WO2005049635A2 - Humaner monoklonaler antikörper mit fettsenkender wirkung - Google Patents
Humaner monoklonaler antikörper mit fettsenkender wirkung Download PDFInfo
- Publication number
- WO2005049635A2 WO2005049635A2 PCT/DE2004/002503 DE2004002503W WO2005049635A2 WO 2005049635 A2 WO2005049635 A2 WO 2005049635A2 DE 2004002503 W DE2004002503 W DE 2004002503W WO 2005049635 A2 WO2005049635 A2 WO 2005049635A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- seq
- polypeptide
- ldl
- polypeptide according
- acid sequence
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
Definitions
- the invention relates to a purified polypeptide (SAM-6.10) and its use in combination with conventional auxiliaries and / or
- Thrombus formation Atherosclerosis with its complications (coronary heart disease, heart attack, peripheral arterial disease, stroke) is still the most common cause of death in the western world. It is estimated that well over half of all financial resources available for medical care are spent on the consequences of arteriosclerosis. In order to clarify the causes of arteriosclerosis, various theories have been developed, the lipid theory being the most respected.
- Fats such as B. cholesterol
- Fats are neither soluble in water nor in blood fluid.
- the fats are bound to certain protein bodies (proteins) as soon as they are in the blood.
- proteins proteins
- lipoproteins of the plasma are high-molecular water-soluble complexes, which consist of lipids (cholesterol, triglycerides, phospholipids) and apolipoproteins.
- lipids cholesterol, triglycerides, phospholipids
- apolipoproteins The lipoprotein LDL-cholesterol containing cholesterol causes arteriosclerosis and also becomes "bad".
- Cholesterol called, the oxidized form of LDL cholesterol is even more dangerous for the body.
- Crystalesterol is ubiquitously synthesized in the body and is an essential component of cell membranes and lipoproteins
- the sterol ring of the cholesterol molecule can no longer be broken down; Cholesterol is converted to bile acid in the liver or excreted unchanged in the intestine via the bile.
- LDL low-density lipoproteins
- Triglycerides are esters of glycerol with three fatty acid residues. Analogous to cholesterol, the triglycerides are also transported in plasma bound to apolipoproteins due to their poor solubility.
- Lipoproteins are synthesized in the liver or intestines and transport fat-soluble substances in the blood, such as cholesterol.
- the lipoproteins are classified according to their density; a distinction is made between the five density classes: chylomicrons, very low density lipoproteines (VDL), low density lipoproteines (LDL) and high density lipoproteines (HDL).
- VDL very low density lipoproteines
- LDL low density lipoproteines
- HDL high density lipoproteines
- the chylomicrons whose physiological concentration in the fasting serum is very low in contrast to that of the other lipoproteins, are transport vehicles for exogenous glycerides.
- the physiological distribution of the other lipoproteins is as follows: VLDL 0%, LDL 70% and HDL 20%.
- VLDL are the forerunners of LDL and vehicles for the transport of endogenous glycerides.
- LDL are created by the hydrolysis of the VLDL.
- LDL and HDL are both regulators of cellular cholesterol homeostasis, with HDL also regulating lipolysis (cleavage of triglycerides into glycerin and free fatty acids).
- LDL have a diameter of approx. 20 nm.
- HDL are the smallest (7-10 nm) and protein richest lipoproteins.
- oxidized LDL (oxLDL) can also be detected in the blood serum.
- oxLDL interacts with endogenous plasma proteins, especially glycoproteins, through specific ligands and forms oxLDL-glycoprotein complexes.
- Apolipoproteins are a component of the lipoproteins and, together with polar lipids, surround the lipoprotein core made of hydrophobic lipids as a kind of outer shell. With the exception of LDL, which only contains apoprotein B, the individual lipoprotein classes have several structurally different ones Apolipoprotein classes.
- Cholesterol is mainly transported by the two lipoprotein classes LDL and HDL. LDL are primarily responsible for the
- LDL cholesterol increases in connection with HDL cholesterol reductions represent the most pronounced risk increase for arteriosclerosis.
- LDL whose particles contribute significantly to the formation of atherosclerotic plaques, and HDL therefore play an opposite role in pathogenesis.
- the quotients of total cholesterol / HDL cholesterol and in particular LDL cholesterol / HDL cholesterol are decisive for assessing the risk of infarction.
- oxLDL-glycoprotein complexes To play role.
- concentration of oxLDL-glycoprotein complexes is higher in patients with chronic kidney failure and diabetes than in healthy patients.
- LDL is removed from the bloodstream by the liver and macrophages.
- Macrophages are cells of the foreign body defense system that are capable of phagocytosis of larger particles.
- the "scavenger pathway” is a well-known model for explaining the uptake of particles by cells (phagocytosis).
- phagocytosis The uptake of solid particles (tissue debris, foreign bodies, bacteria or LDL plaques) into the cell interior of phagocytes with subsequent intracellular degradation takes place by phagocytosis.
- the cells capable of phagocytosis are also referred to as phagocytes and consist mainly of tissue macrophages and mobile ones
- phagocytosis after the particles have been attached to the cell membranes of the phagocytes, by binding to membrane-bound Fc and complement receptors, contractile structures within the cytoplasm are activated. Local indentations of the cell membranes result in the particles being enclosed in cytoplasmic vacuoles.
