JPH0429680B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to chemically modified derivatives of blood coagulation factors, methods for producing the same, and blood coagulation promoters containing these.

Hemophilia is one of the diseases in which blood coagulation activity is reduced. In hemophilia, there is a congenital deficiency or malfunction of coagulation factors, which causes symptoms such as large hematomas occurring spontaneously or due to minor trauma, or prolonged hemostasis, but treatment is not required. The only solution is to supplement the missing factors. However, even with this replacement therapy, the blood half-lives of the coagulation factor deficient in hemophilia A or Von-Wil-Brown disease and the coagulation factor deficient in hemophilia B are 15 hours and 25 hours, respectively. In order to maintain coagulation activity, administration through frequent injections has been required. Moreover, these coagulation factors are unstable and easily degraded in the gastrointestinal tract, so oral administration is not expected to be effective.
I had no choice but to choose intravenous injection or drip.
As a result, since treatment for this disease continues throughout the patient's life, the burden of treatment is heavy not only on the patient but also on the doctor and family, and this has been a hindrance to the patient's social life.

As a result of repeated studies to reduce the burden on patients with these blood coagulation factor deficiency or malfunctioning diseases, as well as their doctors and families, the present inventors have found that the duration of activity in the blood can be extended, and oral We have discovered a novel blood coagulation factor derivative that can be administered and completed the present invention.

Therefore, an object of the present invention is to provide a novel blood coagulation factor derivative in which a polyethylene glycol derivative is bonded to the amino acid side chain of a coagulation factor that maintains coagulation activity in blood.

Another object is to provide a method for producing the above-mentioned novel blood coagulation factor derivatives.

Still another object is to provide a blood coagulation promoter that contains the above novel blood coagulation factor derivative and can be administered orally.

In the amino acid side chain of the novel blood coagulation factor of the present invention,
General formula RO(CH ₂ CH ₂ O) _o -A- (wherein, R is a hydrogen atom, an alkyl group, or an alkanoyl group, n is an integer, and A is an active residue (coupling) capable of bonding an amino group. agent), and the molecular weight _of RO( _CH2CH2O ) _o is 200~
20000) are bonded together.

The blood coagulation factors referred to here are factors derived from human plasma and necessary to complete the blood coagulation cascade, and their active forms (a).
a), fragments, and complexes.

In addition, examples of alkyl groups that are substituents of polyethylene glycol include methyl, ethyl, and stearyl groups, and examples of alkanoyl groups include acetyl and propionyl groups.
A methyl group is particularly preferred. The molecular weight of polyethylene glycol is 1000 to 20000.
10000 is preferred.

The coupling agent is not particularly limited as long as it can bind polyethylene glycol and further bind to the amino group of the amino acid side chain of a blood coagulation factor. Reacting polyethylene glycol and chloroacetic acid ethyl ester, then treating with hydrazine,
Acyl azide obtained by treating the obtained hydrazide with nitrous acid, and p
- p-nitropenzyl ether of polyethylene glycol by reacting nitrobenzyl chloride, p-nitrophenyl glyceryl ether, etc., 3
Examples include diazonium salts obtained by obtaining -(p-nitrophenoxy)-2-hydroxypropyl ether, reducing the nitro group, and then diazotizing it, with cyanuric halides being particularly preferred. Also,
In the case of this cyanuric halide, after binding polyethylene glycol and blood coagulation factors, the remaining halogen atoms may be substituted with hydroxyl groups.

