to produce thrombin, which is the final stage of blood coagulation. (N. Eng. J. Med. Vol. 303, p. 421-425, 1980), and development of a new preparation satisfying efficacy and safety was awaited. It is.
[0005]
A drug efficacy model using laboratory animals is indispensable for formulation development. So far, the efficacy evaluation of various bypass preparations has been used in dog models (hemophilia dogs) with congenital FVIII or FIX deficiency and transient hemophilia inhibitor models by administration of anti-FVIII antibody and inhibitor patient plasma to various animals. (Proc. Natl. Acad. Sci., Vol. 86, p. 1382-1386, 1989; Thrombosis and Haemostasis, vol. 77, p. 591-599, 1997). In recent years, transgenic mice have been used to produce blood coagulation factor VIII and blood coagulation factor IX deficient mice (Nature Genetics, vol. 10, p. 119-121, 1995; Blood, vol. .88, p. 3446-3450, 1996; Blood, vol. 91, p. 784-790, 1998). However, because these animal models are separated from humans, it is possible to obtain test results that are sufficiently applicable to humans in terms of the pathology of inhibitors, blood kinetics of administered drugs, and evaluation of drug efficacy. This is one of the reasons for the development of new therapeutic agents for hemophilia inhibitors.
[0006]
[Means for Solving Problems, Structure of the Invention]
As a result of intensive efforts to solve the above problems, the inventors of the present application have obtained a model animal using a gagged animal that suitably reproduces a human blood coagulation factor or blood coagulation factor deficient state, and the model animal. The present invention has been completed which provides a simple production method.
The present invention is outlined below.
[0007]
A human blood coagulation factor or blood coagulation control factor-deficient model animal using the gagged animal of the present invention was produced in an animal body by immunizing a monkey animal with a human-derived blood coagulation factor or blood coagulation control factor. This is an inhibitor-bearing patient model that suitably reproduces a state in which an antibody against a human immunogen neutralizes a blood coagulation factor or blood coagulation factor in an animal body. In addition, the present invention also provides a method by which the human blood coagulation factor or blood coagulation control factor deficient model animal can be easily produced.
[0008]
The human-derived blood coagulation factors used in the present invention include blood coagulation factor XII, blood coagulation factor XIII, blood coagulation factor XI, blood coagulation factor X, blood coagulation factor IX, blood coagulation factor VIII, blood There are coagulation factor VII, blood coagulation factor V, prothrombin and fibrinogen. In addition, blood coagulation regulators include a series of serine protease inhibitors such as protein C, protein S, protein Z, thrombomodulin, TFPI (Tissue Factor Pathway Inhibitor), AT-III (Antithrombin-III) and the like.
The factor used as an immunogen in the present invention includes blood, cell-derived factors as well as those produced by a chemically synthesized peptide based on the amino acid sequence or a gene recombination technique as long as it is derived from human. Further, the gagged animals as model animals of the present invention are preferably phylogenetically large monkeys greater than true monkeys, such as cynomolgus monkeys, and these are preferably used. It is not particularly limited as long as it exhibits a control factor deficient state.
[0009]
The immunization method is not limited as long as it is a method capable of producing a neutralizing antibody. Desirably, immunization by inoculating the antigen subcutaneously, intradermally or intramuscularly is recommended, and according to these immunizations administered The original amount can be lower than the intravenous dose. In inoculation with an antigen, it is effective to use an adjuvant having an immunostimulatory effect, but the use of Freund's complete adjuvant and incomplete adjuvant in combination gives favorable results. Suitable antigen and adjuvant amounts and dosing schedules are as follows.
A general animal immunization method of heterologous protein is applied, and several mg / kg of antigen is mixed with Freund's complete adjuvant and administered to a plurality of subcutaneous sites. Three to four weeks later, the antigen is similarly mixed with Freund's incomplete adjuvant and administered subcutaneously to multiple sites.
