Method of producing hydrazone conjugate

专利摘要:
Immunoglobulin conjugates formed by reaction of an antineoplastic indole-dihydroindole vinca alkaloid containing a hydrazide group attached at C-3 or C-4 with an oxidized glycoprotein containing aldehyde groups, are provided.
公开号:SU1561825A3
申请号:SU874202656
申请日:1987-05-22
公开日:1990-04-30
发明作者:Коулмен Лагузза Беннет；Линн Николс Синтиа
申请人:Эли Лилли Энд Компани (Фирма)；
IPC主号:

专利说明:

This invention relates to a process for the preparation of new conjugated derivatives of vinblastine alkaloids,
 r.f., cii2-CH3
and the formula
H
he
namely, the hydracon conjugate of the general formula
., cii
H
| pC-OSSN H I 6
.нCHjO
-CHj-CH, j CP ,, ORi he
CHjr COR
where r
ъ
m
OSI, or - NHN CH-rpynna; Hi - COClI9.CHe-CONHN CH-rpynna 1-10, and R cannot be - NHN CH-, when 11 / - (CO (CH2) ZCONHN-CH-, and R cannot be CO (CK.t) 2CONHN -CH- when R is NHN-CH-, GP is the residue of an immunoglobulin or its fragment,
with antitumor activity.
The purpose of the invention is to obtain new conjugates of hydralone, which have pharmacological advantages over conjugated analogues.
PRI me R 1. Preparation of 4-deoxy-4-deacetyllerosidin-3-carboxyhydrazide-N2-succinimide.
The solution is prepared from 1320 microns of 4-deoxy-4-deacetyllerosidin-3-carboxigidrazide in 25 ml of pyridine. 175 mg of succinic anhydride is added to the solution and the reaction mixture is stirred at room temperature under nitrogen atmosphere for about 24 hours. Then the petting components are removed from the reaction mixture in vacuum, and the resulting precipitate absorbs methylenedichloride in combination with a sufficiently n-glutaroyl 4-deacetyl of methanol to completely dissolve the resulting precipitate. The organic layer is washed twice with an equal volume of water and then dried. When the volatile components are evaporated in vacuum, a precipitate is obtained containing M2-succinoyl 4 - deoxy-4-deacetyllerosidine-3-carboxyhydrazine, which is dissolved in
25 ml of pyridine, to which is added to acetyl-UHB-3-carboxyhydrazide-K 2350 mg of acetic anhydride. As a result, a mixed anhydride of succinic and acetic acids is formed, which simultaneously cyclizes, yielding 4-deoxy-4-deacetyllerosidine-45H-carboxyhydrazide-N2-succinimide. The reaction mixture is evaporated to dryness in vacuo, and the resulting precipitate is dissolved in methylene dichloride. The methylene dichloride extract is washed twice with 50 equal volumes of water and then dried. By evaporation of methylene dichloride, 4-deox C-4 deacetyl rosidin-3-carboxy is obtained,
glutarimide. The compound indicated is purified by chromatography on silica gel using the indicated solvent system. 460 mg of N-g tarimide derivative is obtained, which has the following physical characteristics:
Mass spectrum, п / е 864 (C48H60N
IR (CHS19): 3475; 1714 and 1616 cm-1.
pKa (66% aqueous solution of DMF DMF): 5; 1.7; 12.9.
NMR (CDC13): 9.9b; 8.91; 8.04;
hydrazide-M2-succinimide, which is 55,52,52; 7.14; b, 58; b, 08; 5.83; 4.15;
Computed on a silica gel column with ethyl acetate, containing increasing amounts of methanol (0-50%) as eluent. Fractions that
3.78; 3.69; 3, S; 2.8b; 0.93; 0.89. Sulfate salt is prepared according to the indicated method. From 290 mg of the starting material, 210 mg of salt is obtained.
na; 5 - ay
15618254
(as established by thin layer chromatography) contain the desired succinimide derivative, combined and evaporate in vacuo a solid. 190 mg of 4-deoxy-4-deacetyl-leurosidine-N-3-carboxyhydrazide-H-succinimide were obtained with the following physical characteristics:
IR spectrum (in chloroform): 1736; 1615 and 1572.
NMR (CDC13): 10.90; 7.88; 7.49;