The novel blood coagulation factor derivative of the present invention comprises a blood coagulation factor and the general formula: RO(CH ₂ CH ₂ O) _o -A-X (wherein R is a hydrogen atom, an alkyl group or an alkanoyl group, and n is an integer) , A represents an active residue capable of binding an amino group, and the molecular weight of RO(CH ₂ CH ₂ O ₎ is 200 to 20,000). Obtained by reaction.
The reaction must be carried out under conditions that minimize the decrease in coagulation activity of blood coagulation factors. That is, it is preferable to carry out the reaction at a low temperature while controlling the pH using a buffer solution or the like.
The reaction time varies depending on the type of coupling agent, etc., but is preferably from several minutes to 24 hours. PH of buffer solution
can be arbitrarily selected as long as the blood coagulation factor is not completely deactivated, but is preferably in the range of 5 to 9, and is also determined depending on the type of coupling agent. The amount of the reagent for the polyethylene glycol and coupling agent conjugate is determined depending on the desired degree of modification. In other words, if you want to increase the degree of modification, increase the reagent concentration at a high pH,
If you try to reduce the degree of modification by conducting the reaction for a long time,
Reduce the reagent concentration and run the reaction for a short time at a low pH. By appropriately combining these modification conditions,
A novel blood coagulation factor derivative that is stable, long-acting, and can be administered orally is obtained. In addition, after the reaction of the conjugate of blood coagulation factors, polyethylene glycol, and a coupling agent is completed, unreacted reagents can be removed using biochemical methods known per se that do not reduce blood coagulation factor activity, such as gel filtration, dialysis, and ion exchange chromatography. chromatography, affinity chromatography, etc. alone or in combination. The produced new blood coagulation factor derivatives are stored in a state containing a buffer solution or salt, frozen as a single product, or lyophilized.

The activity of the novel blood coagulation factor derivative obtained as described above varies depending on the type of conjugate of polyethylene glycol and coupling agent used and the degree of modification. For example, when blood coagulation factor is modified with monomethyl ether polyethylene glycol-4,6-dichloro-1,3,5-triazine and the activity is measured by the correction effect of the coagulation time of factor-deficient plasma, a substance with a molecular weight of 550 has a molecular weight of 10mM. The modified factor showed 113.8% of the unmodified factor, and the factor with a molecular weight of 15,000 showed 66.6% at 2 mM. Since these also contain factor 1, when their activity was measured in the same way as factor 1 using factor-deficient plasma, the factor activity was 210%, 115.1%, and 115.1%, respectively.
Factor activity is 115.3, 106.1%, factor activity is
It showed 104.9, 83.5%. On the other hand, when polyethylene glycol is modified with a polyethylene glycol with no terminal substitution, the activity of factor modified at 5mM with a molecular weight of 6000 is 71.8%, and the activity of factor modified with 5mM with a molecular weight of 3350 is 77.5%. It was hot. Activated polyethylene glycol terminated in stearyl ether and having a molecular weight of 3200, i.e. monostearyl ether polyethylene glycol-4,6-dichloro-
The activity of factor modified with 1,3,5-triazine at 3mM was 74.0%, the factor activity was 157.3%,
Factor activity is 110.3%, factor activity is 89.4%
It was hot. The molecular weight is stearyl ester at the end.
The activity of factor modified with 2700 activated polyethylene glycol at 5mM was 98.1%, factor activity 139.1%, factor activity 108.9%, and factor activity 91.4%.

Next, the relationship between modification reagent concentration and activity will be shown. For example, monomethyl ether polyethylene glycol-4,6-dichloro-1,3,5 with an average molecular weight of 5000
- For triazine-modified factor, 91.2% of unmodified factor at 1.72mM, 97.9% at 2.44mM, and 97.9% at 4.2mM.
113% respectively. These factor activities are 104.4, 105.5, 203%, factor activity is 108.4, 113.3, 104.7%, factor activity is
The percentages were 87.4, 74.7, and 102.5%.