- the so-called clearing (scavenger) phagocytes can be found in the lymph nodes in and along the fiber strands belonging to the medulla (marrow). During the lymphatic passage from the afferent to the efferent end of the lymph node, particulate antigens are removed by the cells capable of phagocytosis.
- Cells such as polymorphonuclear leukocytes and macrophages, are enlarged by the attachment of immunoglobulins (Ig) to the surface of bacteria (and other antigens). It is believed that the increased adherence is caused by the attachment of the Fc portion of the immunoglobulin to the Fc receptors of the phagocytes.
- Ig immunoglobulins
- the complex of antigen and antibodies is absorbed and digested more quickly by the phagocytic cells.
- the coating of the antibody surface with immunoglobulins is also referred to as opsonin-related (Fc) adherence and plays an important role in immune defense.
- the antibodies that bind to the surface of bacterial cells are able to fix certain components of the extracellular fluids.
- these components are referred to as "complement”.
- Animal experiments have shown that the phagocytosis of cells coated with antibodies is delayed in those animals with a complement deficiency. It therefore suggests that there is a synergism between antibodies and complement during opsonization. Description of the invention
- the effect of the drugs known from the prior art for lowering LDL cholesterol is based on the inhibition of the key enzyme in cholesterol synthesis (CSE).
- the cholesterol synthesis inhibitor is, for example, one under the trade name
- CSE inhibitors in general are significant and include gastrointestinal disorders, sleep disorders, dizziness, visual disturbances, allergic reactions and hair loss. Only in the experimental stage, and only in the case of severe familial hypercholesterolemia, is an approach of somatic gene therapy which involves the transfer of the gene for the LDL receptor to autologous liver cells.
- a substance that is largely free of side effects and acts as a fat reducer is not yet on the market.
- antibodies which induce an increased intracellular accumulation of lipoproteins have so far not been known.
- the damaging role of lipoproteins in kidney diseases lipid-induced damage to cells of the glomerulus filtration apparatus of the kidney
- the object of the invention is to generate a new substance or a new class of substances for the manufacture of a medicament for reducing LDL cholesterol or oxLDL cholesterol in humans and animals with the advantageous aim of reducing the risk of infarction.
- a polypeptide is proposed, the amino acid sequence of which is essentially identical to the amino acid sequence of SEQ ID NO: 1 and / or SEQ ID NO: 3, and the low density lipoproteins (LDL) and / or oxidized LDL ( oxLDL), in particular LDL cholesterol and / or oxidized LDL cholesterol (oxLDL cholesterol) binds.
- the essence of the invention consists in the surprising observation that a purified polypeptide, the sequence of which corresponds in whole or in part to the light (V L ) or heavy chain (VH) of a human monoclonal antibody (SAM-6.10), reduces the low density lipoproteins (LDL) and / or oxLDLs causes.
- V L light
- VH heavy chain
- SAM-6.10 human monoclonal antibody
- the antibody according to the invention comprises the groups V L , V H , Fv, FC, Fab, Fab ', F (ab') 2 designated according to the common nomenclature for describing antibodies.
- the groups mentioned are also referred to as fragments. It is entirely possible that a single fragment is the cause of the fat-lowering effect of the polypeptide according to the invention.
- it is a human monoclonal antibody.
- a “functional fragment” in the sense of the invention is a polypeptide that has at least one of the biological activities that also the entire polypeptide has.
- the specific binding of the antibody can be accomplished, for example, by only one CD region, even though there are a total of 3 CD regions.
- the specific binding of the antibody to an antigen can be used, for example, to induce apoptosis or to inhibit the
- the biological activity of a functional fragment can be measured by various methods known to the person skilled in the art
- the ELISA method is the method for measuring the interaction between antibodies and LDL, in particular LDL cholesterol.
- CDRs complementartity-determining regions of the polypeptide sequence contain the amino acid sequence which is essentially identical to the amino acid sequence Ser-Gly-Asp-Lys-Leu-Gly-Asp-Lys-Tyr-Ala-Cys (CDR1), GIn -Asp-Ser-Lys-Arg-Pro-Ser (CDR2) and Gln-Ala-TrpAsp-Ser-Ser-lle-Val-Val (CDR3) of SEQ ID NO 1 of the light chain variable region (V L ); see also Figure 2.
- CDRs complementarity-determining regions of the peptide sequence contain amino acid sequences which are essentially identical to Ser-Tyr-Ala-Met-His (CDR1), Val-Ile-Ser-Tyr-Asp-Gly-Ser-Asn-Lys- Tyr-Tyr-Ala-Asp-Ser-Val-Lys-Gly (CDR2) and Asp-Arg-Leu-Ala-Val-Ala-Gly-Lys-Thr-Phe-Asp-Tyr (CDR3) of SEQ ID NO 3 the variable region light chain (VH); see also Figure 4.
- a “substantially identical” is a polypeptide or a nucleic acid sequence that is at least 75%, 80%, 85%, or 90% with the amino acid sequence given as a reference (SEQ ID NO:
- polypeptides In a further development of the polypeptide or the nucleic acid sequence, at least 95%, 98%, 99% or 100% identity can be demonstrated in comparison to the references given.