7.12; 6.48; 6, Ob; 5.80; 4,10; 3.76; 3, 6; 2.82; 0.93: 0.85.
The sulfate salt is obtained by dissolving the free base in anhydrous ethanol at 0 °. Then the pH is adjusted to 3.9 using a freshly prepared solution of 2% sulfuric acid in ethanol. By evaporation of the reaction mixture to dryness, 4-deoxy-4-deacetylleurozidine-3-carboxyhydrazide-N2-succinimide sulfate is obtained.
Following this procedure, acetylvinblastine hydrazide (2.4 g) is dissolved in 125 ml of pyridine, to which 385 mg of glutaric anhydride is added. Received so
five
VLB-3-carboxyhydrazide is isolated according to this procedure and purified by silica gel chromatography using the same solvent system as described above. The resulting glutamine compound interacts with acetic anhydride to form a mixed anhydride, which simultaneously cyclizes to yield 4-desglutarimide. The compound indicated is purified by chromatography on silica gel using the indicated solvent system. 460 mg of the n-gluari-imide derivative is obtained, which has the following physical characteristics:
Mass spectrum, п / е 864 (C48H60N60,)
IR (CHS19): 3475; 1714 and 1616 cm-1.
pKa (66% aqueous solution of DMF DMF): 5; 1.7; 12.9.
NMR (CDC13): 9.9b; 8.91; 8.04;
7.52; 7.14; b, 58; b, 08; 5.83; 4.15;
3.78; 3.69; 3, S; 2.8b; 0.93; 0.89. Sulfate salt is prepared according to the indicated method. From 290 mg of the starting material, 210 mg of salt is obtained.
Following this method, but replacing 4-deacetyl U1, B-3-car boxyhydrazide on 4-deoxy-4-deacetylleurozidine-J-carboxyhydrazide, using acetyl chloride instead of acetic anhydride, you get M2-succinoyl 4-deacetyl- VLB-3-carboxyhydrate, When chromatographing a succinimide product, two fractions are released, one of which is the expected 4-des-acetyl, UHB-3-carboxy-hydrazide-n-succinimide, and the other is its 4-acetyl-derivative, which forms as a by-product product during the cyclization process. 4 dezacetyl HBB-3 carboxyhydrazide-L2-succinimide has the following physical characteristics:
Mass spectrum, (,).
IR spectrum (KVg): 3480; 1734 and 1616.
NMR (C1) C13): 9.69; 9.47: 8.22; 7.49; 7.12; 6.53; 6.07; 5.82; 5.71; 4.01; 3.77; 3.63; 3.59; 2.84; 2} 82 and 0.89.
Sulfate salt get in the specified method. Yield 250 mg from 350 mg of starting material.
II p and m e r 2. Prepare a solution by dissolving 2 $ 5 g of 4-deacetyl-4-hemisuccinate (4-succinoyl-UHB) in 50 ml of chloroform. 870 mg of N-methylmorpholine is added. When the dissolution is complete, the reaction mixture is cooled in an ice bath and a stream of nitrogen is supplied. 985 mg of isobutyl chloroformate was added and the reaction mixture was stirred at 0 ° C for 45 minutes. 1 ml of anhydrous hydrazine is added and the reaction mixture is stirred for about 10 minutes. Then, the reaction mixture was extracted with water, the aqueous layer was separated and discarded, and the organic layer was dried. Evaporation of the solvent gives a residue containing 4-deacetyl L B 4-hemisuccinate hydrazide. A solution of the residue in methylene chloride is chromatographed on silica gel using a mixture of ethyl acetate and methanol (100: 0 - 50:50) as eluent. Corresponding fractions give 950 mg of purified hydrazide 4-deacetyl U1, B-4-hemisuccinate; .
The sulfate salt is obtained by dissolving the base in anhydrous ethanol (25 ml), adjusting the pH to about 4 with a 2% solution of sulfuric acid in ethanol (24.5 g of anhydrous ethanol, 0.5 g of 18 M) and removing solvents in vacuo .
IR spectrum: 3400; 3009; 1738; 1680 and 1616 cm (.
NMR (CDC1,): 9.9; 8.35; 8.05; 7.55; 7.10; 6.85; 6.10; 5.85; 5.45; 5.30; 3.80; 3.70; 3.60; 2.70; 0.90; 0.85. Offered conjugates include