Furthermore, when the novel blood coagulation factor derivative modified with polyethylene glycol obtained in the present invention was orally administered to dogs, it was observed that it migrated into the blood. For example, the above-mentioned monomethyl ether polyethylene glycol-4,6-dichloro- with an average molecular weight of 5000
Modified with 1,3,5-triazine Reagent concentration 4.2mM
When the modified factor was mixed into milk and administered orally, its transfer into the blood was observed, and its physiological activity was sustained. That is, when blood is collected and citrated plasma is separated at 1, 2, 4, 6, and 8 hours after oral administration, the partial thromboplastin time (PTT) of these plasmas is lower than that of the plasma before administration.
and prothrombin time (PT) were both shortened, and the coagulation time correction effect of factor-deficient plasma was also significant. In addition, the activity of factor 1 was sustained, indicating that these coagulation factors coexisting in factor preparations were also modified with polyethylene glycol.
It is thought that it was absorbed from the gastrointestinal tract and transferred to the bloodstream. In addition, when unmodified factor was orally administered, the correction effect was within the range of physiological fluctuations. In addition, as a control experiment, a modification reagent, monomethyl ether polyethylene glycol-4,6-
Oral administration of dichloro-1,3,5-triazine alone was also performed and a comparative study was performed, but no significant change in PTT or PT was observed. The above results are the first
It is shown in Figs. In addition, in an oral administration experiment of modified factor prepared in the same way with a reagent concentration of 1.72mM, which is less than half of that in the previous experiment, the clotting time correction effect of deficient plasma was small, allowing absorption from the gastrointestinal tract after oral administration. It can be seen that a certain degree of modification is required in order to normalize.

Next, blood coagulation factor was added to monomethyl ether polyethylene glycol-4 with an average molecular weight of 5000,
2.1 m of 6-dichloro-1,3,5-triazine
When modified with M, 2.94mM and 4.2mM concentrations and orally administered as a mixture to dogs, transfer of factor into the blood was observed. That is, both PTT and PT of the plasma of treated dogs were shortened compared to before administration, and the coagulation time correction effect of factor-deficient plasma was also significant. Moreover, these effects were observed even 8 hours after administration. On the other hand, when unmodified factor was orally administered as a control experiment, a temporary coagulation time correction effect appeared after 2 hours, but disappeared after 4 hours. The above results are shown in FIGS. 6 to 8. From the above, it can be seen that the polyethylene glycol-modified blood coagulation factor derivative is an excellent blood coagulation promoter that can be administered orally and has a prolonged duration in blood.

When the novel blood coagulation factor derivative of the present invention is used as a medicine, the factor derivative is used for hemophilia A or Von Willebrand disease, the factor derivative is used for hemophilia B, and the factor derivative is used for hemophilia B. It is also used for debilitating or consumptive bleeding, and administration methods include intravenous injection, drip, and oral administration. Currently, commercially available blood coagulation factor preparations contain amino acids and salts for the purpose of adjusting pH and osmotic pressure, but the novel blood coagulation factor of the present invention may contain these or other known additives and excipients. There is no problem in adding stabilizers, etc. Further, as a dosage form for oral administration, additives and excipients can be added to form powders, granules, tablets, capsules, etc. In addition, it is more preferable to use an enteric-coated dosage form.

The dosage varies depending on the type of blood coagulation factor to be modified, and the activity of the blood coagulation factor derivative of the present invention tends to be slightly lower than that of the blood coagulation factor used. As it exhibits a markedly prolonged blood half-life,
The single dose is preferably equal to or lower than the dose of the blood coagulation factor used. For example, a preparation containing the derivative of the present invention modified with human blood coagulation factor may be administered by intravenous injection or intravenous drip infusion for 1 day.
It can be administered at a dose of 5 to 30 units/Kg (250 to 2000 units for adults) depending on the symptoms.

In the case of oral administration, higher doses are required because deactivation in the gastrointestinal tract is expected compared to parenteral administration such as intravenous injection.

Furthermore, since the blood coagulation factor derivative of the present invention has a prolonged activity duration, the administration interval can be extended compared to conventional products.

EXAMPLES Next, the present invention will be explained in detail with reference to examples, but the present invention is not limited by these in any way. Furthermore, these modification methods can be introduced into any manufacturing process of blood coagulation factors.

