WO1981001145A1 - Hydrolytic enzyme-activatible pro-drugs - Google Patents

Hydrolytic enzyme-activatible pro-drugs

Info

Publication number
WO1981001145A1
WO1981001145A1 PCT/US1980/001290 US8001290W WO1981001145A1 WO 1981001145 A1 WO1981001145 A1 WO 1981001145A1 US 8001290 W US8001290 W US 8001290W WO 1981001145 A1 WO1981001145 A1 WO 1981001145A1
Authority
WO
Grant status
Application
Patent type
Prior art keywords
drug
moiety
amino acid
acid residue
pro
Prior art date
Application number
PCT/US1980/001290
Other languages
French (fr)
Inventor
P Carl
J Katzenellenbogen
M Weber
Original Assignee
Univ Illinois
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date

Links

Classifications

    • BPERFORMING OPERATIONS; TRANSPORTING
    • B82NANOTECHNOLOGY
    • B82YSPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/65Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6891Pre-targeting systems involving an antibody for targeting specific cells
    • A61K47/6899Antibody-Directed Enzyme Prodrug Therapy [ADEPT]
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06Esters of carbamic acids
    • C07C271/08Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
    • C07C271/26Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atom of at least one of the carbamate groups bound to a carbon atom of a six-membered aromatic ring
    • C07C271/28Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atom of at least one of the carbamate groups bound to a carbon atom of a six-membered aromatic ring to a carbon atom of a non-condensed six-membered aromatic ring
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D261/00Heterocyclic compounds containing 1,2-oxazole or hydrogenated 1,2-oxazole rings
    • C07D261/02Heterocyclic compounds containing 1,2-oxazole or hydrogenated 1,2-oxazole rings not condensed with other rings
    • C07D261/04Heterocyclic compounds containing 1,2-oxazole or hydrogenated 1,2-oxazole rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575Hormones
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K5/00Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08Tripeptides
    • C07K5/0802Tripeptides with the first amino acid being neutral
    • C07K5/0804Tripeptides with the first amino acid being neutral and aliphatic
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K5/00Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10Tetrapeptides
    • C07K5/1002Tetrapeptides with the first amino acid being neutral
    • C07K5/1005Tetrapeptides with the first amino acid being neutral and aliphatic
    • C07K5/101Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K38/00Medicinal preparations containing peptides

Abstract

Antineoplastic agents are rendered tumor-specific by derivatization with a peptide specifier so as to convert the antineoplastic agent into a pharmacologically inactive pro-drug which is selectively activatible at the tumor site. The peptide specifier has an amino acid residue sequence such that it will be selectively enzymatically cleaved from the antineoplastic agent by tumor-associated fibrinolytic and/or blood-coagulating proteases, such as plasmin and plasminogen activator, so as to effect release of the antineoplastic agent in pharmacologically active form in the vicinity of the tumor. These and other similar hydrolytic enzyme-activatible pro-drugs may be formed with their specifier moiety and their drug moiety covalently linked together through an intermediate self-immolative connector moiety having a molecular structure such that enzymatic cleavage of the bond covalently linking it to the specifier moiety will initiate spontaneous cleavage of the bond covalently linking it to the drug moiety to thereby effect release of the drug in pharmacologically active form.

Description

Description

HYDROLYTIC ENZYME-ACTIVATIBLE PRO-DRUGS

BACKGROUND OF THE INVENTION This invention relates to hydrolytic enzyme- activatible pro-drugs and, in particular, to tumor- specific pro-drugs of antineoplastic agents which are selective substrates for drug-activating enzymatic cleavage by tumor-associated proteases.

One approach to improving the efficiency of drug action and the selectivity of drug delivery is to prepare a reversible derivative of a drug which is itself pharmacologically inactive, but which becomes activated in vivo to liberate the parent drug, typically, but not necessarily, by enzymatic attack. A drug derivative of this type, commonly known as a "pro- drug" or a "latentiated drug", can be tailored to overcome certain undesirable properties of the parent drug, such as, for example, bitterness or tartness, offensive odor, gastric or intestinal upset and irritation, pain on injection, lack of absorption, slow or rapid metabolism, or lack of stability in the bulk state, the dosage form, or in vivo; or it can be designed to be activated selectively at the site of intended action, so that undesired effects can be lessened, The present invention is primarily concerned with pro-drugs of this latter type, which are selectively activatible at the site of intended action, and, in particular, to pro-drugs of antineoplastic agents which are selectively activatible at the tumor site. One aspect of the present invention, however, is more broadly applicable to pro-drugs in general, as will become more readily apparent hereinbelow.

Many of the antineoplastic agents currently being used in cancer chemotherapy rely for their effectiveness on being selectively cytotoxic to rapidly proliferating cells. In addition to malignant cells, however, certain normal cells are also rapidly proliferating, such as, for example, bone marrow and spleen cells. For this reason bone marrow and spleen toxicity are often limiting factors in the effectiveness of such antineoplastic agents in cancer chemotherapy. One approach in trying to overcome this problem is to design a pro-drug of the antineoplastic agent which will be selectively activatible at the tumor-site, for example, by being a selective substrate for drug-activating enzymatic cleavage by a tumor-associated enzyme. In order for a pro-drug of this type to be useful in cancer chemotherapy, there are several criteria which it must meet. First of all, there must be enough of the activating enzyme in the tumor to generate cytotoxic levels of free drug in the vicinity of the tumor. Secondly, there must be means available to minimize activation of the pro-drug at sites distant from the tumor, and to mitigate the effects of such activation if it occurs. This criterion is clearly related to the first one, since it is the relative amount of tumor-associated and extra-tumor enzymatic activity which is critical for selectivity. Thirdly, the pro-drug must be a suitable substrate for the tumor-associated enzyme under physiological conditions and a poor substrate for other enzymes. Fourthly, the pro-drug must be considerably less toxic than the activated drug, i^.e., at least on the order of ten times less active and preferably on the order of a hundred or a thousand times less active. Finally, the activated species must have a reasonably short biological half-life so that the toxic effects of the locally activated drug are limited to the tumor and selectivity is not lost by diffusion of the drug away from the site of activation.

A number of attempts have previously been made to develop tumor-specific pro-drugs of antineoplastic agents activatible by tumor-associated enzymes. However, such previous attempts have met with very limited success primarily due to a failure of such pro-drugs to meet one or more of the five criteria set forth above.

Another consideration in the design of hy drolytic enzyme-activatible pro-drugs, in general, is the problem sometimes posed by the nature of the drug molecule being derivatized. If the drug molecule is large or has pronounced polar or apolar character, steric or electronic factors at the intended cleavage site could interfere with the enzymatic cleavage reaction and thereby prevent the pro-drug from being a suitable substrate for the target enzyme.

It has previously been reported that many animal and human tumors exhibit elevated levels of fibrinolytic and blood-coagulating enzyme activity and, in particular, elevated levels of the fibrinolytic enzymes, plasmin and plasminogen activator. Both plasmin and plasminogen activator are proteases with trypsin-like specificity in the sense that they both cleave next to basic amino acids. Substantial information exists concerning the specificity of plasmin and plasminogen activator, based on the use of artificial substrates as well as analysis of the peptide bonds cleaved in the natural substrate. Plasminogen activator shows considerable substrate specificity towards its natural substrate, plasminogen, in which a single Arg-Val bond is cleaved in converting plasminogen to plasmin. Plasmin is often regarded as a rather unspecific, trypsin-like protease. However, it cleaves a limited number of bonds in dissolving a fibrin clot. Examination of the plasmin cleavage sites in its natural substrate, fibrin, reveal that eleven of the fifteen earliest cleavages are at lysine residues, and in fifteen of the twenty earliest cleavages (including all of the earliest nine cleavages) a hydropho amino acid precedes the lysine or arginine. Hence, the implication is that plasmin is selective for lysine residues preceded by a hydrophobic amino acid residue.

The elevated levels of fibrinolytic and blood- coagulating enzymes found in many tumor cells and the substrate specificity of these proteases have not previously been exploited in the design of tumor- specific pro-drugs of antineoplastic agents. While it is true that various normal cells and tissues, including the lung, kidney, squamous epithelium and activated macrophages, also exhibit elevated levels of these proteases, such normal cells by and large are not rapidly proliferating, and thus should not be highly sensitive to the cytotoxic effects of a DNA synthesis inhibitor released in their vicinity. On the other hand, at least two major sites of high normal cell proliferation, i.e., the bone marrow and spleen, have been reported to be low in fibrinolytic and blood-coagulating enzyme activity. Hence, the specifi combination of rapidly proliferating cells exhibiting high levels of fibrinolytic and blood-coagulating enzyme activity appears to be a characteristic possessed by a great many tumor cells but generally not possessed by normal cells. SUMMARY OF THE INVENTION It is, accordingly, a primary object of the present invention to provide a pro-drug of an antineoplastic agent which is selectively activatible at the site of the tumor.

Another object of the invention is to provide a tumor-specific pro-drug of an antineoplastic agent in accordance with the preceding object, which is a highly selective substrate for drug-activating enzymatic cleavage by one or more tumor-associated hydrolytic enzymes.

A further object of the invention is to provide a tumor-specific pro-drug of an antineoplastic agent in accordance with the preceding objects, wherein the activating enzyme is one which is present in the tumor in sufficient amounts to generate cytotoxic levels of free drug in the vicinity of the tumor.

Still another object of the invention is to provide a tumor-specific pro-drug of an antineoplastic agent in accordance with the preceding objects, wherein the activating enzyme is one whose presence at sites distant from the tumor is insufficient to generate cytotoxic levels of free drug in the vicinity of such distant sites.

A still further object of the present invention is to provide a tumor-specific pro-drug of an antineoplastic agent in accordance with the preceding objects, which is considerably less toxic than the activated drug.

Yet another object of the present invention is to provide a tumor-specific pro-drug of an antineoplastic agent in accordance with the preceding objects, wherein the activated drug has a reasonably short biological half-life so that the cytotoxic ef- fects of the locally activated drug are limited to the tumor and selectivity is not lost by diffusion of the drug away from the site of activation.

A yet further object of the present invention is to provide hydrolytic enzyme-activatible pro- drugs, including those of the type set forth in the preceding objects, which include connector means for spacing the drug-activating enzymatic cleavage site sufficiently far away from the drug molecule so as to prevent steric and/or electronic interference with the enzymatic cleavage reaction, which connector means does not in itself prevent release of the free drug in pharmacologically active form following the enzymatic cleavage reaction. The above and other objects are achieved in accordance with the present invention by derivatizing an antineoplastic agent with a peptide specifier at a reactive site appropriate for inhibiting the pharmacological activity of the antineoplastic agent, to thereby convert the antineoplastic agent into a pharmacologically inactive peptidyl derivative pro- drug. The peptide specifier has an amino acid residue sequence specifically tailored so as to render the peptidyl derivative a selective substrate for drug-activating enzymatic cleavage by one or more tumor-associated fibrinolytic and/or blood-coagulating proteases, such as plasmin and plasminogen activator. The enzymatic cleavage reaction will remove the peptide specifier moiety from the pro-drug and effect release of the antineoplastic agent in pharmacologically active form selectively at the tumor site.

In those instances where the drug molecule is large and/or has pronounced polar or apolar character, steric and/or electronic interference of the enzymatic cleavage reaction is avoided in accordance with the present invention by forming the peptidyl derivative pro-drug with its peptide specifier moiety and its antineoplastic agent moiety covalently linked together through an intermediate self-immolative connector moiety having a molecular structure such that enzymatic cleavage of the bond covalently linking it to the peptide specifier moiety will initiate spontaneous cleavage of the bond covalently linking it to the antineoplastic agent moiety to thereby effect release of the antineoplastic agent in pharmacologically active form. The intermediate self- immolative connector aspect of the present invention is not limited in its application to protease-activatible pro-drugs of antineoplastic agents, but is equally applicable to a variety of other types of hydrolytic enzyme-activatible pro-drugs wherein steric and/or electronic hindrance by the drug molecule might otherwise interfere with the drug-activating enzymatic cleavage reaction. Moreover, the self-immolative connector aspect of the present invention may also be used to impart to the pro-drugs greater stability towards undesired hydrolytic processes, both enzymatic and spontaneous, and/or optimal pharmacokinetic properties without needing to chemically modify either the specifier or the drug themselves. In vitro tests thus far carried out on several protease-activatible peptidyl derivative pro-drugs of antineoplastic agents in accordance with the present invention, show a five to seven-fold improvement over the underivatized parent drug in selective cytotoxic activity against malignant cells exhibiting elevated levels of fibrinolytic enzyme activity versus well-matched (displaying similar good sensitivity to the free drug) normal cells not exhibiting such levels of fibrinolytic enzyme activity. These results are indicative of the fact that the peptidyl derivative pro-drugs of the present invention are selective substrates for drug-activating enzymatic cleavage by tumor-associated fibrinolytic enzymes and are selectively activatible to release cytotoxic levels of pharmacologically active drug at sites exhibiting elevated levels of such fibrinolytic enzyme activity. Since normal tissues exhibiting such elevated levels of fibrinolytic enzyme activity are, for the most part, limited to those having a low percentage of replicating cells, peptidyl derivative pro-drugs of antineoplastic agents which are cyto toxic predominantly to rapidly proliferating cells in accordance with the present invention, should be selectively cytotoxic to those malignant cells which exhibit the specific combination of properties of being rapidly proliferating and exhibiting elevated levels of fibrinolytic enzyme activity.

DESCRIPTION OF PREFERRED EMBODIMENTS It will be understood that in the following detailed description and appended claims, the abbreviations and nomenclature employed are those which are standard in amino acid and peptide chemistry, and that all amino acids referred to are in the L-form unless otherwise specified.

The hydrolytic enzyme-activatible pro-drugs in accordance with the present invention may be broadly described as having a molecular structure comprised of a drug moiety and a specifier moiety. The specifier moiety, by means of its chemical structure, targets the pro-drug to one or more species of hydrolytic enzymes and renders the pro-drug a selective substrate for drug-activating enzymatic cleavage by the target hydrolyt enzyme. The drug moiety and the specifier moiety are covalently linked together either directly to form a bipartate molecular structure, or through an intermediate self-immolative connector moiety to form a tripartate molecular structure. In either case, the covalent linkage between the moieties will be such that the drug moiety is rendered pharmacologically inactive, the site of the drug-activating enzymatic cleavage will be at the bond covalently linking the specifier moiety to its immediately adjacent moiety, and the drug-activating enzymatic cleavage will effect release of the drug moiety in pharmacologically active form. The intermediate self-immolative connector moiety, when employed in the pro-drug molecule, has a molecular structure such that the drug-activating enzymatic cleavage of the bond covalently linking it to the specifier moiety will initiate spontaneous cleavage of the bond covalently linking it to the drug moiety, to thereby effect release of the drug moiety in pharmacologically active form.