[0010]
Hematology indicators of disease to be modeled after initiation of immunization, for example (1) tissue thromboplastin time and activated partial thromboplastin time, (2) coagulation factor activity, (3) anti-human FVIIa or anti-human FIX antibody, (4) neutralization test, and (5) coagulation factor or blood coagulation control factor deficiency targeting experimental animals if measurement of bleeding time and blood volume over time and observation of monkeys at the same time It can be confirmed that it was able to be guided to the state. After the induction is completed, administration of the developed drug enables evaluation of the drug efficacy.
[0011]
The following outlines the hematology indicators of the disease and the measurement principles of the indicators.
(1) Tissue thromboplastin time (PT) and activated partial thromboplastin time (APTT)
The hemostatic reaction of blood is classified into a primary hemostatic reaction based on adhesion / aggregation of platelets to a vascular injury site and a secondary hemostatic reaction based on subsequent fibrin formation. There are two pathways in the coagulation reaction: an extrinsic coagulation reaction and an intrinsic coagulation reaction. PT is the clotting time when tissue thromboplastin / phospholipid / Ca ²⁺ is added to plasma, and APTT is the clotting time when actin / phospholipid / Ca ²⁺ is added to plasma. PT mainly detects abnormalities in extrinsic blood coagulation factor VII (hereinafter sometimes referred to as FVII), and APTT mainly detects abnormalities in intrinsic coagulation factors (FXII, FXI, FIX, FVIII).
[0012]
(2) Coagulation factor activity measurement method Coagulation factor activity is measured using plasma deficient in each coagulation factor. For example, when measuring the activity of FVII, a sample is added to FVII-deficient plasma, and coagulation is initiated by a mixed solution of tissue thromboplastin / phospholipid / Ca ²⁺ , and the activity (fold / ml). Similarly, in the case of FIX, a sample is added to FIX-deficient plasma, and after adding actin / phospholipid mixed solution, Ca ²⁺ is further added to initiate a coagulation reaction, and the activity when standard plasma is made 1-fold ( Times / ml).
[0013]
(3) Measurement of anti-human FVIIa or anti-human FIX antibody The increase in IgG antibody produced in a model animal is obtained by immobilizing human FVIIa or human FIX on a solid phase plate, developing the sample, and then antiperoxidase-conjugated anti-antibody. After reacting with the IgG antibody, confirmation is made by measuring the color intensity of the hydrolyzed chromogenic substrate.
[0014]
(4) Neutralization test The degree of whether the antibody produced in the model animal has the ability to neutralize FVII and FIX in the plasma is determined by mixing the sample diluted in the model animal standard plasma 1: 1. The dilution factor at which the activity is neutralized by 50% is evaluated as Bethesda units (BU).
[0015]
(5) Measurement of bleeding time and amount of bleeding A cut is made with a thin plate (trade name) (Organon Technica) on the tail of a model animal, and blood is put on Whatman 3MM (trade name) filter paper (Whatman Corp.) every 30 seconds. The time when adsorption is stopped and adsorption is stopped is defined as bleeding time. After the primary bleeding is completed, secondary bleeding causes rebleeding by wiping the cut site with absorbent cotton three times, and this is designated as secondary bleeding. The amount of bleeding is calculated by extracting the adsorbed blood with an ammonia solution and quantifying the amount of hemoglobin.
[0016]
【The invention's effect】
The present invention provides a human blood coagulation factor or blood coagulation control factor-deficient model animal that could not be easily reproduced conventionally, and a method for producing the model animal. As a result, barriers to development of therapeutic agents for hemophilia inhibitors are reduced, and further progress in the development of such therapeutic agents is expected.
EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.