Example 1 Monomethyl ether polyethylene glycol
4,6-Dichloro-1,3,5-triazine: 25.0 g (0.005 mol) of polyethylene glycol monoethyl ether (average molecular weight 5000) was dissolved in 200 ml of dry benzene, and after cooling, 5.0 g of anhydrous sodium carbonate and cyanuric chloride were dissolved. 2.75g (0.015mol)
was added and stirred overnight at room temperature. After the reaction was completed, the mixture was filtered, 600 ml of petroleum ether was added to the solution, and a precipitate was obtained by suction filtration and washed with a small amount of petroleum ether. It was purified by repeating reprecipitation with dry benzene-petroleum ether three times to obtain monomethyl ether polyethylene glycol-4,6-dichloro-1,
A white powder of 3,5-triazine was quantitatively obtained.
This showed a qualitative reaction of chloride ions (AgNO ₃ ) after hydrolysis. Similarly, average molecular weight 550,
700, 2000, 10000 and 15000 polyethylene glycol monomethyl ether, polyethylene glycol monostearyl ether with an average molecular weight of 3200, polyethylene glycol stearyl ester with an average molecular weight of 2700, and cyanuric chloride,
Monomethyl ether polyethylene glycol-4,6-dichloro-1,3,5-triazine, monostearyl ether polyethylene glycol-4,6-dichloro-1,3,5-triazine, monostearyl ether polyethylene glycol-4 of each average molecular weight ,6-dichloro-1,3,5-triazine was quantitatively obtained.

Example 2 Polyethylene glycol-4-chloro-6-hydroxy-1,3,5-triazine: Polyethylene glycol (average molecular weight 6000)
Dissolve 33.6g in 150ml of dry benzene by heating, and after cooling, add 1.6g of anhydrous sodium carbonate and cyanuric chloride.
0.74 g was added and stirred at room temperature overnight. After adding 1.0 ml of water, the mixture was stirred at room temperature for 6 hours and at 40° overnight. Insoluble matter was removed by centrifugation (2000 rpm, 10 minutes), and the supernatant was distilled off under reduced pressure. The residue was dissolved in benzene while heating and evaporated. This operation was repeated two more times. The residue was dried under reduced pressure and polyethylene glycol-4-chloro-6-hydroxy-1,3,5-
A white waxy solid of triazine was obtained quantitatively. Similarly, polyethylene glycol with an average molecular weight of 1000 and 3.350 was reacted with dianuric chloride, and polyethylene glycol-4-chloro-
6-hydroxy-1,3,5-triazine was obtained quantitatively.

Example 3 Monomethyl ether polyethylene glycol
4,6-dichloro-1,3,5-triazine-modified factor factor (PEG5000--4.2mM): Factor ("Cornine", manufactured by Katsuttar) 400
Dissolve one unit of lyophilized preparation in 6 ml of distilled water to make 0.1M
Dialysis was performed in phosphate buffer (PH7.0) for 30 minutes on ice. After dialysis, the contents were diluted to 10 ml with the same buffer solution, and aprotinin ("Repulson", Mochida Pharmaceutical Co., Ltd.)
) 1.66 ml and the average molecular weight prepared in Example 1.
5000 monomethyl ether polyethylene glycol-4,6-dichloro-1,3,5-triazine
244 mg (final concentration 4.2 mM) was added, and the mixture was reacted for 3 hours under ice-cooling and stirring, and dialyzed in the same buffer at 0° for 1 hour, and further in 0.45% saline at 0° for 1 hour. After dialysis,
The contents were transferred to a vial and lyophilized to obtain a white powder of monomethyl ether polyethylene glycol-4,6-dichloro-1,3,5-triazine-modified factor (PEG5000--4.2 mM). In the same way, instead of 1.66ml of aprotinin,
Aprotinin-free PEG5000-4.2mM was prepared using 1.66ml of the above phosphate buffer.
The factor activity of PEG5000--4.2mM (without aprotinin) is 113
It was %.