The peptidyl derivative pro-drugs of antineoplastic agents in accordance with the present invention have an antineoplastic agent as their drug moiety and a peptide as their specifier moiety, and are specifically designed to be selective substrates for drug-activating enzymatic cleavage by one or more tumor-associated proteases selected from the group consisting of fibrinolytic enzymes and blood-coagulating enzymes. Blood-coagulating enzymes are those which are involved in the intrinsic or extrinsic system of fibrin clot formation, and include, but are not necessarily limited to, thrombin, thromboplastin, Factor Va, Factor Vila, Factor Villa, Factor IXa, Factor Xa, Factor XIa, and Factor Xlla. Fibrinolytic enzymes are those which are involved in the physiological mechanism for dissolving fibrin clots, and include plasmin and plasminogen activator. Recent evidence suggests that all of these proteases are associated with a great many tumors, and that plasmin and plasminogen activator, in particular, are present in these tumors at elevated levels sufficient for pro-drug activation. In order to be suitable for conversion into a pro- drug in accordance with the present invention, the antineoplastic agent should be one having an unhindered chemically reactive site whose derivatization will inhibit the pharmacological activity of the antineoplastic agent. Such reactive site will typically be a free amino group or a free hydroxyl group, since these groups are most readily derivatizable with peptides. However, where an intermediate self-immolative connector is employed in forming the pro-drug, the reactive site for derivatization of the antineoplastic agent may also be a free sulfhydryl group. A number of known antineoplastic agents meet the above requirements, including, for example, cytosine arabinoside, adriamycin, daunomycin, 6-thioguanine fluorodeoxyuridine, bis- (2-chloroethyl) amine, phenylene- diamine mustard, 3' -aminothymidine, L-alanosine, 2-amino- thiodiazole, 1 , 4-dihydroxy-5 ,8-bis (2-aminoethylamino) -9 , 10-anthracenedione ,

C The peptide specifier employed for derivatizing the antineoplastic agent so as to convert it into a tumor- specific pro-drug in accordance with the present invention has an amino acid residue sequence specifically tailored so that it will be selectively enzymatically cleaved from the resulting peptidyl derivative pro-drug by one or more of the tumor-associated fibrinolytic and/or blood- coagulating proteases. Examination of the cleavage sites in the natural substrates for these proteases provides a basis for choosing appropriate amino acid residue sequence for the peptide specifier. Since at least most of the fibrinolytic and blood-coagulating proteases appear to hav in common a relatively high degree of specificity toward cleavage sites in their natural substrates which have a basic amino acid residue on the carboxyl side thereof, it is preferred to form the peptide specifier with a basic amino acid residue in its C-terminal position, and to carry out the derivatization of the antineoplastic agent with the C-terminus of the peptide specifier. Suitable basic amino acid residues for use as the C- terminal amino acid residue of the peptide specifier include lysine, arginine, histidine, ornithine, and citrulline, with lysine and arginine being particularly preferred.

The amino acid residue in the position immediately adjacent to the C-terminal amino acid residue also appears to play a significant role in imparting the desired protease-specificity to the peptide specifier. Such penultimate amino acid residue is preferably a hydrophobic amino acid residue or glycine. Suitable hydrophobic amino acid residues include alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan and proline. Alanine, leucine and glycine are particularly preferred amino acid residues for use in such penultimate position of the peptide specifier.

For facilitating preparation of the pro-drug and enhancing its stability against undesired hydrolytic processes, the N-terminal amino acid residue of the peptide specifier is preferably a D-amino acid residue, a protected L-amino acid residue, or protected glycine. Suitable protecting groups are well known in the art of peptide chemistry, and include, for example carbobenzoxy (CBZ) , t-butoxycarbonyl (Boc), p-toluene sulfonyl, and benzoyl, with CBZ being particularly preferred. Most preferably, the N-terminal amino acid residue is a D-amino acid residue, such as, for example, D-valine or D-isoleucine, since this provides the peptide specifier with better solubility properties than with the protected L-amino acid residue or protected glycine. The amino acid residue chain length of the peptide specifier preferably ranges from that of a tripeptide to that of a pentadecapeptide. It will be understood, however, that peptide specifiers as short as dipeptides and longer than pentadecapeptides may also suitably be employed.

With the foregoing basic considerations serving as a general guideline, numerous specific peptide specifier molecules suitable for use in the present invention can be designed and optimized in their selectivity for enzymatic cleavage by a particular one of the tumor-associated fibrinolytic and blood-coagulatin proteases . Based upon the information which is presently available in regard to the cleavage site specificities and the tumor-associated concentrations of these proteases, the presently preferred peptide specifiers for use in the present invention are those which are optimized toward the fibrinolytic proteases, plasmin and plasminogen activator. Its high degree of cleavage site specificity makes plasminogen activator a particularly attractive target protease from the standpoint of designing pro-drugs with optimal selectivity. On the other hand, since one plasminogen activator molecule is capable of converting numerous molecules of plasminogen to plasmin, plasmin will generally have a substantially greater tumor-associated concentration than plasminogen activator and, notwithstanding its lower degree of cleavage site specificity, may be more likely to provide a target large enough to generate pharmacologically significant concentrations of the antineoplastic agent from the pro-drug. In any event, both plasminogen activator, due to its high degree of cleavage site specificity, and plasmin, due to its high tumor-associated concentration, appear to be the target proteases of choic in determining optimal amino acid residue sequences for the peptide specifier under the aforementioned general guidelines.

In the peptide specifiers optimized toward plasmin as the target protease, the C-terminal amino acid residue is preferably lysine, the amino acid residue in the position immediately adjacent to the C-terminal amino acid residue is preferably leucine, and the N-terminal amino acid residue is preferably D-valine or D-isoleucine, Specific examples of this preferred embodiment of peptide specifiers include the tripeptides D-Val-Leu-Lys and

D-Ile-Leu-Lys, and the tetrapeptides D-Val-Ser-Leu-Lys and D-Ile-Ser-Leu-Lys.

In the peptide specifiers optimized toward plasminogen activator as the target protease, the amino acid residue sequence preferably substantially mimics the amino acid residue sequence on the carboxyl side of the Arg-Val bond in plasminogen which serves as the site of cleavage of plasminogen by plasminogen activator, with the C-terminal amino acid residue preferably being arginine, and the amino acid residue in the position immediately adjacent to the C-terminal amino acid residue being glycine. Specific examples of this preferred em¬

Optimization of the peptide specifier toward one or more of the blood-coagulating enzymes as the target tumor-associated protease may similarly be accomplished by choosing an amino acid residue sequence in accordance with the aforementioned general guidelines, but which substantially mimics the amino acid residue sequence on the carboxyl side of the cleavage site in the appropriate natural or known artificial substrates for the particular enzyme. Examples of such substrates are disclosed by Claeson, et al, "Substrate Structure and Activation Relationship", appearing in New Methods For the Analysis of Coagulation Using Chromogenic Substrates, Ed. I. Witt, Walter de Gruyter, Berlin, New York, Pages 37-54 (1977), incorporated herein by reference. Representative peptide specifiers within the scope of the present invention and optimized toward thrombin as the target protease, include the tripeptides p-toluene sulfonyl-Gly-Pro-Arg and benzoyl-Phe-Val-Arg. A representative peptide specifier in accordance with the present invention and optimized toward Factor Xa as the target protease is the tetrapeptide benzoyl-Ile- Glu-Gly-Arg.

In the preferred procedure for synthesizing the peptidyl derivative pro-drugs in accordance with the present invention, the peptide specifier is first separately prepared with its C-terminus in the free acid form, and with all of its other reactive groups suitably blocked. Synthesis of the peptide specifier may be carried out by standard peptide synthesis techniques well known in the art, including either solution-phase or solid-phase methods . Particularly where the peptide being synthesized is of relatively short chain length, the solution-phase methods offer certain advantages in that the peptide is directly prepared in the blocked form needed for the subsequent derivatization of the drug, and the intermediates in the synthesis can be purified, insuring product peptide purity. If solid-phase methods are employed, various known techniques may be used for the removal of the blocked peptide from the resin, for example, by using either photocleavable attachment linkages, or by acyl transfer with 2-dimethylaminoethanol. followed by hydrolysis.

If the antineoplastic agent being converted into a pro-drug contains more than one reactive site on its molecule, those reactive sites other than the one being derivatized may be suitably protected prior to the derivatization reaction. Any of the conventional protecting groups well known in the art may suitably be used for this purpose. For example, in derivatizing the 5'-hydroxyl group of cytosine arabinoside, the 4-amino group of the base may suitably be protected as the Schiffs base using dimethylformamide dimethyl ketal or diisopropylformamide dimethyl ketal, with the latter providing more favorable organic solubility characteristics important in attaining good product recoveries. Deprotection following the derivatization reaction may be effected with trifluoroacetic acid. Another instance where protecting groups might be used to achieve more favorable organic solubility characteristics would be in the derivatization of the 2- amino group of 6-thioguanine, wherein benzylation of the 6-thιo group and at the N4 position will increase the solubility of the parent drug. Deprotection of S and N benzyl protecting groups following the derivatization reaction can be accomplished by treatment with sodium in liquid ammonia.

In preparing the bipartate peptidyl derivative pro-drugs in accordance with the present invention, the peptide specifier, with its C-terminus in the free acid form, and with all of its other reactive groups suitably blocked, is directly reacted with a carboxyl- reactive site of the antineoplastic agent whose derivatization inhibits pharmacological activity, to thereby form a direct covalent linkage between the C- terminus of the peptide specifier and said carboxyl- reactive site of the antineoplastic agent. Such covalent linkage will either be an amide linkage, i.e., when the carboxyl-reactive site is a free amino group; or an ester linkage, i.e., when the carboxyl- reactive site is a free hydroxyl group. Where a choice is available between synthesizing the pro-drug with either an amide linkage or an ester linkage, amide linkages are generally preferred in view of the fact that ester-linked pro-drugs tend to lose at least some selectivity due to hydrolysis by nonspecific esterases. Standard ester-forming and amide-forming techniques well known in the art may be used for carrying out the derivatization reaction. For example, the amide- linked derivatives may suitably be prepared via a mixed anhydride reaction with the aid of isobutyl chloroformate and either triethylamine or N-methyl morpholine in a suitable solvent such as dimethylformamide or dioxane/tetrahydrofuran. Following the derivatization reaction, the protecting groups are removed, for example by treatment with trifluoroacetic acid in methylene chloride, to yield the desired peptidyl derivative pro- drug.

The tripartate pro-drugs in accordance with the present invention employ an intermediate self-immolative connector moiety which spaces and covalently links together the drug moiety and the specifier moiety. Since the self-immolative connector aspect of the present invention is believed to be a novel concept in the design of hydrolytic enzyme-activatible pro-drugs in general, it will be described, first of all, in terms of its broader applications, and thereafter, as it more specifically relates to the peptidyl derivative pro-drugs of antineoplastic agents in accordance with the present invention.

In its broadest sense, a self-immolative connector may be defined as a bifunctional chemical moiety which is capable of (1) covalently linking together two spaced chemical moieties into a normally stable tripartate molecule; (2) releasing one of said spaced chemical moieties from the tripartate molecule by means of an enzymatic cleavage; and (3) following said enzymatic cleavage, spontaneously (i.e., non- enzymatically) cleaving from the remainder of the molecule to release the other of said spaced chemical moieties. As applied to the design of hydrolytic enzyme-activatible pro-drugs in accordance with the present invention, the self-immolative connector is covalently linked at one of its ends to the specifier moiety and covalently linked at its other end to the reactive site of the drug moiety whose derivatization inhibits pharmacological activity, so as to space and covalently link together the specifier moiety and the drug moiety into a tripartate molecule which is stable and pharmacologically inactive in the absence of the target hydrolytic enzyme, but which is enzymatically cleavable by such target hydrolytic enzyme at the bond covalently linking the connector moiety to the specifier moiety to thereby effect release of the specifier moiety from the tripartate molecule. Such enzymatic cleavage, in turn, will activate the self-immolating character of the connector moiety and initiate spontaneous cleavage of the bond covalently linking the connector moiety to the drug moiety, to thereby effect release of the drug in pharmacologically active form. A self-immolative connector offers several potential advantages in hydrolytic enzyme-activatible pro-drug design. First of all, in those instances where the drug molecule being derivatized is large and/or has pronounced polar or apolar character, a bipartate pro-drug formed by a direct linkage of the specifier moiety to the drug moiety may not be a suitable substrate for the target hydrolytic enzyme due to steric and/or electronic hindrance at the intended enzymatic cleavage site caused by the close proximity of such site to the drug molecule. By inserting an intermediate self-immolative connector moiety between the specifier moiety and the drug moiety, the drug-activating enzymatic cleavage site may be spaced sufficiently far away from the drug molecule so as to prevent steric and/or electronic interference with the enzymatic cleavage reaction, and without the connector itself preventing release of the free drug in pharmacologically active form following the enzymatic cleavage reaction. This should allow construction of many more classes and types of hydrolytic enzyme-activatible pro-drugs. Secondly, by varying the functionality of the drug-derivatizing end of the self-immolative connector from that of the specifier, the self-immolative connector may provide greater versatility in the type of linkage used for derivatizing the drug, and may enable linkages which are more stable towards undesired hydrolytic processes (both enzymatic and spontaneous) than are the direct specifier-drug linkages. Thirdly, by providing the self-immolative connector with numerous sites for chemical substitution, it should be possible to design tripartate pro-drugs with optimal pharmacokinetic properties without needing to chejnically modify either the specifier or the drug themselves. This should allow the specifier and drug to be individually optimized for their own particular tasks, e.g., the specifier might be optimized as a substrate for the desired hydrolytic enzyme and made relatively resistant to hydrolysis by undesired hydrolytic enzymes, and the drug might be optimized for specific inhibition of some target enzyme.

A connector moiety which has been found to have all of the above-described characteristics rendering it particularly suitable for use as a self-immolative connector, and the manner in which it is employed in the design of hydrolytic enzyme-activatible tripartate pro-drugs in accordance with the present invention, may be represented by the following general formula:

wherein R1 is hydrogen or one or more substituent groups which are either electron-donating groups or electron-withdrawing groups; R2 and R3 may be the same or different and are each selected from the group consisting of hydrogen, alkyl, phenyl, and phenyl substituted with either electron-donating groups or electron-withdrawing groups; R4 is NH or 0; when R4 is NH, the specifier moiety is selected from the group consisting of a peptide, an amino acid, a carboxylic acid, and phosphoric acid; when R4 is O, the specifier moiety is selected from the group consisting of a carboxylic acid, phosphoric acid, and sulfuric acid; and the drug moiety is a normally pharmacologically active agent having a reactive site whose derivatization inhibits pharmacological activity, said reactive site being selected from the group consisting of a free amino group, a free hydroxyl group and a free sulfhydryl group, the covalent linkage between said drug moiety and its adjacent carbonyl group being at said reactive site so as to inhibit the pharmacological activity of said drug moiety.

In the above definitions of R1, R2 and R3, any of the common electron-donating and electron- withdrawing groups well known in the art may suitably be employed. By way of example suitable electron- donating groups include -NH2, -OH, -OCH3, -NHCOCH3, -C6H5, and -CH3; and suitable electron-withdrawing groups include -NH+(CH3)3, -N02, -CN, -S03H, -COOH, -CHO, -COR, -Cl, -Br, -I, and -F . In the above definitions of R2 and R3 , the alkyl group will generally be a lower alkyl group but may, if desired, be of longer chain length, for example, up to about 15 carbon atoms . Preferred embodiments of the connecto moiety are those in which R1 is hydrogen, R2 is hydrogen or methyl, and R3 is hydrogen or methyl. In Formula I, above, the specifier moiety and the R4 group together constitute a substrate recognitio site for a particular class of target hydrolytic enzymes which is dependent upon the specific combination of specifier moiety and R4 group selected. The various classes of target hydrolytic enzymes for each specific combination of specifier moiety and R. group within the Formula I definitions set forth above, are listed in Table I, below.

The carbonate end of the connector moiety enables derivatization of either a free amino group (by forming a carbamate linkage), a free hydroxyl group (by forming a mixed carbonate linkage), or a free sulfhydryl group (by forming a mono-thio mixed carbonate linkage) on a drug molecule. A wide variety of different types of pharmacologically active agents having one or more of such reactive groups on their molecule will have their pharmacological activity inhibited by derivatization of such reactive groups, and hence would be suitable for use as the drug moiety of Formula I. Representative pharmacologically active agents falling in this category, in addition to the various antineoplastic agents previously listed, are set forth, together with their respective types of pharmacological activity and derivatizable sites for inhibition thereof in Table 2 below.