oxidized glycoproteins, preferably immunoglobulins, and of this class, preferably monoclonal antibody (MoAB), which is a gamma globulin, such as IgC
5 or IgM. Immunoglobulin fragments (ig) containing carbohydrate, as well as the original immunoglobulin from which these fragments are obtained, can be used to form new
Q conjugates of the same type.
A preferred glycoprotein class (immunoglobulins) is compounds that react or at least recognize anti-5 genes on the target cell, i.e. they have antigen recognition properties. Particularly preferred are those glycoproteins that recognize antigens on the surface
0 target cells.
The preferred form of antibody for use in the invention is an immunoglobulin, which is gamma globulin, in particular
5 preferred are IgC, IgA, IgE and IgM species.
Preferred conjugates are those prepared from monoclonal antibodies, especially those recognized by human cancer cells, such as adenocarcinoma. scaly cell carcinoma, transitional cell carcinoma, melanoma, neuroblastoma, small cell carcinoma
5 noma, leukemia, lkmphoma and sarcoma.
The glycoprotein (HP) of this type can be oxidized with periodate or another suitable oxidizing agent so that the bond between adjacent diols
m in the surface carbohydrate is broken and aldehyde groups are formed on both sides of the original bond
I,
ns it
ns - he
sno
tp
Part of the carbohydrate structure is a dialdehyde product.
The number of dialdehyde units produced is the result of the following factors; the amount of periodate used, the general conditions of oxidation reactions (for example, time, temperature, solvent, concentration, etc.) | the number of neighboring diodes of carbon units present in the protein and their availability for periodate reagent.
Alternatively or in combination with periodate, enzymatic oxidation can be used, for example, galactose oxidase. It is a more selective and limiting reagent that catalyzes the conversion of the 6-OH group of galactose residues in the carbohydrate chains of glycoprotein to aldehyde groups. This 6-CHj OH-rpynna must be unsubstituted in order to ensure successful oxidation. Unmasking a blocking group, such as a group in which there is a sialic acid residue, can be provided by the Neurominidase enzyme. The number of aldehyde residues obtained is a function of many parameters.
Aldehyde-containing glycoproteins (OCH) m-GP are then conjugated to vinca-hydrazide.
The number m is a function of the number of aldehyde groups arising from the described oxidation. Conjugates containing 25 vincaostats per antibody were detected. The mean value (of vino-residues per antibody) is about 1 - 10 (i.e. - 10), the preferred value is about 4 - 10.
Conjugation of vincahydrazides with oxidized glycoproteins is carried out according to standard procedures. A solution of vinca compound in a solvent that is miscible with water, such as dimethylformamide, is added to a cooled, buffered aqueous solution of an oxidized glycoprotein. Preferred temperatures - 8 ° C, usually used buffer 0.1 n. sodium acetate. The reaction is best carried out in the dark and in an inert atmosphere. The reaction usually ends in 10 to 24 hours, and the resulting conjugate can be purified by standard methods, such as siadex chromatography.
The following is a description of the oxidation and conjugation of specific MoAb.
PRI me R 3. Conjugation with X-63AG8-S1 MOAB.
Prepare a solution by dissolving 200 mg of X-63AG8-S1 (1 ml of approximately 150000, 1, 34xHg / mol; cell line deposited as ATCC 11B9; collection of American type cultures, 12301 Parklawn Drive, KockvilJe MD 20852) in 20 ml of buffer 0, 1 M sodium acetate, pH 5.6 (29.3 g sodium acetate, 2.44 ml of acetic acid plus enough sterilized water to make 4 liters of buffer). The solution is stored at 0 ° C overnight (about 10% of protein not dissolves). 685 mg of sodium metaperiodate is added in one portion with vigorous stirring.