Example 4 Monomethyl ether polyethylene glycol
4,6-dichloro-1,3,5-triazine modified factor (PEG5000--1.72mM and -
2.44mM): 400 units of factor as in Example 3 and Example 1
Monomethyl ether polyethylene glycol-4,6-dichloro- with an average molecular weight of 5000 prepared in
1,3,5-triazine 100 mg (final concentration 1.72 m
PEG5000--1.72mM and -2.44mM were prepared from M) and 200mg (final concentration 2.44mM), respectively.

When factor activity was measured in the same manner as in Example 3,
PEG5000 - 91.2% at -1.72mM, -2.44mM
It was 97.9%.

Example 5 Monomethyl ether polyethylene glycol
4,6-dichloro-1,3,5-triazine modified factor (PEG5000--2.1mM, 2.94mM
M and -4.2mM): Factor (“Koate”, manufactured by Katsuttar) 500
Dissolve one unit of lyophilized preparation in 10ml of distilled water to make 0.1M
Dialysis was performed in phosphate buffer (PH7.0) for 30 minutes on ice. After dialysis, divide the contents into 3 equal parts and add each part to the same buffer.
I pumped it up to 4.28ml. Monomethyl ether polyethylene glycol-4,6-dichloro-1,3,5- with an average molecular weight of 5000 prepared in Example 1
Triazine 45mg (final concentration 2.1mM), 63mg (final concentration 2.1mM)
2.94mM) and 90mg (4.2mM) were added.
React for 3 hours under ice-cooling and stirring, and add each in the same buffer.
Dialysis was performed at 0° for 1 hour and then in 0.45% saline for 1 hour at 0°. After dialysis, the contents were transferred to a vial and lyophilized to obtain a white powder of monomethyl ether polyethylene glycol-4,6-dichloro-1,
3,5-triazine modified factor (PEG5000-
-2.1mM, -2.94mM and -4.2mM).

Example 6 Monomethyl ether polyethylene glycol
4,6-dichloro-1,3,5-triazine modified thrombin (PEG5000-a-2.1mM, -
2.94mM and -4.2mM): As in Example 4, thrombin ("Thrombin-Midori", manufactured by Midori Juji Co., Ltd.) was used instead of factor factor.
Monomethyl ether polyethylene glycol-4,6-dichloro-1,3,5-triazine with an average molecular weight of 5000 prepared in Example 1 by dividing 500 units into three equal parts, 45 mg (final concentration 2.1 mM), 63 mg (final concentration 2.94 mm)
M) and 90mg (4.2mM), respectively.
PEG5000-a-2.1mM, -2.94mM and -4.2
mM was obtained.

Example 7 Polyethylene glycol-4-chloro-6-hydroxy-1,3,5-triazine-modified factor factor (PEG6000--5.0mM): Factor factor ("Cornine", manufactured by Katsuter) 400
Dissolve one unit of lyophilized preparation in 10ml of distilled water to make 0.1M
Dialysis was performed in phosphate buffer (PH7.0) for 30 minutes on ice. After dialysis, the contents were diluted to 20 ml with the same buffer and divided into 10 equal parts. To 2 ml of this, add polyethylene glycol-4-chloro-6-hydroxy-1,3,5- of average molecular weight 6000 prepared in Example 2.
60 mg of triazine (final concentration 5.0 mM) was added.
React for 6 hours while stirring on ice and add 2ml of 0.1MKH ₂ PO ₄
Stop the reaction by adding PEG6000−5.0mM
A clear aqueous solution of was obtained. Average molecular weight in a similar manner
3350 polyethylene glycol-4-chloro-6
-Hydroxy-1,3,5-triazine 33.5mg
(final concentration 50mM) to obtain PEG3350--5.0mM.

When factor activity was measured in the same manner as in Example 3,
PEG6000−-71.8% at 5.0mM, PEG3350−
-5.0mM was 77.5%.

Example 8 Monomethyl ether polyethylene glycol
4,6-dichloro-1,3,5-triazine modified factor (PEG550--10.0mM and
PEG15000--2.0mM): average molecular weight 550 and
15000 monomethyl ether polyethylene glycol-4,6-dichloro-1,3,5-triazine 11.0 mg (final concentration 10.0 mM) and 60.0 mM (same concentration)
2.0mM) to PEG500−-10.0mM and
PEG15000--2.0mM was obtained.