[0017]
【Example】
Example 1
(FIX deficiency model using gagged animals)
(1) Implementation method 60 liters of human fresh frozen plasma was cooled and thawed, and the supernatant obtained by centrifuging the precipitate fraction was added to an anion exchanger (DEAE-Sephadex A-40, Pharmacia). It was washed thoroughly with 20 mM citric acid / 0.1 M NaCl (pH 7.0), and the vitamin K-dependent coagulation factor fraction having the Gla domain was eluted with 20 mM citric acid / 0.5 M NaCl (pH 7.0). . 300 liters of eluate was developed on an anti-FIX monoclonal antibody-immobilized affinity gel pre-equilibrated with 50 mM Tris / 150 mM NaCl / 5.0 mM CaCl ₂ (pH 8.0), and 50 mM Tris / 2.5 M NaCl / 5.0 mM CaCl ₂ After washing with (pH 8.0) buffer, further washing with 50 mM Tris / 30 mM NaCl / 5.0 mM CaCl ₂ (pH 8.0) and elution with 50 mM Tris / 30 mM NaCl / 10 mM EDTA · 2Na (pH 7.4) FIX fraction was obtained. The obtained FIX fraction was spread on a virus removal membrane (Bemberg microporous membrane, Asahi Kasei) that had been packed in advance with a buffer of 50 mM Tris / 30 mM NaCl (pH 8.0) to obtain a filtrate. 40 mg of human FIX having a purity of 90% or more was obtained in the filtrate.
[0018]
Human FIX was mixed with 3 cynomolgus monkeys (No. 1 male-6 years old, No. 2 male-6 years old, No. 3 female-9 years old) with human FIX and Freund's complete adjuvant in a 1: 1 ratio. As an initial immunization to 5 mg / head, human FIX and Freund's incomplete adjuvant were administered subcutaneously in the same manner on the 28th day after the first immunization for further immunization. As a monkey coagulation test, PT, APTT, FDIX activity, and FIX antibody titer were measured. In addition, No. 1 monkey was administered 45 μg / kg of the FVIIa preparation which is one of the bypass preparations. In the administration test of the FVIIa preparation, bleeding time and bleeding amount were measured before administration, 30 minutes after administration, and 360 minutes after administration.
[0019]
(2) Results In all three cynomolgus monkeys, PT did not change compared to before the start of immunization (Fig. 1). Fig. 4) was observed. The FIX activity decreased as the antibody titer increased (FIGS. 3 and 4), and the neutralization activity of No. 1 monkey 35 days after the start of immunization was 12 BU. This phenomenon was considered to be caused by the antibody induced by the human FIX antigen forming an immune complex with its own FIX. The No. 1 monkey out of the 3 animals maintained a FIX activity of less than 5% over 3 weeks after the boost (FIG. 3), during which transient hematuria and purpura were observed in the upper arm, Symptoms like disease B inhibitor. Further, FVIIa was administered at 45 μg / kg to No. 1 monkey 3 weeks after the booster immunization, and as a result, the bleeding time was shortened and the bleeding amount was reduced (Table 1). Therefore, it was confirmed that this model is effective as a medicinal efficacy evaluation system for bypass formulations.
[0020]
[Table 1]
──────────────────────────────────
Item Before administration 30 minutes after administration 360 minutes after administration Bleeding time Primary bleeding time (min, sec) 9.22 4.02 3.42
Secondary bleeding time (min, sec) 4.15 2.29 5.27
Bleeding volume Primary bleeding volume (μl) 93.1 56.6 35.5
Secondary bleeding volume (μl) 69.5 24.3 49.0
──────────────────────────────────
The test was performed twice and the average value was displayed.
Example 2
(FVII deficiency model using monkeys)
(1) Execution method 3000 liters of human fresh frozen plasma was cooled and thawed, and the supernatant obtained by centrifuging the precipitate fraction was added to an anion exchanger (DEAE-Sephadex A-40, Pharmacia). It was washed thoroughly with 20 mM citric acid / 0.1 M NaCl (pH 7.0), and the vitamin K-dependent protein fraction having the Gla domain was eluted with 20 mM citric acid / 0.5 M NaCl (pH 7.0). 300 liters of eluate was developed on an anti-FVII monoclonal antibody immobilized affinity gel pre-equilibrated with 50 mM Tris / 150 mM NaCl / 5.0 mM CaCl ₂ (pH 8.0), and 50 mM Tris / 2.5 M NaCl / 5.0 mM CaCl ₂ After washing with (pH 8.0) buffer, further washing with 50 mM Tris / 30 mM NaCl / 5.0 mM CaCl ₂ (pH 8.0) and elution with 50 mM Tris / 30 mM NaCl / 10 mM EDTA · 2Na (pH 7.4) FVII fraction was obtained. The obtained FVII fraction was spread on a virus removal membrane (Bemberg microporous membrane, Asahi Kasei) that had been packed in advance with a buffer of 50 mM Tris / 30 mM NaCl (pH 8.0) to obtain a filtrate. The FVII purity of the obtained filtrate was 85%. The above-described 50 ml of 0.2 mg / ml FVII solution was developed on a DEAE-Sepharose Fast Flow (Pharmacia) column of φ5.0 mm and height of 5.0 cm, which had been equilibrated with 50 mM Tris / 30 mM NaCl (pH 8.0) in advance. Upon activation, 800 mg of 95% pure FVIIa was obtained.