The theory underlying the mechanism of the self- immolative connector in accordance with the present invention, is explained by the following reaction scheme:

CO2 + Drug

The specifier moiety and R. together act as a group with poor electron-donating capacity. However, enzymatic cleavage of the bond between the specifier moiety and R4 by the target hydrolytic enzyme converts R4 into a strongly electron-donating group, i.e., either NH2 if R4 is NH,or OH if R. is 0. This electron donating effect greatly labilizes the benzylic bond to oxygen, which spontaneously ionizes. Spontaneous decarboxylation of the carbonate anion will then release the drug moiety in pharmacologically active form. The overall release of the drug moiety from the tripartate pro-drug is determined by two processes, namely, (1) the rate of enzymatic hydrolysis of the bond linking the specifier moiety to R4, and (2) the rate of ionization of the bond to the benzylic center. If R1 in Formula I is an electron- donating group, this would tend to decrease process (1) and increase process (2) . On the other hand, if R1 is an electron-withdrawing group, this would tend to increase process (1) and decrease process (2). The net effect of R1 on the rate of release of the drug moiety from the tripartate pro-drug may thus be relatively independent of whether R1 releases or withdraws electrons. The system is thus at least partially buffered against the electronic nature of R1, at least over a certain range. This means thatR1 can be chosen to be either relatively polar, for example, -NH- or -NH+(CH3)3., or relatively non-polar, for example, -C6H5 or -CH3, in order to alter the pharmocokinetic properties of the molecule as desired. The resulting tripartate molecules should not vary greatly in the rate of drug release once they equilibrate with the compartment where the target hydrolytic enzyme acts .R1 may thus be chosen to optimize, for example, log P (1-octanol-water partition coefficient) .

In regard to R„ and R-. in Formula I, the predominant effect of these groups will be on the rate of ionization of the bond to the benzylic center. Electron-donating substituents on these groups will tend to increase the ionization rate, while electron- withdrawing substituents on these groups will tend to decrease the ionization rate. The hydrolytic enzyme-activatible tripartate pro-drugs in accordance with the present invention, may be readily synthesized by, first of all, reacting the specifier with a p-substituted benzyl alcohol reactant having the general formula

*2-,

H-: V >-C-OH (ID

R3

R i |

"^ l wherein R1 , R 2 R3 and R4 have the same meanings as defined above, to obtain a specifier-benzyl alcohol intermediate derivative having the general formula

Specifier-R.-</ \)-C-OH (III)

The specifier-benzyl alcohol intermediate derivative is then reacted with either phosgene or a chloroformate reagent, such as pentafluorophenyl chloro- formate, pentachloropheny.l chloroformate, or p-nitrophenyl chloroformate, to form a second intermediate derivative. This second intermediate derivative will be either a specifier-benzyl chloroformate intermediate derivative, if the reactant is phosgene, or a specifier-benzyl mixed carbonate intermediate derivative, if the reactant is a chloroformate reagent. In either case, such second intermediate derivative is then reacted with the reactive site of the drug whose derivatization inhibits pharmacological activity (i.e., either a free amino group, a free hydroxyl group, or a free sulfhydryl group), to obtain the pro-drug of Formula I. Applying the above-described general procedure to the synthesis of tripartate peptidyl derivative pro-drugs of antineoplastic agents in accordance with the present invention, the peptide specifier, as described in detail hereinabove,with its C-terminus in the free acid form and with all of its other reactive groups suitably blocked with protecting groups, is reacted at its C-terminus with the free amino group of a p-amino benzyl alcohol reactant having the general formula

wherein R- , R„ , R and R . have the same meanings as defined above, to obtain a peptidyl benzyl alcohol having the general formula

This reaction is preferably carried out using a suitable condensing reagent, such as for example, N-ethoxycarbonyl-2-ethoxy-l, 2-dihydroquinoline (EEDQ) in dimethylformamide, to avoid the necessity for protecting the benzylic alcohol function. The peptidyl benzyl alcohol is then reacted with either phosgene or a chloroformate reagent, such as pentafluorophenyl chloroformate, pentachlorophenyl chloroformate, or p-nitrophenyl chloroformate, to convert the peptidyl benzyl alcohol into either a peptidyl benzyl chloroformate (if phosgene is used as the reactant) or a peptidyl benzyl mixed carbonate (if a chloroformate reagent is used as the reactant) . The peptidyl benzyl chloroformate or peptidyl benzyl mixed carbonate is then reacted with a reactive site (a free amino group, a free hydroxyl group, or a free sulfhydryl group) of the antineoplastic agent whose derivatization inhibits pharmacological activity, to obtain a derivatization reaction product from which the protecting groups are then removed, such derivatization reaction product having the general formula

In preparing the peptidyl derivative pro-drugs of antineoplastic agents in accordance with the present invention, the tripartate molecular structure of Formula VI, consisting of the peptide specifier moiety, intermediate self-immolative connective moiety, and trie antineoplastic agent moiety, is particularly advantageous when the antineoplastic agent moiety is adriamycin; daunomyci.n., or bis- (2-chloroethyl) amine, since the molecules of these drugs are such as to tend to cause steric or electronic hindrance problems if the intended drug-activating enzymatic cleavage site is in close proximity to the drug molecule.

These problems are minimized by the spacing provided by the self-immolative connector moiety.

The hydrolytic enzyme-activatible pro-drugs of the present invention, whether of the bipartate or tripartate molecular structure, will generally be administered in the same manner as the parent drug, i.e., orally or parenterally, with parenteral administration, e.g., intravenous, intramuscular or intraarterial, being generally preferred in order to minimize the possibility of premature activation of the pro-drug by non-specific hydrolytic enzymes. The dose levels of the pro-drug should be such as to provide the requisite dose of the free drug. This will generally require that the pro-drug be administered in somewhat larger doses than the parent drug sufficient to allow for the possibility of incomplete activation of the pro-drug into the free drug.

The invention will be further illustrated by way of the following examples.

Example 1 (A) Synthesis of Peptide Specifier

Boc-D-Val was condensed with Leu-OMe ester using dicyclohexylcarbodiimide (DCC) in dimethylformamide/ methylene chloride solution. The resulting di peptide was converted to the free acid by hydrolyzing the ester with NaOH to yield Boc-D-Val-Leu-OH. Nε-CBZ-Nε-Boc-Lys was converted to the methyl ester via treatment with CH2N2 and the amino group freed by hydrogenolysis over Pd/C to yield This latter compound was condensed with the Boc-D-Val-Leu-OH (prepared as above) via a mixed anhydride reaction with isobutylchloroformate in dimethylformamide to yield Boc-D-Val-Leu-Nε-Boc-Lys-OMe. The crude tripeptide was purified by column chromatography on silica gel, and the methyl ester freed by alkaline hydrolysis to yield Boc-D-Val-Leu-Nε-Boc-

Lys-OH.

(B) Synthesis of Bipartate Pro-Drug

The antineoplastic agent AT-125 having the formula

was derivatized at its free amino group by condensing it with the protected peptide specifier Boc-D-Val-Leu-Nε-Boc-Lys-OH (prepared as above) via a mixed anhydride reaction with isobutyl- chloroformate in dioxane/tetrahydrofuran. The resulting product was deprotected with trifluoro acetic acid (TFA) in methylene chloride to yield the desired pro-drug D-Val-Leu-Lys-AT-125. A portion of this peptidyl derivative pro-drug was treated with trypsin in Tris buffer. Two products were obtained which showed Rf on TLC silica plates corresponding to the tri peptide D-Val-Leu-Lys-OH and the free drug AT-125. This demonstrates that the pro-drug is a sub- strate for trypsin and, presumably, other trypsin like proteases such as plasmin and plasminogen activator, which generally show a specificity similar to that of trypsin. A similar result was obtained with plasmin.

Example 2

The peptidyl derivative pro-drug prepared in accordance with Example 1, was tested for its tumor- specific cytotoxic activity by means of an j-n vitro test procedure utilizing a cell culture system with well-matched normal and malignant cells which differed substantially in fibrinolytic enzyme (i-e_., plasmin and plasminogen activator) levels but displayed similar good sensitivity to the free drug. The cell culture system employed was chick embryo fibroblasts, both normal and transformed with Rous Sarcoma virus. The transformed cells exhibit a substantially higher level of fibrinolytic enzyme activity than the normal cells. The test procedure was carried out as follows. Chick embryo fibroblasts, either normal (N) or transformed with Rous Sarcoma virus (SR) were plated in 35 mm plastic dishes at an initial titer of 1.5 x

10 5 cells per dish (N) or 3 x 105 cells per dish (SR) . Allowing for the difference in plating efficiencies between normal and transformed cells, this leads to similar numbers of cells per plate.

The medium used was Dulbecco's Modified Minimal

Eagle's Medium (DME) supplemented with 10% Tryptose- Phosphate broth, 4% calf serum, and 1% chick serum. Cells grow exponentially in this medium with a doubling time of 18 to 20 hours. After 24 hours the cells were changed to DME medium containing 5% chick serum and lacking Tryptose-Phosphate broth and calf serum. After a further 18 hours, the test drugs (which included both the peptidyl derivative pro-drug and, for purposes of comparison, the underivatized parent drug) were added at various concentrations for a further 5 hours. Finally, in order to measure DNA synthesis 3H-thymidine at a final concentration of 1 microcurie/ml was added to each dish for 30 minutes. The medium was removed and the cells were incubated with cold 5% trichloroacetic acid for a further 15 minutes. After 3 further washes with cold trichloroacetic acid, the DNA was hydrolyzed with 10% trichloroacetic acid at 70 °C for two hours. The solubilized material was counted in an Omnifluor- Triton-X-100 scintillation fluid. Control experiments have demonstrated that this measurement of thymidine incorporation into DNA corresponds to cell numbers as determined in a Coulter counter. By means of plotting the residual 3H-thymidine incorporation as a function of drug concentration compared to untreated control, the ED50 (i.e. , the drug concentration at which incorporation of thymidine is reduced to 50% of the untreated control) is determined for each of the two test drugs (i.e., the peptidyl derivative pro-drug and the corresponding underivatized parent drug) against both the normal cells and the transformed cells. The therapeutic index of each drug (i.e., the ratio of its pharmacological activity to its toxicity) is then determined as the ratio of its EDo,.u-, against the normal cells to its ED50 against the transformed cells. Following this procedure, the therapeutic index for the peptidyl derivative pro-drug in accordance with the present invention was determined to be 5.3, in comparison to a therapeutic index of 1.2 for the corresponding underivatized parent drug. Thus, the peptidyl derivative pro-drug in accordance with the present invention exhibits an approximately 5-fold improvement in therapeutic index over the corresponding underivatized parent drug.

The above-described test results are indicative of the fact that the peptidyl derivative pro-drugs of the present invention are selective substrates for drug-activating enzymatic cleavage by tumor- associated fibrinolytic enzymes, and are selectively activatible to release cytotoxic levels of pharmacologically active drug at sites exhibiting elevated levels of such fibrinolytic enzyme activity.

Example 3 Employing a synthesis procedure similar to that described in Example 1, above, the antineoplastic agent, phenylenediamine mustard, was derivatized at its free amino group with D-Val-Leu-Lys peptide specifier to convert it into the peptidyl derivative pro-drug D-Val-Leu-Lys-phenylenediamine mustard. When tested by means of the in vitro assay described in Example 2, above, the D-Val-Leu-Lys-phenylenediamine mustard pro-drug showed a 7-fold improvement in its therapeutic index in comparison with the underivatized phenylenediamine mustard parent drug.

Example 4 Employing a synthesis procedure similar to that described in Example 1, above, the antineoplastic agent, adriamycin, was derivatized at its free amino group on daunosamine with the D-Val-Leu-Lys peptide specifier to convert it into D-Val-Leu-Lys-adriamycin pro-drug. When tested by means of the in vitro assay described in Example 2, above, the D-Val-Leu-Lys-adriamycin pro-drug showed a 6-fold improvement in its therapeutic index, in comparison with the underivatized adriamycin parent drug.

Example 5 The feasibility of the self-immolative connector aspect of the present invention in the design of hydrolytic enzyme-activatible tripartate pro-drugs was demonstrated by means of the following model study which utilized p-nitroaniline as the "drug" because of the ease of colorimetric detection.

Synthesis of the tripartate pro-drug model was carried out as follows. N -t-Boc-N -TFA lysine was coupled to p-aminobenzyl alcohol using N-ethoxycarbonyl- 2-ethoxy-l, 2-dihydroquinoline (EEDQ) in dimethylformamide. The product was purified by crystallization from ethyl acetate: ether. Next this material was reacted with p-nitrophenylisocyanate in dry pyridine to yield

which was purified by preparative TLC on silica. Finally, the TFA group was removed with tetramethyl- guanidine in acetonitrile-water 1:1 to yield the tripartate pro-drug model having the formula

Th final structure was characterized by NMR and IR spectroscopy.

In order to demonstrate the possibility of self- immolation with this tripartate pro-drug model, it was dissolved in 1 ml 50 mM Bis-Tris-Cl buffer pH 6.7 at 1 mM and treated with 2 μg of trypsin. There was an instantaneous release of p-nitroaniline as measured by the increase of optical density at 405 nm. The rate of release of p-nitroaniline was comparable to the rate of release expected for cleavage of the

Lys-anilide bond, and no lag was noted. TLC analysis showed the expected products Nα-Boc-Lys-OH, p-aminobenzy alcohol, and p-nitroaniline. The release of p-nitroaniline was blocked by prior treatment of trypsin with tosyl-lysyl chloromethyl ketone (TLCK) .

Two similar tripartate pro-drug models were similarly synthesized, one with one of the benzylic hydrogens replaced by a methyl group, and the other with p-nitroaniline replaced by aniline. In both cases, TLC analysis again showed that trypsin hydrolysis released the expected products, including p-nitroaniline or aniline. This was confirmed spectrophotometrically in the case of the p-nitroaniline derivative.

The above-described model studies demonstrate the feasibility under physiological conditions of the self-immolation of the intermediate connector moiety in the hydrolytic enzyme-activatible tripartate pro-drugs in accordance with the present invention. While the fibrinolytic and blood-coagulating enzyme- activatible peptidyl derivative pro-drugs in accordance with the present invention have been described with particular reference to the preferred embodiments thereof wherein the drug moiety is an antineoplastic agent, it will be understood that the drug moiety, in either the bipartate or tripartate structure, could be any normally pharmacologically active compound which is suitably convertible into a pro-drug by derivatization with the peptide specifiers described above and whose site of intended action is known to exhibit elevated levels of fibrinolytic and/or blood-coagulating enzyme activity. By way of example, elevated levels of fibrinolytic and/ or blood-coagulating enzymes are normally exhibited in the skin, which is the site of intended action of antipsoriasis agents, such as fluocinonide; in the joint, which is the site of intended action of anti-arthritic agents, such as betamethasone; and in the uterus, which is the site of intended action of antifertility or implantation agents such as estrogenic and progestational steroids. Any of these drugs could be suitably derivatized with the peptide specifiers as described above to convert them into peptidyl pro-drugs, of either the bipartate or tripartate structure, which are selective substrates for fibrinolytic and/or blood-coagulating proteases so as to be selectively activatible at their site of intended action.