The mixture is stirred for 21 minutes at 0 ° C in the dark and then quenched by adding a 5-fold excess (for the full amount of periodate) ethylene glycol (1.28 ml of a 12.5 M solution of ethylene glycol in sterile water). The new mixture is stirred at 0 ° C. for 5 minutes in the dark and then centrifuged to obtain a clear supernatant and a white precipitate. The supernatant was loaded onto a Sephadex G25 gel (medium grain) and the product was eluted with the same sodium acetate buffer. The eluate is checked by UV irradiation at 280 mm. The period is washed away from the column and discarded. The concentration of the oxidized product is evaluated in each fraction of the eluate at 279 nm. Yield 96%. The second experiment carried out also gives a 96.5% yield.
Solutions of the eluate, containing 4.72 and 4.02 mg / ml of the oxidized product, are mixed and 0.1N buffer is added. sodium acetate to obtain a final protein concentration of 2.77 mg / ml (total volume 69 ml). The solution is cooled to about 0 ° C.
A solution of 4-deacetyl-U1., B-3-carboxy-sigidrazide in DMF (5.6 ml and 53.7 mg / ml solution) is added dropwise to the cooled MoAb buffer solution. The reaction vessel is washed with nitrogen and closed. The reaction mixture is stirred under cooling and in the dark with a magnetic stirrer for 24 hours. Then the reaction vessel. open and centrifuge the clear pale yellow reaction mixture. The supernatant is chromatographed on Sephadex G25 gel;
Antibody
The average number of moles of vinka per mole of antibody in the final conjugated product
ten
15
balanced to pH 7.4, Table1
phosphate-saline buffer (0.01 M, 0.15 M NaCl). which is also used as eluent.
The conjugate (formed by a hydrazone bond majeu by a 3-carboxyhydrazide group and an aldehyde group in MoAb) is eluted first, followed by elution of unreacted 4-deacetyl-VLB-3-carboxyhydrase. The yield of the resulting conjugate (from 4 columns) is .73 mg in 224 ml (90% yield). The conjugate contains about 6 mol of 4 deacetyl-ghB 3. -carboxyhydrazone per mole of X-63AG8 - S1 MoAb.
The second experiment was carried out using 200 mg of KS 1/4 MoAb capable of recognizing cell surface antigens (human adenocarcinoma) in 20.0 ml of acetate buffer and the same amount of periodate and ethylene glycol as in Example 3. Experience gives 176 mg of oxidized MoAb in 39.9 ml buffer after chromatography (88% yield). Conjugation with 274 mg of 4-deacetyl-VLB-3-carboxyhydrazide in acetate buffer, pH 5.6, gives 146 mg (83% yield) of the conjugate containing 30 used in the calculation of the average.
20
25
11,285.14
11А8
11B2
13H1
14.95.55
17J12
2A11
324.9.4
4D12
4E8.11
6D2
G9
L1ks
L2ks
L4ks
L6ks
ZCE-025
ten
4.2 (3)
5.7 (4)
5.5 (6)
4.1 (1)
6.2 (4)
4.2 (1)
6.0 (1) 3.9 (2)
4.3 (6)
4.4 (3)
5.1 (4)
6.6 (2)
5.8 (7)
6.4 (2)
4.5 (4) 4.25 (2)
11.9 (11)
Note. The number in parentheses indicates the number of experiments; a non-group of gats containing about 27 vinka-groups per antibody was detected in this group.
about 7.5 moles of 4-cesacetyl B-3-carboxyhydrase per mole of KS 1/4. According to this method, the conjugate is obtained by reacting 4-desacetyl-VLB-3-carboxyhydrazide with 35 aldehyde groups formed by oxidation of surface carbohydrates 9.2.27 of the glycoprotein MoAb capable of recognizing antigenic properties on the surface of human melanoma cells of the 40th century. The experiment with the use of 200 mg of MoAb, oxidized 685 mg of sodium metaperiodate in acetate buffer at pH 5.6 (0.1 M), yields an aldehyde-containing oxidized 9.2.27 MoAb with a 92% yield (184 mg). This product is conjugated to 4 decachetyl U1., B-3-carboxyhydrazide (279 mg). The final yield of poly-4-deacetyl U1, B-hydrazone is oxidized. In this group, one conjugate was found containing about 27 vink-groups per antibody.