[0022]
The above human FVIIa was intravenously administered 3.3 mg / kg every day for 28 days to 3 4-year-old cynomolgus monkeys (No.1, No.2 were male and No.3 was female). After the administration, PT, APTT, FVII activity, and FVIIa antibody titer were measured as a coagulation test. Further, FVII neutralizing ability was measured using No. 3 monkey plasma on the 28th day after administration.
[0023]
(2) Results and discussion Human FVIIa was continuously administered to 3 monkeys daily for 28 days, and coagulation tests were conducted before and after 28 days. As a result, no change in APTT was observed in all 3 monkeys. , Significant extension of PT was observed (Table 2 (PT fluctuation), Table 3 (APTT fluctuation)). On the 14th day after the start of administration, a decrease in FVII activity was observed, and on the 28th day, it decreased to below the detection sensitivity (Table 4 (variation of FVII activity)). Further, the anti-human FVIIa / FVII antibody titer was confirmed to increase on the 14th day after the start of administration, and further increased on the 28th day (FIG. 5). The neutralizing activity of the No. 3 monkey plasma on the 28th day was measured and found to be 5 BU.
[0024]
[Table 2]

[0025]
[Table 3]

[0026]
[Table 4]

[0027]
The above facts indicate that monkey FVII, an exogenous coagulation factor, was neutralized by an antibody induced by administration of human FVIIa, or an immune complex was formed with an anti-human FVIIa antibody produced in the body. It is presumed that it was processed and metabolized by the internal system. Further, since the neutralizing activity of FVII activity of normal monkey plasma was observed in the plasma of No. 3 monkey, it was strongly confirmed that the produced anti-human FVIIa antibody has the ability to neutralize monkey FVII activity. Suggests. The above results indicate that an FVII deficient model can be created by repeatedly administering human FVIIa intravenously to monkeys.
[0028]
Example 3
(Consideration of FIX deficiency model in rabbit)
(1) Implementation method It was examined whether or not an anti-human FIX rabbit antibody (purchased from Nordedik) obtained by immunizing and purifying rabbits with human FIX has a neutralizing activity to FIX in normal rabbit plasma. In the test, anti-human FIX rabbit antibody was added to 300 μl of rabbit plasma to a final concentration of 0.0 to 1.0 mg / ml, incubated at 37 ° C. for 2 hours, and then the remaining FIX activity was measured. Carried out by.
[0029]
(2) Test results and discussion As a result of measurement of residual FIX activity, a decrease in rabbit FIX activity was not confirmed at the antibody concentration of 0.0 to 1.0 mg / ml or less (FIG. 6). This result suggests that antibodies produced in rabbits using human FIX as an immunogen do not react with rabbit FIX or have a low degree of neutralizing activity.
[0030]
Example 4
(Consideration of FIX deficiency model in goat)
(1) Implementation method It was examined whether or not an anti-human FIX goat antibody (purchased from CEDALANE) obtained by immunizing and purifying human FIX in goats has neutralizing activity on FIX in normal goat plasma. In the test, anti-human FIX goat antibody was added to 300 μl of normal goat plasma to a final concentration of 0.0 to 1.0 mg / ml, and incubated at 37 ° C. for 2 hours, and then the remaining FIX activity was measured. Was carried out.