Claims

1. A tumor-specific pro-drug of an antineoplastic agent comprising a peptidyl derivative of said antineoplastic agent, said peptidyl deriva tive being a selective substrate for drug-activating enzymatic cleavage by one or more tumor-associated proteases selected from the group consisting of fibrinolytic enzymes and blood-coagulating enzymes.
2. The pro-drug of Claim 1, wherein said tumor-associated protease is a fibrinolytic enzyme selected from the group consisting of plasmin and plasminogen activator.
3. The pro-drug of Claim 1, wherein said tumor-associated protease is a blood-coagulating enzyme selected from the group consisting of thrombin, thrombo plastin. Factor Va, Factor Vila, Factor Villa, Factor IXa,Factor Xa, Factor XIa, and Factor Xlla.
4. The pro-drug of Claim 1, wherein the molecular structure of said peptidyl derivative is comprised of a peptide specifier moiety and an antineoplastic agent moiety, said two moieties being covalently linked together either directly or through an intermediate self-immolative connector moiety in a manner such that said antineoplastic agent moiety is rendered pharmacologically inactive, the site of said drug-activating enzymatic cleavage will be at the bond covalently linking said peptide specifier moiety to its immediately adjacent moiety, and said drug-activating enzymatic cleavage will effect release of said antineoplastic agent moiety in pharmacologically active form.
5. The pro-drug of Claim 4, wherein the covalent linkage of said peptide specifier moiety to its immediately adjacent moiety is at the C- terminus of said peptide specifier moiety, and the C-terminal amino acid residue of said peptide specifier moiety is a basic amino acid residue.
6. The pro-drug of Claim 5, wherein said C-terminal amino acid residue is arginine or lysine.
7. The pro-drug of Claim 5, wherein said peptide specifier moiety contains a hydrophobic amino acid residue or glycine in the position immediately adjacent to said C-terminal amino acid residue.
8. The pro-drug of Claim 7 , wherein said hydrophobic amino acid residue is selected from the group consisting of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan and proline.
9. The pro-drug of Claim 5 , wherein the N-terminal amino acid residue of said peptide specifier moiety is a D-amino acid residue, a protected L-amino acid residue, or protected glycine.
10. The pro-drug of Claim 5, wherein said peptide specifier moiety has an amino acid residue chain length ranging from that of a tripeptide to that of a pentadecapeptide.
11. The pro-drug of Claim 10, wherein said peptide specifier moiety contains a hydro- phobic amino acid residue or glycine in the position immediately adjacent to said C-terminal amino acid residue, and the N-terminal amino acid residue of said peptide specifier moiety is a D-amino acid, residue, a protected L-amino acid residue or protected glycine.
12. The pro-drug of Claim 10, wherein the amino acid residue sequence in said peptide specifier moiety is such that its C-terminal amino acid residue is arginine or lysine, its amino acid residue in the position immediately adjacent to said C-terminal amino acid residue is alanine, leucine, or glycine, and its N-terminal amino acid residue is a D-amino acid residue, a protected L-amino acid residue or protected glycine.
13. The pro-drug of Claim 12, wherein said
C-terminal amino acid residue is lysine, said amino acid residue in the position immediately adjacent to said C-terminal amino acid residue is leucine, and said N-terminal amino acid residue is D-valine or D-isoleucine.
14. The pro-drug of Claim 12, wherein the amino acid residue sequence in said peptide specifier moiety substantially mimics the amino acid residue sequence on the carboxyl side of the Arg-Val bond in plasminogen which serves as the site of cleavage of plasminogen by plasminogen activator, with said C-terminal amino acid residue being arginine, and said amino acid residue in the position immediately adjacent to said C-terminal amino acid residue being glycine.
15. The pro-drug of Claim 4, wherein said peptidyl derivative has a bipartate molecular structure consisting of said peptide specifier moiety and said antineoplastic agent moiety, said two moieties being directly covalently linked together by means of a covalent linkage formed between the C-terminus of said peptide specifier moiety and a carboxyl-reactive site of said antineoplastic agent moiety whose derivatization inhibits pharmacological activity.
16. The pro-drug of Claim 15, wherein said covalent linkage is an amide linkage formed between the C-terminus of said peptide specifier moiety and a free amino group of said antineo- plastic agent moiety.
17. The pro-drug of Claim 15, wherein said covalent linkage is an ester linkage formed between the C-terminus of said peptide specifier moiety and a free hydroxyl group of said antineoplastic agent moiety.
18. The pro-drug of Claim 4, wherein said peptidyl derivative has a tripartate molecular structure consisting of said peptide specifier moiety, said intermediate self-immolative connector moiety and said antineoplastic agent moiety, said intermediate self-immolative connector moiety being covalently linked at its one end to the C- terminus of said peptide specifier moiety and covalently linked at its other end to a reactive site of said antineoplastic agent moiety whose derivatization inhibits pharmacological activity, said intermediate self-immolative connector moiety having a molecular structure such that said drug- activating enzymatic cleavage of the bond covalently linking it to said peptide specifier moiety will initiate spontaneous cleavage of the bond covalently linking it to said antineoplastic agent moiety to thereby effect release of said antineoplastic agent moiety in pharmacologically active form.
19. The pro-drug of Claim 18 , wherein said intermediate self-immolative connector moiety has the general formula :
wherein R. is hydrogen or one or more substituent groups which are either electron-donating groups or electron-withdrawing groups, and R2 and. R3 may be the same or different and are each selected from the group consisting of hydrogen, alkyl, phenyl, and phenyl substituted with either electron-donating groups or electron-withdrawing groups, said connector moiety having its terminal amino group covalently linked to the C- terminus of said peptide specifier moiety and its terminal carbonyl group covalently linked to said reactive site of said antineoplastic agent moiety.
20. The pro-drug of Claim 19, whereinR1 is hydrogen, R2 is hydrogen or methyl, and Ri3is hydrogen or methyl.
21. The pro-drug of Claim 19, wherein said reactive site of said antineoplastic agent moiety is a free amino group, a free hydroxyl group, or a free sulfhydryl group.
22. The pro-drug of Claim 21, wherein said antineoplastic agent moiety is selected from the group consisting of adriamycin, daunomycin, and bis- (2- chloroethy1) amine.
23. The pro-drug of Claim 4, wherein the covalent linkage of said antineoplastic agent moiety to its immediately adjacent moiety is at a reactive site of said antineoplastic agent moiety whose derivatization inhibits pharmacological activity.
24. The pro-drug of Claim 23, wherein said reactive site of said antineoplastic agent moiety is a free amino group, a free hydroxyl group, or a free sulfhydryl group.
25. The pro-drug of Claim 24, wherein said antineoplastic agent moiety is selected from the group consisting of cytosine arabinoside, adriamycin, daunomycin, 6-thioguanine, fluorodeoxyuridine, bis- (2- chloroethyl) amine, phenylenediamine mustard, 3'-amino- thymidine, L-alanosine, 2-aminothiodiazole, 1,4-dihydroxy-
5 ,8-bis (2-aminoethylamino) -9,10-anthracenedione,
26. The pro-drug of Claim 25, wherein said peptide specifier moiety has an amino acid residue chain length ranging from that of a tripeptide to that of a pentadecapeptide, the covalent linkage of said peptide specifier moiety to its immediately adjacent moiety is at the C-terminus of said peptide specifier moiety, and the amino acid residue sequence in said peptide specifier moiety is such that its C- terminal amino acid residue is a basic amino acid residue, its amino acid residue in the position immediately adjacent to said C-terminal amino acid residue is a hydrophobic amino acid residue or glycine, and its N-terminal amino acid residue is a D-amino acid residue, a protected L-amino acid residue or protected glycine.
27. A method of rendering an antineoplastic agent tumor-specific which comprises derivatizing said antineoplastic agent either directly or through an intermediate self-immolative connector with a peptide specifier at a reactive site appropriate for inhibiting the pharmacological activity of said antineoplastic agent, said peptide specifier having an amino acid residue sequence such that it will be selectively enzymatically cleaved from said antineoplastic agent by one or more tumor-associated proteases selected from the group consisting of fibrinolytic enzymes and blood- coagulating enzymes so as to effect release of said antineoplastic agent in pharmacologically active form.
28. The method of Cliam 27, wherein said tumor-associated protease is a fibrinolytic enzyme selected from the group consisting of plasmin and plasminogen activator.
29. The method of Claim 27, wherein said reactive site is a free amino group, a free hydroxyl group, or a free sulfhydryl group.
30. The method of Claim 29, wherein said antineoplastic agent is selected from the group consisting of cytosine arabinoside, adriamycin, daunomycin, 6-thioguanine, fluorodeoxyuridine, bis- (2-chloroethyl) amine, phenylenediamine mustard, 3'-aminothymidine,
L-alanosine, 2-aminothiodiazole, l,4-dihydroxy-5,8-bis (2-aminoethylamino) -9, 10-anthracenedione,
31. The method of Claim 27, wherein said derivatization is carried out either directly or through an intermediate self-immolative connector with the C-terminus of said peptide specifier, and the C-terminal amino acid residue of said peptide specifier is a basic amino acid residue.
32. The method of Claim 31, wherein said peptide specifier has an amino acid residue chain length ranging from that of a tripeptide to that of a pentadecapeptide, and its N-terminal amino acid residue is a D-amino acid residue, a protected L-amino acid residue or protected glycine.
33. The method of Claim 32, wherein said peptide specifier contains a hydrophobic amino acid residue or glycine in the position immediately adjacent to said C-terminal amino acid residue.
34. The method of Claim 33, wherein said C-terminal amino acid residue is lysine, said amino acid residue in the position immediately adjacent to said C-terminal amino acid residue is leucine, and said N-terminal amino acid residue is D-valine or D-isoleucine.
35. The method of Claim 33, wherein the amino acid residue sequence in said peptide speci fier substantially mimics the amino acid residue sequence on the carboxyl side of the Arg-Val bond in plasminogen which serves as the site of cleavage of plasminogen by plasminogen activator, with said C-terminal amino acid residue being arginine, and the amino acid residue in the position immediately adjacent to said C-terminal amino acid residue being glycine.
36. The method of Claim 27, wherein the peptide specifier reactant in the derivatization reaction has its C-terminus in the free acid form and all of its other reactive groups suitably blocked with protecting groups, said reactive site is a free amino group or a free hydroxyl group of said antineoplastic agent, said derivatization reaction is carried out directly between said C-terminus of said peptide specifier reactant and said reactive site of said antineoplastic agent, and subsequent to said derivatization reaction said protecting groups are removed from the reaction product.
37. The method of Claim 27, wherein the peptide specifier, reactant in the derivatization reaction has its C-terminus in the free acid form and all of its other reactive groups suitably blocked with protecting groups; said reactive site is a free amino group, a free hydroxyl group or a free sulfhydryl group of said antineoplastic agent; said derivatization reaction is carried out in a multi-step procedure comprising:
(a) reacting said C-terminus of said peptide specifier reactant with the free amino group of a p-aminobenzyl alcohol reactant having the general formula
wherein R1 is hydrogen or one or more substituent groups which are either electron-donating groups or electron-withdrawing groups, and R2 and R3 may be the same or different and are each selected from the group consisting of hydrogen, alkyl, phenyl, and phenyl substituted with either electron-donating groups or electron- withdrawing groups, to obtain a peptidyl benzyl alcohol having the general formula
(b) reacting said peptidyl benzyl alcohol with either phosgene or a chloroformate reagent to convert said peptidyl benzyl alcohol into, respectively, either a peptidyl benzyl chloroformate or a peptidyl benzyl mixed carbonate, and
(c) reacting said peptidyl benzyl chloroformate or peptidyl benzyl mixed carbonate with said reactive site of said antineoplastic agent to obtain a derivatization re action product having the general formula
and subsequen o sa er va za on react on sa protecting groups are removed from said derivatization reaction product.
38. The method of Claim 37, wherein R1 is hydrogen, R2 is hydrogen or methyl, and R3 is hydrogen or methyl.
39. The method of Claim 37, wherein said chloroformate reagent is selected from the group consisting of pentafluorophenyl chloroformate, pentachlorophenyl chloroformate, and p-nitrophenyl chloroformate .
40. In a method for the treatment of neoplastic diseases in an animal or human host which comprises administering to said host in need thereof an antineoplastically effective amount of an antineoplastic agent, the improvement consisting of administering said antineoplastic agent in the form of the peptidyl derivative pro-drug of Claim 1, whereby the selectivity of delivery of said antineoplastic agent to the tumor site of intended action is enhanced.
41. A hydrolytic enzyme-activatible pro-drug having the general formula
wherei R1 is hydrogen or one or more substituent groups which are either electron-donating groups or electron- withdrawing groups; R2 and R3 may be the same or different and are each selected from the group consisting of hydrogen, alkyl, phenyl, and phenyl substituted with either electron-donating groups or electron-withdrawing groups; R4 is NH or O; when R4 is NH, the specifier moiety is selected from the group consisting of a peptide, an amino acid, a carboxylic acid, and phosphoric acid; when R4 is 0, the specifier moiety is selected from the group consisting of a carboxylic acid, phosphoric acid, and sulfuric acid; and the drug moiety is a normally pharmacologically active agent having a reactive site whose derivatization inhibits pharmacological activity, said reactive site being selected from the group consisting of a free amino group, a free hydroxyl group and a free sulfhydryl group, the covalent linkage between said drug moiety and its adjacent carbonyl group being at said reactive site so as to inhibit the pharmacological activity of said drug moiety.
42. The pro-drug of Claim 41, wherein R1 is hydrogen, R2 is hydrogen or methyl, and R3 is hydrogen or methyl.
43. The pro-drug of Claim 41, wherein said drug moiety is an antineoplastic agent.
44. The pro-drug of Claim 41, wherein R. is NH and the specifier moiety is a peptide.
45. The pro-drug of Claim 42, wherein said drug moiety is an antineoplastic agent,R4 is NH and the specifier moiety is a peptide.
46. In a method of converting a pharmacologically active drug into a hydrolytic enzyme- activatible pro-drug by derivatizing said drug at a reactive site thereof appropriate for inhibiting its pharmacological activity, with a specifier designed to be selectively enzymatically cleaved from the resulting pro-drug by a target hydrolytic enzyme so as to effect release of said drug in pharmacologically active form, the improvement consisting of spacing the specifier moiety from the drug moiety by means of an intermediate self- immolative connector moiety covalently linked at its one end to said specifier moiety and covalently linked at its other end to said reactive site of said drug moiety, said intermediate self-immolative connector moiety having a molecular structure such that the drug- activating enzymatic cleavage of the bond covalently linking it to said specifier moiety will initiate spontaneous cleavage of the bond covalently linking it to said drug moiety to thereby effect release of said drug in pharmacologically active form.
47. The method of Claim 46, wherein said reactive site of said drug is selected from the group consisting of a free amino group, a free hydroxyl group, and a free sulfhydryl group,and said specifier is selected from the group consisting of a peptide, an amino acid, a carboxylic acid, phosphoric acid and sulfuric acid.
48. The method of Claim 47, wherein said resulting pro-drug has the general formula
wherein R, is hydrogen or one or more substituent groups which are either electron-donating groups or electron-withdrawing groups; R2 and R3 may be the same or different and are each selected from the group consisting of hydrogen, alkyl, phenyl, and phenyl substituted with either electron-donating groups or electron-withdrawing groups; R4 is NH or 0; when R4 is NH, the specifier moiety is selected from the group consisting of a peptide, an amino acid, a carboxylic acid, and phosphoric acid; and when R4 is O, the specifier moiety is selected from the group consisting of a carboxylic acid, phosphoric acid, and sulfuric acid.
49. The method of Claim 48, wherein the preparation of said pro-drug comprises the.steps of: (a) reacting said specifier with a p- substituted benzyl alcohol reactant having the general formula
wherein R1, R2,R3 and R4 have the same meanings as defined above, to obtain a specifier-benzyl alcohol intermediate derivative having the general formula
(b) reacting said specifier-benzyl alcohol intermediate derivative with either phosgene or a chloroformate reagent to convert said specifier-benzyl alcohol intermediate derivative into, respectively, either a specifier-benzyl chloroformate intermediate derivative or a specifier-benzyl mixed carbonate intermediate derivative; and
(c) reacting said specifier-benzyl chloroformate intermediate derivative or specifier-benzyl mixed carbonate intermediate derivative with said reactive site of said drug to obtain said pro-drug.
50. The method of Claim 49, wherein said chloroformate reagent,.is selected from the group consisting of pentTafluorophenyl chloroformate, penta- chlor^phenyl chϊdroformate, and p-nitrophenyl chloroformate.
51. The method of Claim 49, wherein R1 is hydrogen, R2 is hydrogen or methyl, and R3 is hydrogen or methyl.
52. The method of Claim 49, wherein said drug is an antineoplastic agent.
53. The method of Claim 49, wherein R4 is NH and the specifier is a peptide.
54. The method of Claim 51, wherein said drug is an antineoplastic agent, R4 is NH, and the specifier is a peptide.
55. A pro-drug of a normally pharmacologically active compound comprising a peptidyl derivative of said compound, said peptidyl derivative being a selective substrate for drug-activating enzymatic cleavage for one or more proteases selected from the group consisting of fibrinolytic enzymes and blood- coagulating enzymes.
56. The pro-drug of Claim 55, wherein said protease is a fibrinolytic enzyme selected from the group consisting of plasmin and plasminogen activator
PCT/US1980/001290 1979-10-18 1980-10-01 Hydrolytic enzyme-activatible pro-drugs WO1981001145A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US8609679 true 1979-10-18 1979-10-18
US86096 2002-02-27