Conjugates can also be prepared with immunoglobulin Ig fragments, IgM monomer or other Ig monomers containing carbohydrate obtained from the original antibody by, for example, cleavage with a proteolytic enzyme or reductive digestion. Preferred proteolytic Enzymes for producing these fragments are pepsin and papain.
A study of the conjugates of the invention can be carried out using a known technique such as Athens on chromatographs. The effect of the conjugate can be evaluated by counting i
aldehyde containing 9.2.27-conjugate number of viable cells after treatment with a conjugate of a suspension of tumor cells or by measuring the absorption of tritiated uridine. The concentration of protein and drug preyugate as a solution in phosphate-saline buffer is 91%.
The product contains about 4.9 mol of the initial wine per mole 9.2.27.
According to the procedure described in example 3.55 paRat is determined by measuring
as presented in table. 1, several other antibody conjugates were prepared using 4-deacetyl-VLB-3-carboxigidrazide as the remnant.
optical density of konghat solutions at two wavelengths, for example, 270 and 279 nm, and comparing the values obtained with
The average number of moles of vinka per mole of antibody in the final conjugated product
five
0 used in calculations
0
five
11,285.14
11А8
11B2
13H1
14.95.55
17J12
2A11
324.9.4
4D12
4E8.11
6D2
G9
L1ks
L2ks
L4ks
L6ks
ZCE-025
ten
4.2 (3)
5.7 (4)
5.5 (6)
4.1 (1)
6.2 (4)
4.2 (1)
6.0 (1) 3.9 (2)
4.3 (6)
4.4 (3)
5.1 (4)
6.6 (2)
5.8 (7)
6.4 (2)
4.5 (4) 4.25 (2)
11.9 (11)
used when calculating the average.
Note. The number in parentheses indicates the number of experiments that were not used to calculate the average.
In this group, one conjugate was found containing about 27 vinka groups per antibody.
Conjugates can also be prepared with immunoglobulin Ig fragments, IgM monomer or other Ig monomers containing carbohydrate obtained from the original antibody by, for example, cleavage with a proteolytic enzyme or reductive digestion. Preferred proteolytic Enzymes for producing these fragments are pepsin and papain.
A study of the conjugates of the invention can be carried out using a known technique such as Athens on chromatographs. The effect of the conjugate can be evaluated by counting i
the numbers of viable cells after treatment with a suspension of a tumor cell suspension or by measuring the uptake of tritiated uridine. The concentrations of protein and drug preparation are determined by measuring
the optical density of the conjugate solutions at two wavelengths, for example, 270 and 279 nm, and comparing the obtained values with the values for
free drug and unconjugated immunoglobulin at the same wavelengths. The conjugate can also be evaluated in vivo against human tumor xenografts in atypical mice.
New conjugates are effective over a wide dosage range. For example, for treating adults with cancer, the doses are usually in the range of 0.01–10 mg / kg (the remainder of the distillery), most often in the range of 0.03–9 mg / kg. However, it should be borne in mind that The amount of co-conjugate taken is determined by the doctor depending on the condition and course of treatment.
The following tables indicate the effect of the proposed conjugates in the treatment of tumors.
In tab. Figure 2 shows the minimum effective dose (Winka equivalent) in melanoma models of the conjugate of 9.2.27-deacetyl-VLB-hydrazide.
Table 2