- the length of the comparison section will generally be at least 5, 10, 15 or desirably at least 20 or 25 consecutive amino acids.
- polypeptide according to the invention can be generated by a process which is known under the name Hypridoma technique (Köhler, Millstein,
- kidney diseases in particular glomerulonecrosis (glomerulosclerosis).
- the polypeptide according to the invention is preferably used in the purified form for the production of the medicament, all methods known to the person skilled in the art (e.g. affinity chromatography, gel filtration) being suitable for purification.
- all methods known to the person skilled in the art e.g. affinity chromatography, gel filtration
- the focus is on the fat-lowering effect, in particular the selective lowering of LDL or LDL cholesterol. Due to the property of LDL and / or oxLDL to bind more than
- HDL causes the polypeptide according to the invention to have a low value for the quotient from LDL / HDL and / or oxLDL / HDL and thus minimizes the risk of infarction.
- auxiliaries and excipients for the production of a pharmaceutical are known to the person skilled in the art and can be produced according to common practice (see Remington: The Science and Practice of Pharmacy (20 th ed.), Ed. AR Gennaro, Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JC Boylan, 1988-1999, Marcel Dekker, New York).
- lymphocytes are fused and cultivated with the heteromyeloma HAB-1 (Faller et al., 1990) according to the standard protocol. Briefly summarized, lymphocytes are fused with HAB-1 cells using PEG. The trioms are sown on four 24-hole plates. The average growth frequency is 80-90%, 50% of the growing clones secrete immunoglobulins.
- the secreted human monoclonal antibodies are tested for the first time in an ELISA to determine the isotype. The human monoclonal antibodies can subsequently be analyzed immunohistochemically, genetically, biochemically and molecular biologically.
- RPMI 1640 (company PAA) without additives RPMI 1640 with HAT addition (HAT supplement, company PAA) as well as 10% FCS, 1% glutamine and 1% penicillin / streptomycin HAB-1 (fusion partner) twice with RPMI without additives wash centrifuge thaw frozen lymphocytes (from spleen, lymph nodes or blood) for 5 min at 1500 rpm and wash twice with RPMI without additives, likewise centrifuge both pellets each in 10 ml RPMI without additive and count in a ratio of 1 in the Neubauer counting chamber : 2 - 1: 3, Hab-1 to lymphocytes, fuse the cell pellets together after the second washing process, mix and centrifuge for 8 min at 1500 rpm, the PEG (Polyethylene Glycol 1500, Röche), previously warmed at 37 ° C, carefully Let it drip onto the pellet while rotating the 50 ml tube gently, then resuspend gently and then exactly 90 seconds. then rotate in a water bath at 37 ° C,
- HAT hypoxanthine, aminopterin, thymidine
- the hybridoma cells producing the IgM antibody SAM-6.10 were used for this purpose in a special serum-free cell culture medium (AIMV medium, Gibco) and the IgM content in the culture supernatant was determined nephelometrically.
- AIMV medium serum-free cell culture medium
- the culture supernatant was adjusted to a pH of 5.9 and the solution was filtered.
- a special cation column HiTrap TM SP FF column, 5 ml, Amersham Bioscience was used for binding. The column was equilibrated with filtered buffer A (20 mM phosphate buffer, pH 5.9) at the beginning of the purification.
- the column was washed with buffer A for 20 min and a flow rate of 2 ml / min to a baseline constant in order to remove all unbound proteins.
- the antibody bound to the column was then eluted by admixing buffer B (20 mM phosphate buffer, 1 M NaCl, pH 8.0) and collected in fractions.
- the content of SAM-6.10 antibody (IgM) in the individual fractions was determined nephelometrically below and the purity and
- the measurement is carried out using the Oxidized LDL ELISA test kit from Mercodia, Uppsala, Sweden.
- Test principle The ELISA for oxidized LDL is an enzymatic, bilateral immunoassay with a solid phase. It is based on the direct sandwich technique, in which two monoclonal antibodies against a specific antigen are aligned on oxidized apolipoprotein B. During the incubation, oxidized LDL in the sample reacts with anti-oxidized LDL antibodies, which are bound to the wells in the microtiter plate.
- a peroxidase-conjugated anti-human apolipoprotein B antibody recognizes the oxidized LDL bound to the solid phase. After a second incubation and a washing process which removes the antibodies labeled with the unbound enzyme, the conjugate is reacted with
- TMB 5'-tetramethylbenzidine
- Trypsin / EDTA detached from the bottom of the culture bottles.
- the reaction was stopped with 10 ml of RPMI-1640 medium (+ supplements) and the cells were pelleted at 1000 ⁇ g for 5 min.
- the cells were washed twice with PBS, taken up in FACS buffer (PBS, 0.01% Na azide) and placed on ice for 30 min to reconstitute the cell membranes.
- the cells were then adjusted to a density of 1 ⁇ 10 6 cells / ml and each 200 ⁇ l of the cell suspensions were transferred to FACS reaction vessels, so that there was a cell count of 2 ⁇ 10 5 cells per tube.