Publications (1)

Publication Number Publication Date
WO1981001145A1 true true WO1981001145A1 (en) 1981-04-30

Family

ID=22196237

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1980/001290 WO1981001145A1 (en) 1979-10-18 1980-10-01 Hydrolytic enzyme-activatible pro-drugs

Country Status (3)

Country Link
EP (1) EP0038357A4 (en)
CA (1) CA1158557A (en)
WO (1) WO1981001145A1 (en)

Cited By (183)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0237082A3 (en) * 1986-03-14 1988-09-14 Syntex (U.S.A.) Inc. Transglutaminase inhibitors
US4912120A (en) * 1986-03-14 1990-03-27 Syntex (U.S.A.) Inc. 3,5-substituted 4,5-dihydroisoxazoles as transglutaminase inhibitors
US4921941A (en) * 1987-07-01 1990-05-01 Schering Corporation Orally active antiandrogens
US4929630A (en) * 1986-03-14 1990-05-29 Syntex (U.S.A.) Inc. Transglutaminase inhibitors
DE4236237A1 (en) * 1992-10-27 1994-04-28 Behringwerke Ag Prodrugs, their preparation and use as medicaments
EP0642799A1 (en) * 1993-09-09 1995-03-15 BEHRINGWERKE Aktiengesellschaft Improved prodrugs for enzyme mediated activation
EP0648503A1 (en) * 1993-09-22 1995-04-19 BEHRINGWERKE Aktiengesellschaft Pro-prodrugs, their production and use
WO1996001653A1 (en) * 1994-07-11 1996-01-25 Board Of Regents, The University Of Texas System Methods and compositions for the specific coagulation of vasculature
US5561119A (en) * 1991-04-30 1996-10-01 Laboratoires Hoechst Glycosylated prodrugs, their method of preparation and their uses
US5660827A (en) * 1992-03-05 1997-08-26 Board Of Regents, The University Of Texas System Antibodies that bind to endoglin
US5855866A (en) * 1992-03-05 1999-01-05 Board Of Regenis, The University Of Texas System Methods for treating the vasculature of solid tumors
US5863538A (en) * 1992-03-05 1999-01-26 Board Of Regents, The University Of Texas System Compositions for targeting the vasculature of solid tumors
US5877289A (en) * 1992-03-05 1999-03-02 The Scripps Research Institute Tissue factor compositions and ligands for the specific coagulation of vasculature
US5880131A (en) * 1993-10-20 1999-03-09 Enzon, Inc. High molecular weight polymer-based prodrugs
US5965566A (en) * 1993-10-20 1999-10-12 Enzon, Inc. High molecular weight polymer-based prodrugs
US6004555A (en) * 1992-03-05 1999-12-21 Board Of Regents, The University Of Texas System Methods for the specific coagulation of vasculature
US6036955A (en) * 1992-03-05 2000-03-14 The Scripps Research Institute Kits and methods for the specific coagulation of vasculature
US6093399A (en) * 1992-03-05 2000-07-25 Board Of Regents, The University Of Texas System Methods and compositions for the specific coagulation of vasculature
US6153655A (en) * 1998-04-17 2000-11-28 Enzon, Inc. Terminally-branched polymeric linkers and polymeric conjugates containing the same
US6214345B1 (en) 1993-05-14 2001-04-10 Bristol-Myers Squibb Co. Lysosomal enzyme-cleavable antitumor drug conjugates
WO2002022833A1 (en) * 2000-09-15 2002-03-21 Universität Stuttgart Fusion protein from antibody cytokine-cytokine inhibitor (selectokine) for use as target-specific prodrug
WO2002030463A2 (en) 2000-10-12 2002-04-18 Genentech, Inc. Reduced-viscosity concentrated protein formulations
EP1243276A1 (en) * 2001-03-23 2002-09-25 Patrick Henry Beusker Elongated and multiple spacers containing activatible prodrugs
US6511649B1 (en) 1998-12-18 2003-01-28 Thomas D. Harris Vitronectin receptor antagonist pharmaceuticals
US6511648B2 (en) 1998-12-18 2003-01-28 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6524553B2 (en) 1998-03-31 2003-02-25 Bristol-Myers Squibb Pharma Company Quinolone vitronectin receptor antagonist pharmaceuticals
US6537520B1 (en) 1998-03-31 2003-03-25 Bristol-Myers Squibb Pharma Company Pharmaceuticals for the imaging of angiogenic disorders
US6548663B1 (en) 1998-03-31 2003-04-15 Bristol-Myers Squibb Pharma Company Benzodiazepine vitronectin receptor antagonist pharmaceuticals
US6558649B1 (en) 1998-12-18 2003-05-06 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6569402B1 (en) 1998-12-18 2003-05-27 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6749853B1 (en) 1992-03-05 2004-06-15 Board Of Regents, The University Of Texas System Combined methods and compositions for coagulation and tumor treatment
US6794518B1 (en) 1998-12-18 2004-09-21 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
EP1619250A1 (en) 1996-01-08 2006-01-25 Genentech, Inc. WSX receptor and ligands
WO2006009901A2 (en) 2004-06-18 2006-01-26 Ambrx, Inc. Novel antigen-binding polypeptides and their uses
EP1650220A2 (en) 1997-04-07 2006-04-26 Genentech, Inc. Anti-VEGF antibodies
US7037898B2 (en) 1994-08-19 2006-05-02 Laregion Wallone Tumor-activated prodrug compounds and treatment
WO2006074418A2 (en) 2005-01-07 2006-07-13 Diadexus, Inc. Ovr110 antibody compositions and methods of use
WO2006089133A2 (en) 2005-02-15 2006-08-24 Duke University Anti-cd19 antibodies and uses in oncology
US7115573B2 (en) 2000-06-14 2006-10-03 Medarex, Inc. Prodrug compounds with an isoleucine
US7173115B2 (en) 2000-01-13 2007-02-06 Genentech, Inc. Stra6 polypeptides
WO2008011081A2 (en) 2006-07-19 2008-01-24 The Trustees Of The University Of Pennsylvania Wsx-1/p28 as a target for anti-inflammatory responses
EP1950300A2 (en) 1998-11-18 2008-07-30 Genentech, Inc. Antibody variants with higher binding affinity compared to parent antibodies
US7446196B2 (en) 2004-06-03 2008-11-04 Kosan Biosciences, Incorporated Leptomycin compounds
EP1992643A2 (en) 2001-06-20 2008-11-19 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2011886A2 (en) 2002-04-16 2009-01-07 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2014303A2 (en) 2000-07-27 2009-01-14 Genentech, Inc. APO-2L receptor agonist and CPT-11 synergism
WO2009028158A1 (en) 2007-08-24 2009-03-05 Oncotherapy Science, Inc. Dkk1 oncogene as therapeutic target for cancer and a diagnosing marker
EP2042517A1 (en) 2002-09-27 2009-04-01 Xencor, Inc. Optimized FC variants and methods for their generation
EP2052742A1 (en) 2000-06-20 2009-04-29 Biogen Idec Inc. Treatment of B-cell associated diseases such as malignancies and autoimmune diseases using a cold anti-CD20 antibody/radiolabeled anti-CD22 antibody combination
EP2058334A2 (en) 1998-06-12 2009-05-13 Genentech, Inc. Monoclonal antibodies, cross-reactive antibodies and method for producing the same
EP2062916A2 (en) 2003-04-09 2009-05-27 Genentech, Inc. Therapy of autoimmune disease in a patient with an inadequate response to a TNF-Alpha inhibitor
EP2062591A1 (en) 2005-04-07 2009-05-27 Novartis Vaccines and Diagnostics, Inc. CACNA1E in cancer diagnosis detection and treatment
US7541330B2 (en) 2004-06-15 2009-06-02 Kosan Biosciences Incorporated Conjugates with reduced adverse systemic effects
EP2065467A2 (en) 2001-02-22 2009-06-03 Genentech, Inc. Anti-interferon-alpha antibodies
EP2067472A1 (en) 2002-01-02 2009-06-10 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2083088A2 (en) 2005-04-07 2009-07-29 Novartis Vaccines and Diagnostics, Inc. Cancer-related genes
EP2110138A1 (en) 1999-08-27 2009-10-21 Genentech, Inc. Dosages for treatment of anti-erbB2 antibodies
EP2112167A2 (en) 1999-06-25 2009-10-28 Genentech, Inc. Humanized ANTI-ERBB2 antibodies and treatment with ANTI-ERBB2 antibodies
WO2010001585A1 (en) 2008-06-30 2010-01-07 Oncotherapy Science, Inc. Anti-cdh3 antibodies labeled with radioisotope label and uses thereof
EP2143438A1 (en) 2001-09-18 2010-01-13 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2161283A1 (en) 2003-11-17 2010-03-10 Genentech, Inc. Compositions comprising antibodies against CD79b conjugated to a growth inhibitory agent or cytotoxic agent and methods for the treatment of tumor of hematopoietic origin
WO2010056804A1 (en) 2008-11-12 2010-05-20 Medimmune, Llc Antibody formulation
WO2010060920A1 (en) 2008-11-27 2010-06-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Cxcl4l1 as a biomarker of pancreatic cancer
WO2010077634A1 (en) 2008-12-09 2010-07-08 Genentech, Inc. Anti-pd-l1 antibodies and their use to enhance t-cell function
EP2221316A1 (en) 2005-05-05 2010-08-25 Duke University Anti-CD19 antibody therapy for autoimmune disease
WO2010102175A1 (en) 2009-03-05 2010-09-10 Medarex, Inc. Fully human antibodies specific to cadm1
WO2010120561A1 (en) 2009-04-01 2010-10-21 Genentech, Inc. Anti-fcrh5 antibodies and immunoconjugates and methods of use
EP2248829A1 (en) 2003-05-30 2010-11-10 Genentech, Inc. Treatment with anti-VEGF antibodies
EP2263691A1 (en) 2002-07-15 2010-12-22 Genentech, Inc. Treatment of cancer with the recombinant humanized monoclonal anti-erbb2 antibody 2C4 (rhuMAb 2C4)
WO2011017249A1 (en) 2009-08-03 2011-02-10 Medarex, Inc. Antiproliferative compounds, conjugates thereof, methods therefor, and uses thereof
WO2011031397A1 (en) 2009-08-06 2011-03-17 Genentech, Inc. Method to improve virus removal in protein purification
EP2301580A1 (en) 1997-04-07 2011-03-30 Genentech, Inc. Anti-VEGF antibodies
WO2011038302A2 (en) 2009-09-25 2011-03-31 Xoma Technology Ltd. Novel modulators
WO2011038301A2 (en) 2009-09-25 2011-03-31 Xoma Technology Ltd. Screening methods
EP2311873A1 (en) 2004-01-07 2011-04-20 Novartis Vaccines and Diagnostics, Inc. M-CSF-specific monoclonal antibody and uses thereof
WO2011050194A1 (en) 2009-10-22 2011-04-28 Genentech, Inc. Methods and compositions for modulating hepsin activation of macrophage-stimulating protein
WO2011066503A2 (en) 2009-11-30 2011-06-03 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2333069A2 (en) 1998-05-15 2011-06-15 Genentech, Inc. Therapeutic uses of IL-17 homologous polypeptides
EP2335725A1 (en) 2003-04-04 2011-06-22 Genentech, Inc. High concentration antibody and protein formulations
WO2011076922A1 (en) 2009-12-23 2011-06-30 Synimmune Gmbh Anti-flt3 antibodies and methods of using the same
US7982012B2 (en) 2008-03-10 2011-07-19 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of cytomegalovirus
WO2011089211A1 (en) 2010-01-22 2011-07-28 Synimmune Gmbh Anti-cd133 antibodies and methods of using the same
WO2011106297A2 (en) 2010-02-23 2011-09-01 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
WO2011104604A2 (en) 2010-02-23 2011-09-01 Glenmark Pharmaceuticals S.A. Anti-alpha2 integrin antibodies and their uses
EP2368911A1 (en) 2003-05-02 2011-09-28 Xencor Inc. Optimized Fc variants and methods for their generation
US8029783B2 (en) 2005-02-02 2011-10-04 Genentech, Inc. DR5 antibodies and articles of manufacture containing same
EP2383297A1 (en) 2006-08-14 2011-11-02 Xencor Inc. Optimized antibodies that target CD19
WO2011139985A1 (en) 2010-05-03 2011-11-10 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
US8057796B2 (en) 2007-11-12 2011-11-15 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
EP2386640A2 (en) 2004-08-26 2011-11-16 EnGeneIC Molecular Delivery Pty Ltd Delivering functional nucleic acids to mammalian cells via bacterially-derived, intact minicells
US8063187B2 (en) 2007-05-30 2011-11-22 Xencor, Inc. Methods and compositions for inhibiting CD32B expressing cells
WO2011145068A1 (en) 2010-05-20 2011-11-24 Centre National De La Recherche Scientifique Novel self-reactive arms and prodrugs comprising same
EP2389950A1 (en) 2006-03-23 2011-11-30 Novartis AG Anti-tumor cell antigen antibody therapeutics
EP2407483A1 (en) 2006-04-13 2012-01-18 Novartis Vaccines and Diagnostics, Inc. Methods of treating, diagnosing or detecting cancers
EP2423332A1 (en) 2006-08-25 2012-02-29 Oncotherapy Science, Inc. Prognostic markers and therapeutic targets for lung cancer
EP2423231A2 (en) 2006-08-18 2012-02-29 Novartis AG PRLR-specific antibody and uses thereof
EP2447372A2 (en) 2005-07-08 2012-05-02 Biogen Idec MA Inc. Anti alpha v beta 6 antibodies and uses thereof
WO2012061448A1 (en) 2010-11-04 2012-05-10 Boehringer Ingelheim International Gmbh Anti-il-23 antibodies
WO2012085111A1 (en) 2010-12-23 2012-06-28 F. Hoffmann-La Roche Ag Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery
EP2471813A1 (en) 2004-07-15 2012-07-04 Xencor Inc. Optimized Fc variants
EP2474557A2 (en) 2007-07-16 2012-07-11 Genentech, Inc. Anti-CD79b antibodies and immunoconjugates and methods of use
EP2474556A2 (en) 2007-03-14 2012-07-11 Novartis AG APCDD1 inhibitors for treating, diagnosing or detecting cancer
US8236315B2 (en) 2008-01-23 2012-08-07 Glenmark Pharmaceuticals, S.A. Humanized antibodies specific for von Willebrand factor
WO2012109624A2 (en) 2011-02-11 2012-08-16 Zyngenia, Inc. Monovalent and multivalent multispecific complexes and uses thereof
WO2012118813A2 (en) 2011-03-03 2012-09-07 Apexigen, Inc. Anti-il-6 receptor antibodies and methods of use
WO2012122513A2 (en) 2011-03-10 2012-09-13 Omeros Corporation Generation of anti-fn14 monoclonal antibodies by ex-vivo accelerated antibody evolution
US8268970B2 (en) 2007-10-01 2012-09-18 Bristol-Myers Squibb Company Human antibodies that bind mesothelin, and uses thereof
WO2012125614A1 (en) 2011-03-15 2012-09-20 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
US8278421B2 (en) 2006-03-20 2012-10-02 Xoma Techolology Ltd. Human antibodies specific for gastrin materials and methods
US8298531B2 (en) 2008-11-06 2012-10-30 Glenmark Pharmaceuticals, S.A. Treatment with anti-alpha2 integrin antibodies
WO2012149356A2 (en) 2011-04-29 2012-11-01 Apexigen, Inc. Anti-cd40 antibodies and methods of use
WO2012162561A2 (en) 2011-05-24 2012-11-29 Zyngenia, Inc. Multivalent and monovalent multispecific complexes and their uses
EP2532746A2 (en) 2007-03-30 2012-12-12 EnGeneIC Molecular Delivery Pty Ltd. Bacterially-derived, intact minicells that encompass plasmid-free functional nucleic acid for in vivo delivery to mammalian cells
EP2540741A1 (en) 2006-03-06 2013-01-02 Aeres Biomedical Limited Humanized anti-CD22 antibodies and their use in treatment of oncology, transplantation and autoimmune disease
WO2013033069A1 (en) 2011-08-30 2013-03-07 Theraclone Sciences, Inc. Human rhinovirus (hrv) antibodies
WO2013063001A1 (en) 2011-10-28 2013-05-02 Genentech, Inc. Therapeutic combinations and methods of treating melanoma
WO2013074569A1 (en) 2011-11-16 2013-05-23 Boehringer Ingelheim International Gmbh Anti il-36r antibodies
WO2013101771A2 (en) 2011-12-30 2013-07-04 Genentech, Inc. Compositions and method for treating autoimmune diseases
EP2614839A2 (en) 2006-04-05 2013-07-17 Genentech, Inc. Method for using BOC/CDO to modulate hedgehog signaling
WO2013116287A1 (en) 2012-01-31 2013-08-08 Genentech, Inc. Anti-ig-e m1' antibodies and methods using same
WO2013122823A1 (en) 2012-02-13 2013-08-22 Bristol-Myers Squibb Company Enediyne compounds, conjugates thereof, and uses and methods therefor
EP2641618A2 (en) 2007-07-16 2013-09-25 Genentech, Inc. Humanized anti-CD79B antibodies and immunoconjugates and methods of use
WO2013149159A1 (en) 2012-03-30 2013-10-03 Genentech, Inc. Anti-lgr5 antibodies and immunoconjugates
EP2657253A2 (en) 2008-01-31 2013-10-30 Genentech, Inc. Anti-CD79b antibodies and immunoconjugates and methods of use
WO2013165940A1 (en) 2012-05-01 2013-11-07 Genentech, Inc. Anti-pmel17 antibodies and immunoconjugates
WO2013165791A1 (en) 2012-05-03 2013-11-07 Boehringer Ingelheim International Gmbh Anti-il-23p19 antibodies
US8586716B2 (en) 2006-08-04 2013-11-19 Novartis Ag EPHB3-specific antibody and uses thereof
US8591862B2 (en) 2006-06-23 2013-11-26 Engeneic Molecular Delivery Pty Ltd Targeted delivery of drugs, therapeutic nucleic acids and functional nucleic acids to mammalian cells via intact killed bacterial cells
US8609101B2 (en) 2009-04-23 2013-12-17 Theraclone Sciences, Inc. Granulocyte-macrophage colony-stimulating factor (GM-CSF) neutralizing antibodies
USRE44681E1 (en) 2006-07-10 2013-12-31 Biogen Idec Ma Inc. Compositions and methods for inhibiting growth of SMAD4-deficient cancers
US8652469B2 (en) 2005-07-28 2014-02-18 Novartis Ag M-CSF-specific monoclonal antibody and uses thereof
EP2703011A2 (en) 2007-05-07 2014-03-05 MedImmune, LLC Anti-icos antibodies and their use in treatment of oncology, transplantation and autoimmune disease
WO2014047199A1 (en) * 2012-09-19 2014-03-27 The Research Foundation For The State University Of New York Novel prodrugs for selective anticancer therapy
WO2014079886A1 (en) 2012-11-20 2014-05-30 Sanofi Anti-ceacam5 antibodies and uses thereof
WO2014126836A1 (en) 2013-02-14 2014-08-21 Bristol-Myers Squibb Company Tubulysin compounds, methods of making and use
WO2014159835A1 (en) 2013-03-14 2014-10-02 Genentech, Inc. Anti-b7-h4 antibodies and immunoconjugates
US8852586B2 (en) 2004-11-12 2014-10-07 Xencor, Inc. Fc variants with altered binding to FcRn
US8858948B2 (en) 2009-05-20 2014-10-14 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
US8865875B2 (en) 2007-08-22 2014-10-21 Medarex, L.L.C. Site-specific attachment of drugs or other agents to engineered antibodies with C-terminal extensions
EP2796467A1 (en) 2010-03-31 2014-10-29 Boehringer Ingelheim International GmbH Anti-CD40 antibodies
US8900590B2 (en) 2010-08-12 2014-12-02 Theraclone Sciences, Inc. Anti-hemagglutinin antibody compositions and methods of use thereof
US8916160B2 (en) 2011-02-14 2014-12-23 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
US8937158B2 (en) 2003-03-03 2015-01-20 Xencor, Inc. Fc variants with increased affinity for FcγRIIc
WO2015023596A1 (en) 2013-08-12 2015-02-19 Genentech, Inc. Compositions and method for treating complement-associated conditions
WO2015042108A1 (en) 2013-09-17 2015-03-26 Genentech, Inc. Methods of using anti-lgr5 antibodies
US9000132B2 (en) 2013-03-15 2015-04-07 Diadexus, Inc. Lipoprotein-associated phospholipase A2 antibody compositions and methods of use
EP2857516A1 (en) 2000-04-11 2015-04-08 Genentech, Inc. Multivalent antibodies and uses therefor
US9040041B2 (en) 2005-10-03 2015-05-26 Xencor, Inc. Modified FC molecules
WO2015089344A1 (en) 2013-12-13 2015-06-18 Genentech, Inc. Anti-cd33 antibodies and immunoconjugates
WO2015095227A2 (en) 2013-12-16 2015-06-25 Genentech, Inc. Peptidomimetic compounds and antibody-drug conjugates thereof
US9085621B2 (en) 2010-09-10 2015-07-21 Apexigen, Inc. Anti-IL-1β antibodies
WO2015112909A1 (en) 2014-01-24 2015-07-30 Genentech, Inc. Methods of using anti-steap1 antibodies and immunoconjugates
EP2926830A2 (en) 2010-08-31 2015-10-07 Theraclone Sciences, Inc. Human immunodeficiency virus (HIV)-neutralizing antibodies
WO2015179658A2 (en) 2014-05-22 2015-11-26 Genentech, Inc. Anti-gpc3 antibodies and immunoconjugates
US9200079B2 (en) 2004-11-12 2015-12-01 Xencor, Inc. Fc variants with altered binding to FcRn
EP2962697A1 (en) 2006-11-27 2016-01-06 diaDexus, Inc. Ovr110 antibody compositions and methods of use
US9260517B2 (en) 2009-11-17 2016-02-16 Musc Foundation For Research Development Human monoclonal antibodies to human nucleolin
US9278131B2 (en) 2012-08-10 2016-03-08 Adocia Process for lowering the viscosity of highly concentrated protein solutions
WO2016040868A1 (en) 2014-09-12 2016-03-17 Genentech, Inc. Anti-cll-1 antibodies and immunoconjugates
WO2016039801A1 (en) 2014-01-31 2016-03-17 Boehringer Ingelheim International Gmbh Novel anti-baff antibodies
WO2016077260A1 (en) 2014-11-10 2016-05-19 Bristol-Myers Squibb Company Tubulysin analogs and methods of making and use
EP3023497A1 (en) 2005-11-18 2016-05-25 Glenmark Pharmaceuticals S.A. Anti-alpha2 integrin antibodies and their uses
WO2016115201A1 (en) 2015-01-14 2016-07-21 Bristol-Myers Squibb Company Heteroarylene-bridged benzodiazepine dimers, conjugates thereof, and methods of making and using
WO2016141111A1 (en) 2015-03-03 2016-09-09 Xoma (Us) Llc Treatment of post-prandial hyperinsulinemia and hypoglycemia after bariatric surgery
WO2016161410A2 (en) 2015-04-03 2016-10-06 Xoma Technology Ltd. Treatment of cancer using inhibitors of tgf-beta and pd-1
US9465029B2 (en) 2004-04-16 2016-10-11 Glaxo Group Limited Methods for detecting LP-PLA2 activity and inhibition of LP-PLA2 activity
EP3095463A2 (en) 2008-09-16 2016-11-23 F. Hoffmann-La Roche AG Methods for treating progressive multiple sclerosis
WO2016196381A1 (en) 2015-05-29 2016-12-08 Genentech, Inc. Pd-l1 promoter methylation in cancer
EP3112468A1 (en) 1998-05-15 2017-01-04 Genentech, Inc. Il-17 homologous polypeptides and therapeutic uses thereof
US9562099B2 (en) 2013-03-14 2017-02-07 Genentech, Inc. Anti-B7-H4 antibodies and immunoconjugates
EP3130349A1 (en) 2004-06-04 2017-02-15 Genentech, Inc. Method for treating multiple sclerosis
WO2017062682A2 (en) 2015-10-06 2017-04-13 Genentech, Inc. Method for treating multiple sclerosis
US9714282B2 (en) 2003-09-26 2017-07-25 Xencor, Inc. Optimized Fc variants and methods for their generation
EP3208281A1 (en) 2010-03-29 2017-08-23 Zymeworks, Inc. Antibodies with enhanced or suppressed effector function
US9745376B2 (en) 2002-03-13 2017-08-29 Biogen Ma Inc. Anti-ανβ6 antibodies
US9809653B2 (en) 2012-12-27 2017-11-07 Sanofi Anti-LAMP1 antibodies and antibody drug conjugates, and uses thereof
WO2017194568A1 (en) 2016-05-11 2017-11-16 Sanofi Treatment regimen using anti-muc1 maytansinoid immunoconjugate antibody for the treatment of tumors
WO2017214452A1 (en) 2016-06-08 2017-12-14 Xencor, Inc. Treatment of igg4-related diseases with anti-cd19 antibodies crossbinding to cd32b
EP3260136A1 (en) 2009-03-17 2017-12-27 Theraclone Sciences, Inc. Human immunodeficiency virus (hiv) -neutralizing antibodies
WO2018026748A1 (en) 2016-08-01 2018-02-08 Xoma (Us) Llc Parathyroid hormone receptor 1 (pth1r) antibodies and uses thereof
WO2018035391A1 (en) 2016-08-19 2018-02-22 Bristol-Myers Squibb Company Seco-cyclopropapyrroloindole compounds, antibody-drug conjugates thereof, and methods of making and use
WO2018075842A1 (en) 2016-10-20 2018-04-26 Bristol-Myers Squibb Company Condensed benzodiazepine derivatives and conjugates made therefrom
US10035860B2 (en) 2014-03-14 2018-07-31 Biogen Ma Inc. Anti-alpha V beta 6 antibodies and uses thereof