and
ohm
ME, mg / kg content of wine
SO, 0625 0.375 0.375 0.375 0.185
Note. N.E.D. - the minimum effective dose that demonstrates 50% suppression of tumor growth for 28 days, conjugate 9.2.27-deacetyl L, B-hydrazide in ne

ABOUT
РЗИ CLA НТ-29 FA1M LS174 РЗИ CLA РЗИ СТА РЗИ CLA РЗИ CLA. LS174 LSI 74
The calculation of the content of the Vinca for three times intravenous administration (2, 5 and 8 days after the implantation of the tumor).
In tab. Figure 3 shows the minimum effective dose (Winka equivalent) in xenograft models of adenothoid carcinoma of the KS1 / 4-deacetyl-VLB-hydrazide conjugate.
T a b l and c a 3

VL M.E.D. mg / kg content of wine
RZ-ISED - carcinoma of the lung
human: 0.0625 NT-29 - colorectal carcinoma 0.25 NT-29 VTE - colorectal carcinoma (), 25
 M.E.D. -minimal effective dose, which demonstrates 50% suppression of tumor growth, conjugate KS1 / 4-deacetyl-UHB-hydrazide in terms of the content of vinka for three intravenous administration (2, 5 and 8 days after tumor implantation).
 The HT-29VTE is an in vivo selected cell line variant resistant to vinka alkaloids.
In tab. Figure 4 shows the minimum effective dose of monoclonal antibody 4 conjugates of deacetyl-U1, B-hydrazide in models of human tumors in mice. i
Table4
Oj05 0.0625 0.5 0.3 0.125 0.125 1.2 0.6, 5 0.5
 Same as in table. 2 and 3. Winky-content of the conjugate - 4-deacetanl VLB-hydrazide.
 REI CLA - lung adenocarcinoma; HT-29 - colon adenocarcinoma; FADIi - pharyngeal scaly cellular carcinoma; LSI 74 - colon adenocarcinoma; T222 - cellular carcinoma. The animals were injected conjugate three times in the period
2 8 days after implantation, tumor. Tumors were measured 28 days after implantation, Inhibition was determined in relation to animals treated with PbS as controls.
 The same protocol was used as in 1), but the dose was administered on days 3, 6 and 9 and tumors were measured after 2,
3 or 4 weeks.
In the experiment, the nude mice were infected with the CLA cells of the human lung adenocarcinoma by subcutaneous administration on day 0. On the 2nd, 5th and 8th day after this, groups of four or five animals were treated by either intravenous injection or conjugate of the described vinka derivative . On the 28th day, a tumor was measured.
Medicine
T
Dose, mg / kg I Percent inhibition
mg / kg of wine or wine content.
Continuation of table 4
and the percent inhibition was determined by comparing the tumor mass of the treated groups of animals with untreated ones. control animals. The dose is expressed in mg / kg of the content of vinka in the conjugate.
The results of the experiment are given in table. five.
In tab. 5 shows experience with adenocar -. RZI CLA.
T
Table5
I Percent Inhibition
In tab. 5 DAVLBtidznd is a 4-deacetylvinblastin-3-carboxyhydrazide, as described in British patent application f OB 2 090 83 / A, while the conjugate DAVLB-hydrazide - KS1 / 4 is a conjugate prepared according to the procedure described in example 3,
As can be seen from the above data, basically complete suppression of tumor growth was observed at dosage levels of the conjugate of 1-0 mg / kg, whereas at the same level, the free DAVLB-hydrazide exhibited only minimal effect. Thus, the experiment shows that the drug - an immunoglobulin conjugate according to the invention has obvious advantages compared to the free drug in the co-doped form.
Similarly, a comparison of the same conjugate with a conjugate with the same antibody, but prepared as described in the USSR patent If 1362404, i.e. the succinate conjugate, known as conjugate-4-deacetyl winbetin-4-hemisuccinate with antibody KS1 / 4, was carried out according to the same scheme as described in Table 5, but in human colorectal adenocarcinoma. Measurement of the tumor produced on the 35th day. The results are shown in Table. B.
In Table 6, human colorectal adenocarcinoma HT-29 is given.
T a b l and a ab
Medicine
Dose, mg / kg
Percent Inhibition