[0031]
(2) Test results and discussion As a result of measurement of residual FIX activity, no decrease in goat FIX activity was confirmed at the antibody concentration of 0.0 to 1.0 mg / ml or less (FIG. 7). This result suggests that antibodies produced in goats with human FIX as an immunogen do not react with goat FIX or the degree thereof is low.
[0032]
From the results of Examples 3 and 4, it is estimated that in animal species that are phylogenetic lower than monkeys, it is very unlikely that antibodies that neutralize their own coagulation factors will be produced even when human blood coagulation factors are administered as antigens. It was done.
[Brief description of the drawings]
FIG. 1 is a graph showing changes in PT when human FIX is administered to monkeys.
FIG. 2 shows changes in APPT when human FIX is administered to monkeys.
FIG. 3 is a graph showing changes in FIX titer when human FIX is administered to monkeys.
FIG. 4 shows changes in anti-human FIX antibody titer when human FIX is administered to monkeys.
FIG. 5 shows changes in anti-human FVIIa / FVII antibody titer when human FVIIa is administered to monkeys.
FIG. 6 is a graph showing the results of measurement of residual FIX activity in a rabbit FIX neutralization test using an anti-human FIX rabbit antibody.
FIG. 7 is a graph showing the results of measurement of residual FIX activity in a goat FIX neutralization test using an anti-human FIX goat antibody.

Claims (10)

Human-derived blood coagulation factor or blood coagulation control factor is a vitamin K-dependent factor, and the human-derived factor is administered to a gagged animal to induce an immune reaction, which is the same as the human-derived factor in vivo in a gagged animal A blood coagulation factor or blood coagulation factor inhibitor patient model animal produced by neutralizing a functional blood coagulation factor or blood coagulation control factor by an immune reaction. The blood coagulation factor or blood coagulation control factor inhibitor patient model animal according to claim 1, wherein the monkey animal is a large monkey phylogenetically greater than or equal to a true monkey. Human-derived blood clotting factor or a blood coagulation control factors, blood coagulation factor X, blood clotting factor IX, blood clotting factor VII, prothrombin, is selected from protein C, in any one of claims 1 or claim 2 The blood coagulation factor or blood coagulation factor inhibitor patient model animal described. The blood coagulation factor or blood coagulation control factor inhibitor according to any one of claims 1 to 3 , wherein the human-derived blood coagulation factor or blood coagulation control factor is selected from blood coagulation factor IX and blood coagulation factor VII. Patient model animal. Human-derived blood coagulation factor or blood coagulation control factor is a vitamin K-dependent factor, and the human-derived factor is administered to a gagged animal to induce an immune response, and is identical to the human-derived factor in vivo in a gagged animal A method for producing a blood coagulation factor or blood coagulation factor inhibitor patient model animal produced by neutralizing a functional blood coagulation factor or blood coagulation control factor by an immune reaction. 6. The method for producing a blood coagulation factor or blood coagulation control factor inhibitor patient model animal according to claim 5, wherein the monkey animal is a phylogenetically large monkey that is equal to or greater than a true monkey. Human-derived blood clotting factor or a blood coagulation control factors, blood coagulation factor X, blood clotting factor IX, blood clotting factor VII, prothrombin, is selected from protein C, to claim 5 or claim 6 A method for producing the described inhibitor patient model animal. The method for producing an inhibitor patient model animal according to any one of claims 5 to 7, wherein the human-derived blood coagulation factor or blood coagulation control factor is selected from blood coagulation factor IX and blood coagulation factor VII. The method for producing an inhibitor patient model animal according to any one of claims 5 to 8 , wherein the human-derived blood coagulation factor or blood coagulation control factor is administered subcutaneously or intravenously. 10. The method of producing an inhibitor patient model animal according to claim 9 , wherein the human-derived blood coagulation factor or blood coagulation control factor is administered by using Freund's complete adjuvant or Freund's incomplete adjuvant as an immunostimulating agent during subcutaneous administration. Method.

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