Citations (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4009264A (en) * 1975-03-03 1977-02-22 Meito Sangyo Kabushiki Kaisha Complexes of polysaccharides or derivatives thereof with reduced glutathione and process for preparing said complexes
US4016146A (en) * 1974-12-10 1977-04-05 Biological Developments, Inc. Phenethylamine antigenic conjugates, their preparation, antibodies, and use
US4039519A (en) * 1975-04-14 1977-08-02 Chimie & Biologie Broad spectrum antibiotics and method
US4057685A (en) * 1972-02-02 1977-11-08 Abbott Laboratories Chemically modified endotoxin immunizing agent
US4119589A (en) * 1976-01-29 1978-10-10 Boehringer Mannheim Gmbh Process for fixing proteins onto carriers
US4123519A (en) * 1976-08-17 1978-10-31 Philips Roxane, Inc. Injectable contraceptive vaccine and method
US4140679A (en) * 1976-10-12 1979-02-20 Research Corporation Antigen-protein complex for blocking allergic reactions
US4150105A (en) * 1974-11-29 1979-04-17 Biological Developments, Inc. 3-Ketosteroid antigenic conjugates, their preparation, antibodies and use
US4192799A (en) * 1977-09-21 1980-03-11 The Upjohn Company Conjugates formed by reacting a prostaglandin mimic compound with a carrier molecule
US4193982A (en) * 1975-12-05 1980-03-18 Etablissement Declare D'utilite Publique Dit: Institut Pasteur Process for coupling biological substances by covalent bonds
US4201770A (en) * 1973-05-07 1980-05-06 The Ohio State University Antigenic modification of polypeptides
US4223013A (en) * 1978-12-29 1980-09-16 Syva Company Amitriptyline conjugates to antigenic proteins and enzymes
US4238389A (en) * 1979-02-12 1980-12-09 Syva Company Valproate conjugation using dicarbonyls

Patent Citations (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4057685A (en) * 1972-02-02 1977-11-08 Abbott Laboratories Chemically modified endotoxin immunizing agent
US4201770A (en) * 1973-05-07 1980-05-06 The Ohio State University Antigenic modification of polypeptides
US4150105A (en) * 1974-11-29 1979-04-17 Biological Developments, Inc. 3-Ketosteroid antigenic conjugates, their preparation, antibodies and use
US4016146A (en) * 1974-12-10 1977-04-05 Biological Developments, Inc. Phenethylamine antigenic conjugates, their preparation, antibodies, and use
US4009264A (en) * 1975-03-03 1977-02-22 Meito Sangyo Kabushiki Kaisha Complexes of polysaccharides or derivatives thereof with reduced glutathione and process for preparing said complexes
US4039519A (en) * 1975-04-14 1977-08-02 Chimie & Biologie Broad spectrum antibiotics and method
US4193982A (en) * 1975-12-05 1980-03-18 Etablissement Declare D'utilite Publique Dit: Institut Pasteur Process for coupling biological substances by covalent bonds
US4119589A (en) * 1976-01-29 1978-10-10 Boehringer Mannheim Gmbh Process for fixing proteins onto carriers
US4123519A (en) * 1976-08-17 1978-10-31 Philips Roxane, Inc. Injectable contraceptive vaccine and method
US4140679A (en) * 1976-10-12 1979-02-20 Research Corporation Antigen-protein complex for blocking allergic reactions
US4192799A (en) * 1977-09-21 1980-03-11 The Upjohn Company Conjugates formed by reacting a prostaglandin mimic compound with a carrier molecule
US4223013A (en) * 1978-12-29 1980-09-16 Syva Company Amitriptyline conjugates to antigenic proteins and enzymes
US4238389A (en) * 1979-02-12 1980-12-09 Syva Company Valproate conjugation using dicarbonyls