10.0
5.0 2.5, 1.25
3.0
1.5
0.73 0.375
25
18
ABOUT
100 97
100 100
mg / kg of wine or wine content. As can be seen from the obtained data, the hemisuccinate conjugate at doses up to 10 mg / kg of distillery equivalent showed, at best, only moderate activity, which did not come close to
complete suppression of tumor growth, and the proposed hydrazide conjugate showed mainly the suppression of tumor growth at doses of 3 - 0.375 mg / kg.
Thus, the experiment shows a greater biological activity of the proposed conjugates compared with the conjugates of the compared vink and antibodies conjugated in a different way. Conjugates exhibit moderate toxicity, but have a greater therapeutic index than known drugs and conjugates used to treat cancer, and can be effectively used to treat cancer patients.

权利要求:
Claims (1)
[1]
Invention Formula
The method of obtaining the hydrazone conjugate of the general formula
HE
-ify-CHrCHj
and
GP
SNg-CH3 ORi
i Gon
ch3 | , t
COR
where R is OCH3 or -CH CH-group;
R, - N. COCHiCH2 -CONHN CK-rpynna;
m 1-10, and R cannot be when R, is CO (CH2) 2 CONHN is CH-, and K cannot be CO (CHZ) 4CONHN-CH-, when R is NHN-CH-, GP is immunoglobulin residue or fragment thereof,.
characterized in that the hydrazine vinca of common Aormula II
50
SNg-CH3
CIS-CH3
CH30
17 156182518
where R is the NHNHj group or OCH3, H, when R is
or CO (CHZ) 2C () NHNH2, with NHNH2
Rjrfe is NHNH2, when exposed to aldeR, - CO (CH2) 2 CONHH2 and R, a non-hydroxyl group of oxidized immuno is CO (CH2) 4 CONHH2, globulin or its fragment.

类似技术:

---

公开号 | 公开日 | 专利标题

---

SU1561825A3|1990-04-30|Method of producing hydrazone conjugate

---

US5585499A|1996-12-17|Cyclopropylbenzindole-containing cytotoxic drugs

---

US7528130B2|2009-05-05|Cytotoxic agents comprising taxanes and their therapeutic use

---

US6630579B2|2003-10-07|Cytotoxic agents comprising modified doxorubicins and daunorubicins and their therapeutic use

---

EP0328446B1|1994-07-20|Conjugates of vinca derivatives containing a detergent chain in position C-3

---

RU2116087C1|1998-07-27|Binding agent for bioactive preparations, method of its synthesis and pharmaceutical compositions

---

EA010508B1|2008-10-30|Method of targeting specific cells population using cell-binding agent maytansinoid conjugates linked via a non-cleavable linker, said conjugates, and methods of making said conjugates

---

HU191647B|1987-03-30|Process for producing conjugates of vinca alkaloides and immunoglobulines

---

KR20190004662A|2019-01-14|Compounds comprising cleavable linker and use thereof

---

HU206377B|1992-10-28|Process for producing cytotoxic vinc-dimer conjugates and pharmaceutical compositions comprising same

---

DK174739B1|2003-10-13|Carrier-drug conjugates and their preparation

---

EP0206666A2|1986-12-30|Hydrazide imide derivative of indole-dihydroindole alkaloids

同族专利:

---

公开号 | 公开日

---

JPS62287000A|1987-12-12|

---

NZ220441A|1989-09-27|

---

EP0247792A3|1989-07-12|

---

CN87104430A|1988-05-11|

---

PT84932A|1987-06-01|

---

AU7334987A|1987-12-03|

---

KR870011132A|1987-12-21|

---

CA1288896C|1991-09-10|

---

EP0247792A2|1987-12-02|

---

IL82579D0|1987-11-30|

---

HUT44033A|1988-01-28|

---

PT84932B|1990-02-08|

---

DK260687D0|1987-05-22|

---

AU600727B2|1990-08-23|

---

DK260687A|1987-11-28|

---

ZA873600B|1988-12-28|

---

KR900006908B1|1990-09-24|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

FI820020L|1981-01-12|1982-07-13|Lilly Industries Ltd|IMMUNOGLOBULINKONJUGATER|

---

GB2137210B|1983-03-30|1986-11-19|Lilly Industries Ltd|Immunoglobulin conjugates|

---

US4675400A|1985-06-17|1987-06-23|Eli Lilly And Company|Bifunctional derivatives of 4-desacetyl indole-dihydroindole alkaloids|