When factor activity was measured in the same manner as in Example 3,
PEG550−−113.8% at 10.0mM, PEG15000−
-2.0mM was 66.6%.

Example 9 Monostearyl ether polyethylene glycol-4,6-dichloro-1,3,5-triazine modified factor (PEG3200--3.0mM): Monostearyl ether polyethylene glycol with an average molecular weight of 3200 was prepared in the same manner as in Example 6. −
4,6-dichloro-1,3,5-triazine 19.2
PEG3200−−3.0m from mg (final concentration 3.0mM)
I got M. When the factor activity was measured in the same manner as in Example 3, it was 74.0%.

Example 10 Monostearyl ester polyethylene glycol-4,6-dichloro-1,3,5-triazine modified factor (PEG2700--5.0mM): Monostearyl ester polyethylene glycol with an average molecular weight of 2700 was prepared in the same manner as in Example 6. −
4,6-dichloro-1,3,5-triazine 27.0
PEG2700−−5.0m from mg (final concentration 5.0mM)
I got M.

When the factor activity was measured in the same manner as in Example 3, it was 98.1%.

Example 11 Oral administration experiment of 4.2 mM PEG5000: Approximately 4 ml of blood was collected from a healthy beagle dog (female, 10 kg). Next, PEG5000 prepared in Example 3
--400 units of the 4.2mM lyophilized product was dissolved in approximately 100ml of milk and administered orally. After administration 1, 2,
Approximately 4 ml of blood was collected after 4, 6, and 8 hours. 1/9 volume of 3.8% sodium citrate solution was added to each blood as an anticoagulant, and centrifugation was performed to fractionate plasma. Partial thromboplastin time (PTT) by adding cephaloplastin and calcium ions to plasma
Then, tissue thromboplastin and calcium ions were added to measure prothrombin time (PT).
Furthermore, the clotting time correction effect of factor-deficient plasma was measured by Quick's one-step method. The correction effect of plasma before administration is set as 100%, and the correction effect of 1/2 and 1/4 of that amount is set as 50% and 25%, respectively, and a calibration curve is created on a logarithmic graph and blood is collected at each time. The correction effect of the collected plasma was determined in relative percentage. In addition, prepared in Example 4
Similar experiments were conducted with PEG5000--1.72mM and -2.44mM. Furthermore, as a control experiment,
400 units of unmodified factor was orally administered to dogs in a similar manner and the plasma was analyzed. The results are shown in FIGS. 1 to 5. No particular side effects were observed in the experiments with the dogs used.

Example 12 Oral administration experiment of PEG5000: PEG5000 prepared in Example 5--2.1mM
177 units of PEG5000--2.94mM and 177 units of PEG5000-4.2mM were mixed;
Dogs were orally administered in the same manner as in Example 10, blood samples were collected over time, and plasma was analyzed. Furthermore, as a control experiment, an experiment was conducted in which 500 units of unmodified factor was orally administered in the same manner. The results are shown in Figure 6~
It is shown in FIG. No particular side effects were observed in the experiments with the dogs used.

[Brief explanation of drawings]