- the cells were pelleted for 5 min at 4 ° C.
- the pellets resuspended and incubated with the primary antibodies on ice for 15 min.
- 100 ⁇ g / ml SAM-6 antibodies in FACS buffer (total volume 200 ⁇ l) or 100 mg / ml LDL were used as primary antibodies.
- 100 ⁇ g / ml Chrompure human IgM acted as an isotype control.
- the cells were pre-incubated with LDL for 30 min and then for 15 min
- F ITC-coupled secondary antibody (Rabbit anti human IgM, F ITC-coupled, 1:50 in FACS buffer for SAM-6 or Chrompure human IgM). After another wash, the cells were taken up in 200 ⁇ l cold FACS buffer and kept on ice until protected from light until measurement. The measurement was carried out using a
- Experiment 1 500 ⁇ g of purified SAM-6.10 antibody was injected intra peritoneally. The LDL concentration in the blood serum was checked
- Experiment 3 1 mg of purified SAM-6.10 antibodies were injected intraperitoneally. The LDL concentration in the blood serum was measured after 14 days.
- Control A normal values for control mouse
- Control B normal values for control mouse
- the serum concentration of LDL is significantly reduced in the mice treated with SAM-6.10 (see FIGS. 10 and 11).
- liver, lungs, heart, spleen, small intestine, colon, kidneys, stomach and brain showed no morphological changes.
- organs were immunohistochemically examined for possible lipid deposits. examined. Staining with Sudan III showed no lipid storage in any organ.
- the organs of the killed mice were fixed in formalin and embedded in paraffin.
- the dyeing was carried out using the Sudan III dye according to the following protocol:
- the adherent macrophages are grown on object supports and then treated with the appropriate agents.
- the staining is carried out as follows: Fixation of the cells in 60% isopropanol (6 min) • Incubate 20 minutes with Sudan III
- the measurement of the different lipids in the blood serum was carried out automatically with the MODULAR D P800 device (Röche).
- the LDL cholesterol value was determined using an enzymatic, colorimetric method (CHOD / PAP) without sample pretreatment.
- HDL, VLDL and chylomicrons are specifically hydrolyzed by a detergent 1.
- the cholesterol released in these lipoproteins reacts immediately through the enzymatic action of cholesterol esterase (CE) and cholesterol oxidase (CHOD) and hydrogen peroxide is formed.
- CE cholesterol esterase
- CHOD cholesterol oxidase
- the LDL particles remain intact step by step.
- the reaction of LDL cholesterol is triggered by adding detergent 2 and the coupling substance N, N-bis (4-sulfobutyl) -m-toluidine (DSBmT).
- the second detergent releases cholesterol in the LDL particles.
- a dye is formed in the enzymatic reaction in the presence of the coupling substance.
- the intensity of the red quinonimine dye formed is directly proportional to the LDL cholesterol concentration. It is determined by measuring the increase in absorbance at 552 nm.
- ELISA LDL / HDL
- LDL Lipoprotein Low Density tyfrom Human Plasma, Sigma, 10 ⁇ g / ml in PBS
- HDL Human HDL, Chemicon, 10 ⁇ g / ml in PBS
- Figure 1 shows the amino acid sequences (SEQ ID NO 1) of the variable region of the light chain (VL).
- Figure 2 shows the nucleotide acid sequence (SEQ ID NO 2) of the light chain variable region (VL).
- the complementarity-determining regions (CDR) are marked by dashes and are essentially identical to nucleotides 67-99 (CDR1), 145-165
- Figure 3 shows the amino acid sequences (SEQ ID NO 3) of the heavy chain variable region (VH).
- Figure 4 shows the nucleotide acid sequence (SEQ ID NO 4) of the heavy chain variable region (VH).
- the complementarity-determining regions (CDR) are identified by dashes and are essentially identical to nucleotides 91-105 (CDR1), 148-198 (CDR2) and 295-330 (CDR3) from SEQ ID NO 4.
- FIG. 5 shows the measurement of oxLDL as a function of the incubation period with a copper sulfate solution.
- LDL Sigma, Taufkirchen, Germany
- ELISA determined the amount of oxi- dated LDL determined. It can be clearly seen that the amount of oxidized LDL increases with increasing incubation time, and even that LDL fraction that has not been treated with copper ions is partly already in oxidized form. After 15 hours of incubation, the proportion of oxidized LDL has approximately doubled.
- FIG. 6 shows the detection of the binding of SAM-6 to oxLDL.
- the ELISA plate was pre-coated with differently oxidized LDL fractions before the first antibody
- Figure 7 shows the result of a FACS analysis.
- the cells used for this are cells from the mouse macrophage cell line P388D1 (IL-1) (DSMZ Accession No. ACC 288).
- Figure 7A shows the binding of LDL to macrophages.
- FIG. 7B shows that the human monoclonal antibody SAM-6 also binds to macrophages.
- FIG. 7C shows that the macrophages have ⁇ -receptors.
- FIG. 7D With the right shift of the signal in FIG. 7D it is shown that a simultaneous incubation of LDL and SAM-6 brings about an increased binding of SAM-6 to the cells.