---

AT89567T|1985-11-12|1993-06-15|Wallone Region|VINBLASTIN CONJUGATES AND ITS DERIVATIVES, METHOD FOR THE PRODUCTION THEREOF AND PHARMACEUTICAL COMPOSITIONS CONTAINING THESE CONJUGATES.|GB8728856D0|1987-12-10|1988-01-27|Bradwell A R|Tumour-locating agents & method of locating tumours|

---

IL89043D0|1988-01-27|1989-08-15|Lilly Co Eli|Antibody conjugates|

---

US5006652A|1988-08-08|1991-04-09|Eli Lilly And Company|Intermediates for antibody-vinca drug conjugates|

---

US5010176A|1988-11-10|1991-04-23|Eli Lilly And Company|Antibody-drug conjugates|

---

ZA902519B|1989-04-04|1991-12-24|Lilly Co Eli|Pharmaceutical formulations|

---

WO1991016324A1|1990-04-25|1991-10-31|The Upjohn Company|Novel cc-1065 analogs|

---

JP2000506494A|1995-10-18|2000-05-30|メルクエンドカンパニーインコーポレーテッド|Complexes useful for treating benign prostatic hyperplasia|

---

DE60231868D1|2001-04-24|2009-05-20|Purdue Research Foundation|FOLAT MIMETICS AND THEIR FOLAT RECEPTOR BINDING CONJUGATES|

---

KR20040106547A|2002-05-15|2004-12-17|엔도사이트, 인코포레이티드|Vitamin-mitomycin conjugates|

---

AR042942A1|2003-01-27|2005-07-06|Endocyte Inc|CONJUGATES OF DRUG ADMINISTRATION OF VITAMIN RECEPTORS UNION|

---

US8288557B2|2004-07-23|2012-10-16|Endocyte, Inc.|Bivalent linkers and conjugates thereof|

---

WO2006101845A2|2005-03-16|2006-09-28|Endocyte, Inc.|Synthesis and purification of pteroic acid and conjugates thereof|

---

JP5475992B2|2005-08-19|2014-04-16|エンドサイト，インコーポレイテッド|Multidrug ligand conjugate|

---

EP1948240A2|2005-08-19|2008-07-30|Endocyte, Inc.|Ligand conjugates of vinca alkaloids, analogs and derivatives|

---

WO2008101231A2|2007-02-16|2008-08-21|Endocyte, Inc.|Methods and compositions for treating and diagnosing kidney disease|

---

CN101678124A|2007-03-14|2010-03-24|恩多塞特公司|Binding ligand linked drug delivery conjugates of tubulysins|

---

US9877965B2|2007-06-25|2018-01-30|Endocyte, Inc.|Vitamin receptor drug delivery conjugates for treating inflammation|

---

CN101784565B|2007-06-25|2014-12-10|恩多塞特公司|Conjugates containing hydrophilic spacer linkers|

---

WO2013126797A1|2012-02-24|2013-08-29|Purdue Research Foundation|Cholecystokinin b receptor targeting for imaging and therapy|

---

US20140080175A1|2012-03-29|2014-03-20|Endocyte, Inc.|Processes for preparing tubulysin derivatives and conjugates thereof|

---

MX2015004757A|2012-10-16|2015-07-17|Endocyte Inc|Drug delivery conjugates containing unnatural amino acids and methods for using.|

---

CN108948128B|2018-07-02|2021-10-01|中国科学技术大学|Catalyst-free hydrazone-linked polypeptide or protein chemical modification method based on electron-deficient benzaldehyde|

法律状态:

优先权:

---

申请号 | 申请日 | 专利标题

---

US86718386A| true| 1986-05-27|1986-05-27|

---