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP0038357A4 *

Cited By (312)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4912120A (en) * 1986-03-14 1990-03-27 Syntex (U.S.A.) Inc. 3,5-substituted 4,5-dihydroisoxazoles as transglutaminase inhibitors
US4929630A (en) * 1986-03-14 1990-05-29 Syntex (U.S.A.) Inc. Transglutaminase inhibitors
EP0237082A3 (en) * 1986-03-14 1988-09-14 Syntex (U.S.A.) Inc. Transglutaminase inhibitors
US4921941A (en) * 1987-07-01 1990-05-01 Schering Corporation Orally active antiandrogens
US5561119A (en) * 1991-04-30 1996-10-01 Laboratoires Hoechst Glycosylated prodrugs, their method of preparation and their uses
US6051230A (en) * 1992-03-05 2000-04-18 Board Of Regents, The University Of Texas System Compositions for targeting the vasculature of solid tumors
US6451312B1 (en) 1992-03-05 2002-09-17 Board Of Regents, The University Of Texas System VEGF-gelonin for targeting the vasculature of solid tumors
US7112317B2 (en) 1992-03-05 2006-09-26 Board Of Regents, The University Of Texas System Combined methods and compositions for tumor vasculature targeting and tumor treatment
US7691380B2 (en) 1992-03-05 2010-04-06 Board Of Regents, The University Of Texas System Combined methods for tumor coagulation and tumor treatment
US6749853B1 (en) 1992-03-05 2004-06-15 Board Of Regents, The University Of Texas System Combined methods and compositions for coagulation and tumor treatment
US6004554A (en) * 1992-03-05 1999-12-21 Board Of Regents, The University Of Texas System Methods for targeting the vasculature of solid tumors
US5660827A (en) * 1992-03-05 1997-08-26 Board Of Regents, The University Of Texas System Antibodies that bind to endoglin
US5965132A (en) * 1992-03-05 1999-10-12 Board Of Regents, The University Of Texas System Methods and compositions for targeting the vasculature of solid tumors
US5855866A (en) * 1992-03-05 1999-01-05 Board Of Regenis, The University Of Texas System Methods for treating the vasculature of solid tumors
US5863538A (en) * 1992-03-05 1999-01-26 Board Of Regents, The University Of Texas System Compositions for targeting the vasculature of solid tumors
US5877289A (en) * 1992-03-05 1999-03-02 The Scripps Research Institute Tissue factor compositions and ligands for the specific coagulation of vasculature
US7125541B2 (en) 1992-03-05 2006-10-24 The University Of Texas System Board Of Regents Combined methods for tumor vasculature targeting and tumor treatment with radiotherapy
US6004555A (en) * 1992-03-05 1999-12-21 Board Of Regents, The University Of Texas System Methods for the specific coagulation of vasculature
US6093399A (en) * 1992-03-05 2000-07-25 Board Of Regents, The University Of Texas System Methods and compositions for the specific coagulation of vasculature
US6036955A (en) * 1992-03-05 2000-03-14 The Scripps Research Institute Kits and methods for the specific coagulation of vasculature
EP0595133A3 (en) * 1992-10-27 1998-11-04 BEHRINGWERKE Aktiengesellschaft Prodrugs, their preparation and use as medicaments
US6146658A (en) * 1992-10-27 2000-11-14 Hoechst Aktiengesellschaft Prodrugs, their preparation and use as pharmaceuticals
DE4236237A1 (en) * 1992-10-27 1994-04-28 Behringwerke Ag Prodrugs, their preparation and use as medicaments
US5955100A (en) * 1992-10-27 1999-09-21 Behringwerke Aktiengesellschaft Prodrugs their preparation and use as pharmaceuticals
US6214345B1 (en) 1993-05-14 2001-04-10 Bristol-Myers Squibb Co. Lysosomal enzyme-cleavable antitumor drug conjugates
US5621002A (en) * 1993-09-09 1997-04-15 Behringwerke Aktiengesellschaft Prodrugs for enzyme mediated activation
EP0642799A1 (en) * 1993-09-09 1995-03-15 BEHRINGWERKE Aktiengesellschaft Improved prodrugs for enzyme mediated activation
EP0647450A1 (en) * 1993-09-09 1995-04-12 BEHRINGWERKE Aktiengesellschaft Improved prodrugs for enzyme mediated activation
EP0648503A1 (en) * 1993-09-22 1995-04-19 BEHRINGWERKE Aktiengesellschaft Pro-prodrugs, their production and use
US5965566A (en) * 1993-10-20 1999-10-12 Enzon, Inc. High molecular weight polymer-based prodrugs
US6127355A (en) * 1993-10-20 2000-10-03 Enzon, Inc. High molecular weight polymer-based prodrugs
US5880131A (en) * 1993-10-20 1999-03-09 Enzon, Inc. High molecular weight polymer-based prodrugs
WO1996001653A1 (en) * 1994-07-11 1996-01-25 Board Of Regents, The University Of Texas System Methods and compositions for the specific coagulation of vasculature
US7951772B2 (en) 1994-08-19 2011-05-31 La Region Wallonne Tumor-activated prodrug compounds and treatment
US7037898B2 (en) 1994-08-19 2006-05-02 Laregion Wallone Tumor-activated prodrug compounds and treatment
US7390629B2 (en) 1994-08-19 2008-06-24 La Region Wallonne Tumor-activated prodrug compounds and treatment
EP1619250A1 (en) 1996-01-08 2006-01-25 Genentech, Inc. WSX receptor and ligands
EP2301580A1 (en) 1997-04-07 2011-03-30 Genentech, Inc. Anti-VEGF antibodies
EP3260468A1 (en) 1997-04-07 2017-12-27 Genentech, Inc. Anti-vegf antibodies
EP1650220A2 (en) 1997-04-07 2006-04-26 Genentech, Inc. Anti-VEGF antibodies
EP2336190A2 (en) 1997-04-07 2011-06-22 Genentech, Inc. Anti-VEGF antibodies
EP2338915A2 (en) 1997-04-07 2011-06-29 Genentech, Inc. Anti-VEGF antibodies
US6524553B2 (en) 1998-03-31 2003-02-25 Bristol-Myers Squibb Pharma Company Quinolone vitronectin receptor antagonist pharmaceuticals
US6548663B1 (en) 1998-03-31 2003-04-15 Bristol-Myers Squibb Pharma Company Benzodiazepine vitronectin receptor antagonist pharmaceuticals
US6537520B1 (en) 1998-03-31 2003-03-25 Bristol-Myers Squibb Pharma Company Pharmaceuticals for the imaging of angiogenic disorders
US7052673B2 (en) 1998-03-31 2006-05-30 Bristol-Myers Squibb Pharma Company Pharmaceuticals for the imaging of angiogenic disorders
US6395266B1 (en) 1998-04-17 2002-05-28 Enzon, Inc. Terminally-branched polymeric linkers and polymeric conjugates containing the same
US6153655A (en) * 1998-04-17 2000-11-28 Enzon, Inc. Terminally-branched polymeric linkers and polymeric conjugates containing the same
US6638499B2 (en) 1998-04-17 2003-10-28 Enzon, Inc. Terminally-branched polymeric linkers and polymeric conjugates containing the same
EP3112468A1 (en) 1998-05-15 2017-01-04 Genentech, Inc. Il-17 homologous polypeptides and therapeutic uses thereof
EP2333069A2 (en) 1998-05-15 2011-06-15 Genentech, Inc. Therapeutic uses of IL-17 homologous polypeptides
EP2058334A2 (en) 1998-06-12 2009-05-13 Genentech, Inc. Monoclonal antibodies, cross-reactive antibodies and method for producing the same
EP1950300A2 (en) 1998-11-18 2008-07-30 Genentech, Inc. Antibody variants with higher binding affinity compared to parent antibodies
US7018611B2 (en) 1998-12-18 2006-03-28 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6569402B1 (en) 1998-12-18 2003-05-27 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6818201B2 (en) 1998-12-18 2004-11-16 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6794518B1 (en) 1998-12-18 2004-09-21 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US7090828B2 (en) 1998-12-18 2006-08-15 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6743412B2 (en) 1998-12-18 2004-06-01 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6689337B2 (en) 1998-12-18 2004-02-10 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6558649B1 (en) 1998-12-18 2003-05-06 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US7332149B1 (en) 1998-12-18 2008-02-19 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6511649B1 (en) 1998-12-18 2003-01-28 Thomas D. Harris Vitronectin receptor antagonist pharmaceuticals
US7321045B2 (en) 1998-12-18 2008-01-22 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6683163B2 (en) 1998-12-18 2004-01-27 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
US6511648B2 (en) 1998-12-18 2003-01-28 Bristol-Myers Squibb Pharma Company Vitronectin receptor antagonist pharmaceuticals
EP2112167A2 (en) 1999-06-25 2009-10-28 Genentech, Inc. Humanized ANTI-ERBB2 antibodies and treatment with ANTI-ERBB2 antibodies
EP2111870A1 (en) 1999-08-27 2009-10-28 Genentech, Inc. Dosages for treatment of anti-erbB2 antibodies
EP2110138A1 (en) 1999-08-27 2009-10-21 Genentech, Inc. Dosages for treatment of anti-erbB2 antibodies
US7855278B2 (en) 2000-01-13 2010-12-21 Genentech, Inc. Antibodies to Stra6 polypeptides
US7173115B2 (en) 2000-01-13 2007-02-06 Genentech, Inc. Stra6 polypeptides
US7939650B2 (en) 2000-01-13 2011-05-10 Genentech, Inc. Stra6 polypeptides
US7741439B2 (en) 2000-01-13 2010-06-22 Genentech, Inc. Isolated stra6 polypeptides
EP2857516A1 (en) 2000-04-11 2015-04-08 Genentech, Inc. Multivalent antibodies and uses therefor
US7696313B2 (en) 2000-06-14 2010-04-13 Medarex, Inc. Prodrug compounds with isoleucine
US7329507B2 (en) 2000-06-14 2008-02-12 Medarex, Inc. Prodrug compounds with isoleucine
US7115573B2 (en) 2000-06-14 2006-10-03 Medarex, Inc. Prodrug compounds with an isoleucine
EP2052742A1 (en) 2000-06-20 2009-04-29 Biogen Idec Inc. Treatment of B-cell associated diseases such as malignancies and autoimmune diseases using a cold anti-CD20 antibody/radiolabeled anti-CD22 antibody combination
EP2014303A2 (en) 2000-07-27 2009-01-14 Genentech, Inc. APO-2L receptor agonist and CPT-11 synergism
WO2002022833A1 (en) * 2000-09-15 2002-03-21 Universität Stuttgart Fusion protein from antibody cytokine-cytokine inhibitor (selectokine) for use as target-specific prodrug
WO2002030463A2 (en) 2000-10-12 2002-04-18 Genentech, Inc. Reduced-viscosity concentrated protein formulations
EP2292301A2 (en) 2001-02-22 2011-03-09 Genentech, Inc. Anti-interferon-alpha antibodies
EP2065467A2 (en) 2001-02-22 2009-06-03 Genentech, Inc. Anti-interferon-alpha antibodies
EP1243276A1 (en) * 2001-03-23 2002-09-25 Patrick Henry Beusker Elongated and multiple spacers containing activatible prodrugs
US7223837B2 (en) 2001-03-23 2007-05-29 Syntarga B.V. Elongated and multiple spacers in activatible prodrugs
WO2002083180A1 (en) * 2001-03-23 2002-10-24 Syntarga B.V. Elongated and multiple spacers in activatible prodrugs
EP1992643A2 (en) 2001-06-20 2008-11-19 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2000148A1 (en) 2001-06-20 2008-12-10 Genentech, Inc. Compositions and methods for the diagnosis and treatment of prostate cancer
EP2000545A1 (en) 2001-06-20 2008-12-10 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2000482A1 (en) 2001-06-20 2008-12-10 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2151244A1 (en) 2001-09-18 2010-02-10 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2153843A1 (en) 2001-09-18 2010-02-17 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2143438A1 (en) 2001-09-18 2010-01-13 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2067472A1 (en) 2002-01-02 2009-06-10 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
US9745376B2 (en) 2002-03-13 2017-08-29 Biogen Ma Inc. Anti-ανβ6 antibodies
EP2011886A2 (en) 2002-04-16 2009-01-07 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP2263691A1 (en) 2002-07-15 2010-12-22 Genentech, Inc. Treatment of cancer with the recombinant humanized monoclonal anti-erbb2 antibody 2C4 (rhuMAb 2C4)
EP2298805A2 (en) 2002-09-27 2011-03-23 Xencor, Inc. Optimized Fc variants and methods for their generation
EP3150630A1 (en) 2002-09-27 2017-04-05 Xencor Inc. Optimized fc variants and methods for their generation
EP2345671A1 (en) 2002-09-27 2011-07-20 Xencor Inc. Optimized fc variants and methods for their generation
EP2364996A1 (en) 2002-09-27 2011-09-14 Xencor Inc. Optimized FC variants and methods for their generation
EP3321282A1 (en) 2002-09-27 2018-05-16 Xencor, Inc. Optimized fc variants and methods for their generation
EP2042517A1 (en) 2002-09-27 2009-04-01 Xencor, Inc. Optimized FC variants and methods for their generation
US8937158B2 (en) 2003-03-03 2015-01-20 Xencor, Inc. Fc variants with increased affinity for FcγRIIc
EP2335725A1 (en) 2003-04-04 2011-06-22 Genentech, Inc. High concentration antibody and protein formulations
EP3178492A1 (en) 2003-04-04 2017-06-14 Genentech, Inc. High concentration antibody and protein formulations
EP2062916A2 (en) 2003-04-09 2009-05-27 Genentech, Inc. Therapy of autoimmune disease in a patient with an inadequate response to a TNF-Alpha inhibitor
EP2368911A1 (en) 2003-05-02 2011-09-28 Xencor Inc. Optimized Fc variants and methods for their generation
EP3101030A1 (en) 2003-05-02 2016-12-07 Xencor, Inc. Optimized fc variants and methods for their generation
EP2311875A1 (en) 2003-05-30 2011-04-20 Genentech, Inc. Treatment with anti-VEGF antibodies
EP2251355A1 (en) 2003-05-30 2010-11-17 Genentech, Inc. Treatment with anti-VEGF antibodies
EP2248829A1 (en) 2003-05-30 2010-11-10 Genentech, Inc. Treatment with anti-VEGF antibodies
US9714282B2 (en) 2003-09-26 2017-07-25 Xencor, Inc. Optimized Fc variants and methods for their generation
EP2301568A1 (en) 2003-11-17 2011-03-30 Genentech, Inc. Antibody against IRTA2 for the treatment of tumour of hematopoietic origin
EP2295073A1 (en) 2003-11-17 2011-03-16 Genentech, Inc. Antibody against CD22 for the treatment of tumour of hematopoietic origin
EP2161283A1 (en) 2003-11-17 2010-03-10 Genentech, Inc. Compositions comprising antibodies against CD79b conjugated to a growth inhibitory agent or cytotoxic agent and methods for the treatment of tumor of hematopoietic origin
EP2311873A1 (en) 2004-01-07 2011-04-20 Novartis Vaccines and Diagnostics, Inc. M-CSF-specific monoclonal antibody and uses thereof
US9465029B2 (en) 2004-04-16 2016-10-11 Glaxo Group Limited Methods for detecting LP-PLA2 activity and inhibition of LP-PLA2 activity
US7655808B2 (en) 2004-06-03 2010-02-02 Bristol-Myers Squibb Company Leptomycin compounds
US7446196B2 (en) 2004-06-03 2008-11-04 Kosan Biosciences, Incorporated Leptomycin compounds
EP3130349A1 (en) 2004-06-04 2017-02-15 Genentech, Inc. Method for treating multiple sclerosis
US7541330B2 (en) 2004-06-15 2009-06-02 Kosan Biosciences Incorporated Conjugates with reduced adverse systemic effects
WO2006009901A2 (en) 2004-06-18 2006-01-26 Ambrx, Inc. Novel antigen-binding polypeptides and their uses
EP2471813A1 (en) 2004-07-15 2012-07-04 Xencor Inc. Optimized Fc variants