- FIG. 8 shows cells which were incubated for 48 hours with either SAM-6 or an IgM control antibody and then subjected to Sudan III staining.
- SAM-6 show a clear deposition of neutral fats due to the red color.
- Those incubated with the control antibody show a clear deposition of neutral fats due to the red color.
- FIG. 9 For the stains shown in FIG. 9, the macrophages were cultivated for 24 hours both with and without the addition of FCS. Thereafter, only LDL or only SAM-6 or LDL and SAM-6 were added together for a further 24 hours. A Sudan III staining was then carried out.
- the left column of the image with FIGS. 9A, 9C and 9E show cells that were cultivated without the addition of FCS.
- the right-hand image column with FIGS. 9B, 9D and 9F show cells which were cultivated with the addition of FCS.
- FIGS. 9A and 9B show that both macrophages which have been cultivated without FCS and macrophages which have grown in the presence of FCS show a basal storage of neutral fats.
- FIG. 9C shows that when
- FIGS. 9E and 9F show that with a co-incubation of SAM-6 and LDL, the intracellular lipid accumulation increases markedly both in macrophages which have been cultured with FCS and in those which have grown without the addition of FCS.
- FIG. 10 shows the influence of the antibody SAM-6 on LDL values in vivo.
- the mice were given 1 mg of purified SAM-6 antibody per or in the control experiment 1 mg human Chrompure IgM (isotype control) ip injected.
- the LDL concentration in the blood was measured after 24 and 48 hours. A clear reduction in serum LDL can be observed after 24 and 48 hours in mice treated with SAM-6.
- FIG. 11 also demonstrates the in vivo effect of SAM-6.10, an indirect method using the Friedewald formula being used to calculate the values.
- the amount of LDL is calculated from the difference between total cholesterol, HDL and triglycerides. According to this type of test evaluation, the reduction in the concentration of LDL in the serum after SAM-6.10 treatment would be even greater. However, this indirect measurement method must be regarded as less precise than the method used in FIG. 10.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Obesity (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Child & Adolescent Psychology (AREA)
- Urology & Nephrology (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Description
Claims
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE112004002660T DE112004002660D2 (de) | 2003-11-14 | 2004-11-12 | Humaner monoklonaler Antikörper mit fettsenkender Wirkung |
EP04802719.7A EP1682580B1 (de) | 2003-11-14 | 2004-11-12 | Humaner monoklonaler antikörper mit fettsenkender wirkung |
ES04802719T ES2424639T3 (es) | 2003-11-14 | 2004-11-12 | Anticuerpo monoclonal humano con efecto reductor de la grasa |
US10/578,856 US8124080B2 (en) | 2003-11-14 | 2004-11-12 | Human monoclonal antibody having fat-reducing effect |
JP2006543353A JP4980068B2 (ja) | 2003-11-14 | 2004-11-12 | 脂肪を減少する効果を有するヒト単クローン抗体 |
US13/406,191 US9273125B2 (en) | 2003-11-14 | 2012-02-27 | Human monoclonal antibody having fat-reducing effect |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE10353175A DE10353175A1 (de) | 2003-11-14 | 2003-11-14 | Humaner monoklonaler Antikörper mit fettsenkender Wirkung |
DE10353175.0 | 2003-11-14 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10/578,856 A-371-Of-International US8124080B2 (en) | 2003-11-14 | 2004-11-12 | Human monoclonal antibody having fat-reducing effect |
US13/406,191 Continuation US9273125B2 (en) | 2003-11-14 | 2012-02-27 | Human monoclonal antibody having fat-reducing effect |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2005049635A2 true WO2005049635A2 (de) | 2005-06-02 |
WO2005049635A3 WO2005049635A3 (de) | 2006-02-02 |
Family
ID=34585082
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/DE2004/002503 WO2005049635A2 (de) | 2003-11-14 | 2004-11-12 | Humaner monoklonaler antikörper mit fettsenkender wirkung |
Country Status (6)
Country | Link |
---|---|
US (2) | US8124080B2 (de) |
EP (2) | EP2374820A3 (de) |
JP (1) | JP4980068B2 (de) |
DE (2) | DE10353175A1 (de) |
ES (1) | ES2424639T3 (de) |
WO (1) | WO2005049635A2 (de) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2431053A1 (de) | 2006-11-27 | 2012-03-21 | Patrys Limited | Neues glykosyliertes Zielpeptid in neoplastischen Zellen |