Figure 1 shows the animal experiment conducted in Example 11.
Indicates PTT. Γ-Γ: unmodified factor (no aprotinin)
Treated dog, ●...●: PEG5000--4.2mM (no aprotinin) dosed dog, ●-●: PEG5000--
Dog administered with 4.2mM (containing aprotinin) Figure 2 shows the PT of the animal experiment conducted in Example 11.
shows. Γ-Γ: unmodified factor (no aprotinin)
Treated dog, ●...●: PEG5000--4.2mM (no aprotinin) dosed dog, ●-●: PEG5000--
Dog administered with 4.2mM (containing aprotinin) Figure 3 shows the factor activity in the animal experiment conducted in Example 11. Γ-Γ: unmodified factor (no aprotinin)
Treated dog, ●...●: PEG5000--4.2mM (no aprotinin) dosed dog, ●-●: PEG5000--
Dog administered with 4.2mM (containing aprotinin) FIG. 4 shows the factor activity in the animal experiment conducted in Example 11. Γ-Γ: unmodified factor (no aprotinin)
Treated dog, ●...●: PEG5000--4.2mM (no aprotinin) dosed dog, ●-●: PEG5000--
Dog administered with 4.2mM (containing aprotinin) FIG. 5 shows the factor activity in the animal experiment conducted in Example 11. Γ-Γ: unmodified factor (no aprotinin)
Treated dog, ●...●: PEG5000--4.2mM (no aprotinin) dosed dog, ●-●: PEG5000--
Dogs administered with 4.2mM (containing aprotinin) Figure 6 shows the animal experiment conducted in Example 12.
Indicates PTT. Γ-Γ: unmodified factor (no aprotinin)
Administration dog, ●-●: PEG5000--2.1mM, -2.94
Figure 7 shows the PT of the animal experiment conducted in Example 12.
shows. Γ-Γ: unmodified factor (no aprotinin)
Administration dog, ●-●: PEG5000--2.1mM, -2.94
Dogs dosed with mM and -4.2 mM (no aprotinin) FIG. 8 shows the factor activity of the animal experiment conducted in Example 12. Γ-Γ: unmodified factor (no aprotinin)
Administration dog, ●-●: PEG5000--2.1mM, -2.94
Dogs treated with mM and -4.2mM (no aprotinin)

Claims (1)

[Claims] 1. The amino acid side chain of a blood coagulation factor has the general formula RO(CH ₂ CH ₂ O) _o -A- (wherein R is a hydrogen atom, an alkyl group or an alkanoyl group, and n is an integer. , A represents an active residue capable of binding to an amino group, and the molecular weight of RO (CH ₂ CH ₂ O) _o is 200 to 20,000) is bound to a new blood coagulation factor. derivative. 2. The novel blood coagulation factor derivative according to claim 1, wherein A is a triazine ring having a halogen atom or a hydroxyl group on the ring. 3. The novel blood coagulation factor derivative according to claim 1 or 2, wherein R is a methyl group. 4. The novel blood coagulation factor derivative according to claims 1 to 3, wherein the molecular weight _of RO( _CH2CH2O ) _o is 1000 to 10000. 5. The novel blood coagulation factor derivative according to claims 1 to 4, wherein the blood coagulation factor is any one of factor No. 6 Blood coagulation factors and the general formula RO (CH ₂ CH ₂ O) _o -A-X (wherein, R is a hydrogen atom, an alkyl group or an alkanoyl group, n is an integer, and A is a bond to an amino group) blood coagulation, characterized by reacting a conjugate of polyethylene glycol and a coupling agent, represented by RO (CH ₂ CH ₂ O ₎ , whose molecular weight is between 200 and 20000). The amino acid side chain of the factor is represented by the general formula RO(CH ₂ CH ₂ O) _o -A- (wherein, the molecular weights of R, n, A and RO(CH ₂ CH ₂ O) _o are the same as above). A method for producing a novel blood coagulation factor derivative having a group bonded thereto. 7. The amino acid side chain of the blood coagulation factor has the general formula RO(CH ₂ CH ₂ O) _o -A- (wherein R is a hydrogen atom, an alkyl group or an alkanoyl group, n is an integer, and A is an amino group). RO (CH ₂ CH ₂ O) _o has a molecular weight of 200 to 20,000) and contains a novel blood coagulation factor derivative containing a group that can be bonded. A characteristic blood coagulation promoter. 8. The blood coagulation promoter according to claim 7, wherein the molecular weight _of RO( _CH2CH2O ) _o is 1000 to 10000. 9. The blood coagulation promoter according to claim 7 or 8, wherein the administration route is oral.