US8163552B2 (en) | 2003-11-14 | 2012-04-24 | Patrys Limited | Adenocarcinoma specific antibody SAM-6, and uses thereof |
WO2013104918A2 (en) * | 2012-01-13 | 2013-07-18 | Imperial Innovations Limited | Binding molecule |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010088739A1 (en) * | 2009-02-09 | 2010-08-12 | Patrys Limited | Sam-6 variants, target and methods of use |
EP2789629A1 (de) * | 2013-04-11 | 2014-10-15 | Universiteit Maastricht | Verfahren zur Behandlung von lysosomalen Fettspeicherkrankheiten |
JP7403241B2 (ja) * | 2019-06-07 | 2023-12-22 | 小林製薬株式会社 | 内臓脂肪の視認性向上方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003048321A2 (en) | 2001-12-03 | 2003-06-12 | Alexion Pharmaceuticals, Inc. | Hybrid antibodies |
Family Cites Families (49)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5374548A (en) | 1986-05-02 | 1994-12-20 | Genentech, Inc. | Methods and compositions for the attachment of proteins to liposomes using a glycophospholipid anchor |
US5126240A (en) * | 1986-09-29 | 1992-06-30 | Curtiss Linda K | Hybridomas and monoclonal paratopic molecules to apolipoprotein a-i |
ATE121197T1 (de) * | 1988-01-19 | 1995-04-15 | Pasteur Institut | Immunometrisches verfahren zum nachweis von lipoprotein-teilchen, anti-lp(a) monoklonale antikörper und ihre anwendung zur diagnose von atherosklerose. |
DE4107154A1 (de) | 1990-10-11 | 1992-04-16 | Boehringer Mannheim Gmbh | Monoklonale antikoerper gegen melanom |
AU662311B2 (en) | 1991-02-05 | 1995-08-31 | Novartis Ag | Recombinant antibodies specific for a growth factor receptor |
GB9105292D0 (en) | 1991-03-13 | 1991-04-24 | Sandoz Ltd | Improvements in or relating to organic compounds |
US5639641A (en) | 1992-09-09 | 1997-06-17 | Immunogen Inc. | Resurfacing of rodent antibodies |
US5622701A (en) | 1994-06-14 | 1997-04-22 | Protein Design Labs, Inc. | Cross-reacting monoclonal antibodies specific for E- and P-selectin |
US5639863A (en) | 1994-06-21 | 1997-06-17 | Dan; Michael D. | Human monoclonal antibodies specific to cell cycle independent glioma surface antigen |
US6429289B1 (en) * | 1994-06-23 | 2002-08-06 | Massachusetts Institute Of Technology | Class BI and CI scavenger receptors |
GB9424449D0 (en) | 1994-12-02 | 1995-01-18 | Wellcome Found | Antibodies |
US6140470A (en) | 1995-06-30 | 2000-10-31 | Yale University | Human monoclonal anti-tumor antibodies |
GB9601081D0 (en) | 1995-10-06 | 1996-03-20 | Cambridge Antibody Tech | Specific binding members for human transforming growth factor beta;materials and methods |
AU4982097A (en) | 1996-10-18 | 1998-05-15 | Board Of Regents, The University Of Texas System | Anti-erbb2 antibodies |
EP0927757A1 (de) | 1997-12-03 | 1999-07-07 | Leadd B.V. | Verfahren und Mitteln um Apoptose zu induzieren durch Interferenz mit Bip-ähnliche Proteine |
CA2319089A1 (en) | 1998-04-09 | 1999-10-21 | Genset S.A. | 5' ests and encoded human proteins |
DK1087993T3 (da) | 1998-06-18 | 2008-07-14 | Imed Ab | Fas-peptider og antistoffer til at modulere apoptose |
DE69901569T2 (de) | 1998-08-28 | 2002-12-19 | Genentech Inc | Humane antikörper gegen faktor ix/ixa |
GB9822115D0 (en) | 1998-10-09 | 1998-12-02 | King S College London | Treatment of inflammatory disease |
PT1141019E (pt) | 1998-12-22 | 2004-09-30 | Heinz Peter Vollmers | Fragmento da glicoproteina cd55/daf para a obtencao de medicamentos antitumorais com elevada eficacia assim como processo para a sua utilizacao |
CN101066997B (zh) | 1999-03-25 | 2013-03-27 | 艾博特股份有限两合公司 | 结合人il-12的人抗体及其生产方法 |
US7049132B1 (en) | 1999-06-28 | 2006-05-23 | University Of Southern California | Stress-responsive induction of a therapeutic agent and methods of use |
US6677442B1 (en) | 1999-10-29 | 2004-01-13 | University Of Kentucky Research Foundation | Nucleic acid encoding human REV1 protein |
MXPA02008144A (es) | 2000-02-23 | 2002-11-29 | Amgen Inc | Agentes de enlace selectivos antagonistas de la proteina de enlace de osteoprotegerina. |
TWI318983B (en) | 2000-05-02 | 2010-01-01 | Uab Research Foundation | An antibody selective for a tumor necrosis factor-related apoptosis-inducing ligand receptor and uses thereof |
ES2609016T3 (es) | 2000-06-16 | 2017-04-18 | Human Genome Sciences, Inc. | Anticuerpos que se unen inmunoespecíficamente a BLyS |
UA81743C2 (uk) | 2000-08-07 | 2008-02-11 | Центокор, Инк. | МОНОКЛОНАЛЬНЕ АНТИТІЛО ЛЮДИНИ, ЩО СПЕЦИФІЧНО ЗВ'ЯЗУЄТЬСЯ З ФАКТОРОМ НЕКРОЗУ ПУХЛИН АЛЬФА (ФНПα), ФАРМАЦЕВТИЧНА КОМПОЗИЦІЯ, ЩО ЙОГО МІСТИТЬ, ТА СПОСІБ ЛІКУВАННЯ РЕВМАТОЇДНОГО АРТРИТУ |
US20030105003A1 (en) | 2001-04-05 | 2003-06-05 | Jan Nilsson | Peptide-based immunization therapy for treatment of atherosclerosis and development of peptide-based assay for determination of immune responses against oxidized low density lipoprotein |
CA2443862A1 (en) | 2001-04-17 | 2002-10-24 | Peizhi Luo | Structure-based construction of human antibody library |
CN1558914A (zh) | 2001-07-24 | 2004-12-29 | ��˹�������¡�����-��÷���� | 受体、其应用以及小鼠抗体 |
US7671010B2 (en) | 2002-08-30 | 2010-03-02 | The Board Of Regents Of The University Of Texas System | Compositions and methods of use of targeting peptides for diagnosis and therapy of human cancer |
US20040048243A1 (en) | 2001-09-07 | 2004-03-11 | Wadih Arap | Methods and compositions for in vitro targeting |
DE10210427A1 (de) | 2002-03-09 | 2003-10-09 | Hans Konrad Mueller-Hermelink | Humaner monoklonaler Antikörper |
EP1572079A4 (de) * | 2002-03-29 | 2006-09-06 | Centocor Inc | Säugetier-cdr-mimetikörper, zusammensetzungen, verfahren und anwendungszwecke |
EP2322559A3 (de) | 2002-07-04 | 2011-08-03 | Patrys Limited | Neoplasma-spezifische Antikörper und deren Verwendungen |
DE10230516A1 (de) | 2002-07-06 | 2004-01-15 | Müller-Hermelink, Hans Konrad, Prof. Dr. | Humaner monoklonaler Antikörper |
AU2002364501A1 (en) | 2002-08-30 | 2004-03-19 | Board Of Regents, The University Of Texas System | Compositions and methods of use of targeting peptides for diagnosis and therapy of human cancer |
SE0302312D0 (sv) | 2002-10-04 | 2003-08-27 | Forskarpatent I Syd Ab | Peptide-based passive immunization therapy for treatment of atherosclerosis |
DE10311248A1 (de) | 2003-03-14 | 2004-09-30 | Müller-Hermelink, Hans Konrad, Prof. Dr. | Humaner monoklonaler Antikörper |
WO2005001052A2 (en) | 2003-06-06 | 2005-01-06 | University Of Massachusetts | Modulation of apoptosis |
AU2004288166B2 (en) | 2003-10-27 | 2008-12-18 | University Of Southern California | Methods and compositions for modulating apoptosis |
NZ577824A (en) | 2003-11-12 | 2010-10-29 | Patrys Ltd | Methods of identifying neoplasm-specific antibodies and uses thereof |
EP1531162A1 (de) | 2003-11-14 | 2005-05-18 | Heinz Vollmers | Antikörper SAM-6, der spezifisch von Adenocarzinom ist, und dessen Verwendungen |
WO2005065418A2 (en) | 2003-12-31 | 2005-07-21 | Board Of Regents, The University Of Texas System | Compositions and methods of use of targeting peptides for diagnosis and therapy |
GB0404936D0 (en) | 2004-03-04 | 2004-04-07 | Univ London | Screen |
DE102004015179A1 (de) | 2004-03-25 | 2005-10-13 | Oncomab Gmbh | Antigen des PM-2 Antikörpers und dessen Verwendung |
ES2476798T3 (es) | 2006-11-27 | 2014-07-15 | Patrys Limited | Nueva diana de p�ptido glucosilado en células neopl�sicas |
WO2009104100A2 (en) * | 2008-02-19 | 2009-08-27 | Patrys Limited | Antibody combinations, and methods of making and using same |
WO2010088739A1 (en) | 2009-02-09 | 2010-08-12 | Patrys Limited | Sam-6 variants, target and methods of use |
-
2003
- 2003-11-14 DE DE10353175A patent/DE10353175A1/de not_active Withdrawn
-
2004
- 2004-11-12 EP EP10194202A patent/EP2374820A3/de not_active Withdrawn
- 2004-11-12 ES ES04802719T patent/ES2424639T3/es active Active
- 2004-11-12 DE DE112004002660T patent/DE112004002660D2/de not_active Expired - Fee Related
- 2004-11-12 WO PCT/DE2004/002503 patent/WO2005049635A2/de active Application Filing
- 2004-11-12 US US10/578,856 patent/US8124080B2/en not_active Expired - Fee Related
- 2004-11-12 EP EP04802719.7A patent/EP1682580B1/de not_active Not-in-force
- 2004-11-12 JP JP2006543353A patent/JP4980068B2/ja not_active Expired - Fee Related
-
2012
- 2012-02-27 US US13/406,191 patent/US9273125B2/en not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003048321A2 (en) | 2001-12-03 | 2003-06-12 | Alexion Pharmaceuticals, Inc. | Hybrid antibodies |
Non-Patent Citations (1)
Title |
---|
KÖHLER, MILLSTEIN, NATURE, vol. 256, 1975, pages 495 |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8163552B2 (en) | 2003-11-14 | 2012-04-24 | Patrys Limited | Adenocarcinoma specific antibody SAM-6, and uses thereof |
US8741296B2 (en) | 2003-11-14 | 2014-06-03 | Patrys Limited | Adenocarcinoma specific antibody SAM-6, and uses thereof |
EP2431053A1 (de) | 2006-11-27 | 2012-03-21 | Patrys Limited | Neues glykosyliertes Zielpeptid in neoplastischen Zellen |
WO2013104918A2 (en) * | 2012-01-13 | 2013-07-18 | Imperial Innovations Limited | Binding molecule |
WO2013104918A3 (en) * | 2012-01-13 | 2013-08-29 | Imperial Innovations Limited | Binding molecule |
Also Published As
Publication number | Publication date |
---|---|
DE10353175A1 (de) | 2005-06-16 |
JP2007537995A (ja) | 2007-12-27 |
US8124080B2 (en) | 2012-02-28 |
US9273125B2 (en) | 2016-03-01 |
US20080045701A1 (en) | 2008-02-21 |
EP2374820A2 (de) | 2011-10-12 |
WO2005049635A3 (de) | 2006-02-02 |
JP4980068B2 (ja) | 2012-07-18 |
EP1682580B1 (de) | 2013-05-15 |
EP2374820A3 (de) | 2012-01-11 |
ES2424639T3 (es) | 2013-10-07 |
EP1682580A2 (de) | 2006-07-26 |
US20130072664A1 (en) | 2013-03-21 |
DE112004002660D2 (de) | 2006-10-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
DE60226036T3 (de) | ANTIKÖRPER, DER DAS GM1-GANGLIOSID-GEBUNDENE AMYLOID-b-PROTEIN ERKENNT, UND DNA, DIE FÜR DIESEN ANTIKÖRPER CODIERT | |
DE3439345C2 (de) | Verwendung von hyperimmuner Milch zur Behandlung von Herzgefäßerkrankungen bei Tier und Mensch | |
DE69925909T2 (de) | T-zell inhibierende rezeptorzusammensetzungen sowie deren verwendung | |
DE69735294T2 (de) | Spezifische monoklonale antikörper für die extrazelluläre domäne von protasta-spezifischem membranantigen | |
DE69333209T2 (de) | Angiogenese-inhibierende antikörper | |
DE60018761T2 (de) | Therapeutischer antikörper gegen das muc-1-antigen und verfahren zu dessen verwendung | |
EP0480440B1 (de) | Monoklonale Antikörper gegen Melanom | |
DE69433563T2 (de) | Gegen das ca55.1 antigen gerichtete bindende strukturen | |
DE60223688T2 (de) | Verfahren zur behandlung von multiplem myelom | |
Morace et al. | Renal globotriaosylceramide facilitates tubular albumin absorption and its inhibition protects against acute kidney injury | |
US9273125B2 (en) | Human monoclonal antibody having fat-reducing effect | |
DE69911322T2 (de) | Monoklonaler antikörper der die oligosaccharide sialinsäure n-glycolierte galactose-glucose auf bösartige tumoren erkennt und zusammensetzungen die dieser enthalten | |
DE60113976T2 (de) | Funktionaler nachweis von lipoprotein mit hoher dichte | |
Kikuchi et al. | Severe proteinuria, sustained for 6 months, induces tubular epithelial cell injury and cell infiltration in rats but not progressive interstitial fibrosis | |
EP2129403B1 (de) | Bispezifisches polypeptidisches Konstrukt mit therapeutischem und diagnostischem Potential | |
DE10210427A1 (de) | Humaner monoklonaler Antikörper | |
EP1747011A2 (de) | Mittel und verfahren zur diagnose, prophylaxe und therapie von bindegewebserkrankungen | |
WO2007012449A1 (de) | Frizzled 9 als tumormarker | |
JP2007537995A5 (de) | ||
DE4120213C2 (de) | Monoklonale Antikörper gegen das TNF-bindende Protein I (TNF-BP I) | |
WO1992022666A1 (de) | Monoklonale antikörper gegen das humane tnf-bindende protein i (tnf-bp i) | |
AT502227B1 (de) | Verfahren zur diagnose von tumoren | |
EP0618815B1 (de) | Monoklonale antikörper gegen die typ i-phospholipase a 2? als entzündungshemmendes therapeutikum | |
DE60036452T2 (de) | Monoklonaler anti-uracil antikörper | |
DE4103727A1 (de) | Zur spezifischen erkennung eines antigens des stratum corneum der menschlichen epidermis befaehigte antikoerper, ihre herstellung und ihre anwendung, insbesondere als traeger von arzneimittelwirkstoffen |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A2 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2004802719 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2006543353 Country of ref document: JP |
|
DPEN | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed from 20040101) | ||
WWE | Wipo information: entry into national phase |
Ref document number: 10578856 Country of ref document: US |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1120040026601 Country of ref document: DE |
|
WWP | Wipo information: published in national office |
Ref document number: 2004802719 Country of ref document: EP |
|
REF | Corresponds to |
Ref document number: 112004002660 Country of ref document: DE Date of ref document: 20061005 Kind code of ref document: P |
|
WWE | Wipo information: entry into national phase |
Ref document number: 112004002660 Country of ref document: DE |
|
WWP | Wipo information: published in national office |
Ref document number: 10578856 Country of ref document: US |