EP2386640A2 (en) 2004-08-26 2011-11-16 EnGeneIC Molecular Delivery Pty Ltd Delivering functional nucleic acids to mammalian cells via bacterially-derived, intact minicells
US9242007B2 (en) 2004-08-26 2016-01-26 Engeneic Molecular Delivery Pty. Ltd. Delivering functional nucleic acids to mammalian cells via bacterially-derived, intact minicells
US8735566B2 (en) 2004-08-26 2014-05-27 Engeneic Molecular Delivery Pty Ltd Bacterially derived intact minicells that encompass plasmid free functional nucleic acid for in vivo delivery to mammalian cells
US9730897B2 (en) 2004-08-26 2017-08-15 Engeneic Molecular Delivery Pty Ltd Delivering functional nucleic acids to mammalian cells via bacterially-derived, intact minicells
US8691963B2 (en) 2004-08-26 2014-04-08 Engeneic Molecular Delivery Pty. Ltd. Delivering functional nucleic acids to mammalian cells via bacterially-derived, intact minicells
US8956864B2 (en) 2004-08-26 2015-02-17 Engeneic Molecular Delivery Pty Ltd Bacterially derived intact minicells that encompass plasmid free functional nucleic acid for in vivo delivery to mammalian cells
US8669101B2 (en) 2004-08-26 2014-03-11 Engeneic Molecular Delivery Pty. Ltd. Bacterially derived intact minicells that encompass plasmid free functional nucleic acid for in vivo delivery to mammalian cells
US9066982B2 (en) 2004-08-26 2015-06-30 Engeneic Molecular Delivery Pty Ltd. Bacterially-derived, intact minicells that encompass plasmid-free functional nucleic acid for in vivo delivery to mammalian cells
US9200079B2 (en) 2004-11-12 2015-12-01 Xencor, Inc. Fc variants with altered binding to FcRn
US9803023B2 (en) 2004-11-12 2017-10-31 Xencor, Inc. Fc variants with altered binding to FcRn
US8852586B2 (en) 2004-11-12 2014-10-07 Xencor, Inc. Fc variants with altered binding to FcRn
US8883973B2 (en) 2004-11-12 2014-11-11 Xencor, Inc. Fc variants with altered binding to FcRn
EP2230517A1 (en) 2005-01-07 2010-09-22 Diadexus, Inc. OVR110 antibody compositions and methods of use
WO2006074418A2 (en) 2005-01-07 2006-07-13 Diadexus, Inc. Ovr110 antibody compositions and methods of use
US8030023B2 (en) 2005-02-02 2011-10-04 Genentech, Inc. Nucleic acid encoding DR5 antibodies and uses thereof
US8409570B2 (en) 2005-02-02 2013-04-02 Genentech, Inc. Method of inducing apoptosis using anti-DR5 antibodies
US8029783B2 (en) 2005-02-02 2011-10-04 Genentech, Inc. DR5 antibodies and articles of manufacture containing same
WO2006089133A2 (en) 2005-02-15 2006-08-24 Duke University Anti-cd19 antibodies and uses in oncology
EP2548575A1 (en) 2005-02-15 2013-01-23 Duke University Anti-CD19 antibodies that mediate ADCC for use in treating autoimmune diseases
EP2062591A1 (en) 2005-04-07 2009-05-27 Novartis Vaccines and Diagnostics, Inc. CACNA1E in cancer diagnosis detection and treatment
EP2083088A2 (en) 2005-04-07 2009-07-29 Novartis Vaccines and Diagnostics, Inc. Cancer-related genes
EP2221316A1 (en) 2005-05-05 2010-08-25 Duke University Anti-CD19 antibody therapy for autoimmune disease
EP2447372A2 (en) 2005-07-08 2012-05-02 Biogen Idec MA Inc. Anti alpha v beta 6 antibodies and uses thereof
US8992924B2 (en) 2005-07-08 2015-03-31 Biogen Idec Ma Inc. Anti-ανβ6 antibodies and uses thereof
EP2458012A1 (en) 2005-07-08 2012-05-30 Biogen Idec MA Inc. Anti alpha v beta 6 antibodies and uses thereof
EP2458013A1 (en) 2005-07-08 2012-05-30 Biogen Idec MA Inc. Anti alpha v beta 6 antibodies and uses therof
US8652469B2 (en) 2005-07-28 2014-02-18 Novartis Ag M-CSF-specific monoclonal antibody and uses thereof
US9040041B2 (en) 2005-10-03 2015-05-26 Xencor, Inc. Modified FC molecules
EP3023497A1 (en) 2005-11-18 2016-05-25 Glenmark Pharmaceuticals S.A. Anti-alpha2 integrin antibodies and their uses
EP2540741A1 (en) 2006-03-06 2013-01-02 Aeres Biomedical Limited Humanized anti-CD22 antibodies and their use in treatment of oncology, transplantation and autoimmune disease
US8278421B2 (en) 2006-03-20 2012-10-02 Xoma Techolology Ltd. Human antibodies specific for gastrin materials and methods
EP2389950A1 (en) 2006-03-23 2011-11-30 Novartis AG Anti-tumor cell antigen antibody therapeutics
EP2389951A1 (en) 2006-03-23 2011-11-30 Novartis AG Anti-tumor cell antigen antibody therapeutics
EP2389946A1 (en) 2006-03-23 2011-11-30 Novartis AG Anti-tumor cell antigen antibody therapeutics
EP2389949A1 (en) 2006-03-23 2011-11-30 Novartis AG Anti-tumor cell antigen antibody therapeutics
EP2389948A1 (en) 2006-03-23 2011-11-30 Novartis AG Anti-tumor cell antigen antibody therapeutics
EP2389947A1 (en) 2006-03-23 2011-11-30 Novartis AG Anti-tumor cell antigen antibody therapeutics
EP2614839A2 (en) 2006-04-05 2013-07-17 Genentech, Inc. Method for using BOC/CDO to modulate hedgehog signaling
EP2407483A1 (en) 2006-04-13 2012-01-18 Novartis Vaccines and Diagnostics, Inc. Methods of treating, diagnosing or detecting cancers
US8591862B2 (en) 2006-06-23 2013-11-26 Engeneic Molecular Delivery Pty Ltd Targeted delivery of drugs, therapeutic nucleic acids and functional nucleic acids to mammalian cells via intact killed bacterial cells
EP2712618A1 (en) 2006-06-23 2014-04-02 EnGeneIC Molecular Delivery Pty Ltd. Targeted delivery of drugs, therapeutic nucleic acids and functional nucleic acids to mammalian cells via intact killed bacterial cells
US9878043B2 (en) 2006-06-23 2018-01-30 Engeneic Molecular Delivery Pty Ltd Targeted delivery of drugs, therapeutic nucleic acids and functional nucleic acids to mammalian cells via intact killed bacterial cells
USRE44681E1 (en) 2006-07-10 2013-12-31 Biogen Idec Ma Inc. Compositions and methods for inhibiting growth of SMAD4-deficient cancers
WO2008011081A2 (en) 2006-07-19 2008-01-24 The Trustees Of The University Of Pennsylvania Wsx-1/p28 as a target for anti-inflammatory responses
US8586716B2 (en) 2006-08-04 2013-11-19 Novartis Ag EPHB3-specific antibody and uses thereof
US9006398B2 (en) 2006-08-04 2015-04-14 Novartis Ag EPHB3-specific antibody and uses thereof
EP2383297A1 (en) 2006-08-14 2011-11-02 Xencor Inc. Optimized antibodies that target CD19
EP2423231A2 (en) 2006-08-18 2012-02-29 Novartis AG PRLR-specific antibody and uses thereof
EP3018144A1 (en) 2006-08-18 2016-05-11 XOMA Technology Ltd. Prlr-specific antibody and uses thereof
US9005614B2 (en) 2006-08-18 2015-04-14 Novartis Ag PRLR-specific antibody and uses thereof
EP2423332A1 (en) 2006-08-25 2012-02-29 Oncotherapy Science, Inc. Prognostic markers and therapeutic targets for lung cancer
EP2423333A1 (en) 2006-08-25 2012-02-29 Oncotherapy Science, Inc. Prognostic markers and therapeutic targets for lung cancer
EP2962697A1 (en) 2006-11-27 2016-01-06 diaDexus, Inc. Ovr110 antibody compositions and methods of use
EP2474556A2 (en) 2007-03-14 2012-07-11 Novartis AG APCDD1 inhibitors for treating, diagnosing or detecting cancer
EP2532746A2 (en) 2007-03-30 2012-12-12 EnGeneIC Molecular Delivery Pty Ltd. Bacterially-derived, intact minicells that encompass plasmid-free functional nucleic acid for in vivo delivery to mammalian cells
EP2865755A2 (en) 2007-03-30 2015-04-29 EnGeneIC Molecular Delivery Pty Ltd Bacterially derived, intact minicells encompassing regulatory RNA
EP3205724A1 (en) 2007-03-30 2017-08-16 EnGeneIC Molecular Delivery Pty Ltd. Bacterially derived, intact minicells encompassing regulatory rna
EP2703011A2 (en) 2007-05-07 2014-03-05 MedImmune, LLC Anti-icos antibodies and their use in treatment of oncology, transplantation and autoimmune disease
EP2737907A2 (en) 2007-05-07 2014-06-04 MedImmune, LLC Anti-icos antibodies and their use in treatment of oncology, transplantation and autoimmune disease
US9260523B2 (en) 2007-05-30 2016-02-16 Xencor, Inc. Methods and compositions for inhibiting CD32b expressing cells
US9914778B2 (en) 2007-05-30 2018-03-13 Xencor, Inc. Methods and compositions for inhibiting CD32B expressing cells
US9079960B2 (en) 2007-05-30 2015-07-14 Xencor, Inc. Methods and compositions for inhibiting CD32B expressing cells
US9902773B2 (en) 2007-05-30 2018-02-27 Xencor, Inc. Methods and compositions for inhibiting CD32b expressing cells
US8063187B2 (en) 2007-05-30 2011-11-22 Xencor, Inc. Methods and compositions for inhibiting CD32B expressing cells
US9394366B2 (en) 2007-05-30 2016-07-19 Xencor, Inc. Methods and compositions for inhibiting CD32B expressing cells
EP2708557A1 (en) 2007-05-30 2014-03-19 Xencor, Inc. Method and compositions for inhibiting CD32B expressing cells
EP2474557A2 (en) 2007-07-16 2012-07-11 Genentech, Inc. Anti-CD79b antibodies and immunoconjugates and methods of use
EP2641618A2 (en) 2007-07-16 2013-09-25 Genentech, Inc. Humanized anti-CD79B antibodies and immunoconjugates and methods of use
EP2502937A2 (en) 2007-07-16 2012-09-26 Genentech, Inc. Anti-CD 79b Antibodies And Immunoconjugates And Methods Of Use
US8865875B2 (en) 2007-08-22 2014-10-21 Medarex, L.L.C. Site-specific attachment of drugs or other agents to engineered antibodies with C-terminal extensions
WO2009028158A1 (en) 2007-08-24 2009-03-05 Oncotherapy Science, Inc. Dkk1 oncogene as therapeutic target for cancer and a diagnosing marker
US8268970B2 (en) 2007-10-01 2012-09-18 Bristol-Myers Squibb Company Human antibodies that bind mesothelin, and uses thereof
US8425904B2 (en) 2007-10-01 2013-04-23 Bristol-Myers Squibb Company Human antibodies that bind mesothelin, and uses thereof
US8383779B2 (en) 2007-10-01 2013-02-26 Bristol-Myers Squibb Company Human antibodies that bind mesothelin, and uses thereof
US8399623B2 (en) 2007-10-01 2013-03-19 Bristol-Myers Squibb Company Human antibodies that bind mesothelin, and uses thereof
US8114402B2 (en) 2007-11-12 2012-02-14 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
US8057796B2 (en) 2007-11-12 2011-11-15 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
US8460671B2 (en) 2007-11-12 2013-06-11 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
US8236315B2 (en) 2008-01-23 2012-08-07 Glenmark Pharmaceuticals, S.A. Humanized antibodies specific for von Willebrand factor
EP2657253A2 (en) 2008-01-31 2013-10-30 Genentech, Inc. Anti-CD79b antibodies and immunoconjugates and methods of use
US7982012B2 (en) 2008-03-10 2011-07-19 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of cytomegalovirus
US8268309B2 (en) 2008-03-10 2012-09-18 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of cytomegalovirus
US8852594B2 (en) 2008-03-10 2014-10-07 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of cytomegalovirus infections
WO2010001585A1 (en) 2008-06-30 2010-01-07 Oncotherapy Science, Inc. Anti-cdh3 antibodies labeled with radioisotope label and uses thereof
US9994642B2 (en) 2008-09-16 2018-06-12 Genentech, Inc. Methods for treating progressive multiple sclerosis
US9683047B2 (en) 2008-09-16 2017-06-20 Genentech, Inc. Methods for treating progressive multiple sclerosis
EP3095463A2 (en) 2008-09-16 2016-11-23 F. Hoffmann-La Roche AG Methods for treating progressive multiple sclerosis
US8298531B2 (en) 2008-11-06 2012-10-30 Glenmark Pharmaceuticals, S.A. Treatment with anti-alpha2 integrin antibodies
WO2010056804A1 (en) 2008-11-12 2010-05-20 Medimmune, Llc Antibody formulation
WO2010060920A1 (en) 2008-11-27 2010-06-03 INSERM (Institut National de la Santé et de la Recherche Médicale) Cxcl4l1 as a biomarker of pancreatic cancer
EP3255060A1 (en) 2008-12-09 2017-12-13 F. Hoffmann-La Roche AG Anti-pd-l1 antibodies and their use to enhance t-cell function
WO2010077634A1 (en) 2008-12-09 2010-07-08 Genentech, Inc. Anti-pd-l1 antibodies and their use to enhance t-cell function
WO2010102175A1 (en) 2009-03-05 2010-09-10 Medarex, Inc. Fully human antibodies specific to cadm1
EP3260136A1 (en) 2009-03-17 2017-12-27 Theraclone Sciences, Inc. Human immunodeficiency virus (hiv) -neutralizing antibodies
EP3323427A1 (en) 2009-03-17 2018-05-23 Theraclone Sciences, Inc. Human immunodeficiency virus (hiv)-neutralizing antibodies
WO2010120561A1 (en) 2009-04-01 2010-10-21 Genentech, Inc. Anti-fcrh5 antibodies and immunoconjugates and methods of use
US8609101B2 (en) 2009-04-23 2013-12-17 Theraclone Sciences, Inc. Granulocyte-macrophage colony-stimulating factor (GM-CSF) neutralizing antibodies
US8858948B2 (en) 2009-05-20 2014-10-14 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
WO2011017249A1 (en) 2009-08-03 2011-02-10 Medarex, Inc. Antiproliferative compounds, conjugates thereof, methods therefor, and uses thereof
EP3309168A1 (en) 2009-08-06 2018-04-18 F. Hoffmann-La Roche AG Method to improve virus removal in protein purification
WO2011031397A1 (en) 2009-08-06 2011-03-17 Genentech, Inc. Method to improve virus removal in protein purification
EP2957296A1 (en) 2009-09-25 2015-12-23 Xoma (Us) Llc Insulin receptor binding antibodies
WO2011038302A2 (en) 2009-09-25 2011-03-31 Xoma Technology Ltd. Novel modulators
EP3187877A1 (en) 2009-09-25 2017-07-05 XOMA Technology Ltd. Screening methods
WO2011038301A2 (en) 2009-09-25 2011-03-31 Xoma Technology Ltd. Screening methods
WO2011050194A1 (en) 2009-10-22 2011-04-28 Genentech, Inc. Methods and compositions for modulating hepsin activation of macrophage-stimulating protein
US8697386B2 (en) 2009-10-22 2014-04-15 Genentech, Inc. Methods and compositions for modulating hepsin activation of macrophage-stimulating protein
US9260517B2 (en) 2009-11-17 2016-02-16 Musc Foundation For Research Development Human monoclonal antibodies to human nucleolin
EP3037435A1 (en) 2009-11-17 2016-06-29 MUSC Foundation For Research Development Human monoclonal antibodies to human nucleolin
EP3002297A2 (en) 2009-11-30 2016-04-06 F. Hoffmann-La Roche AG Antibodies for treating and diagnosing tumors expressing slc34a2 (tat211)
WO2011066503A2 (en) 2009-11-30 2011-06-03 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
WO2011076922A1 (en) 2009-12-23 2011-06-30 Synimmune Gmbh Anti-flt3 antibodies and methods of using the same
WO2011089211A1 (en) 2010-01-22 2011-07-28 Synimmune Gmbh Anti-cd133 antibodies and methods of using the same
WO2011106297A2 (en) 2010-02-23 2011-09-01 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
WO2011104604A2 (en) 2010-02-23 2011-09-01 Glenmark Pharmaceuticals S.A. Anti-alpha2 integrin antibodies and their uses
EP2848632A1 (en) 2010-02-23 2015-03-18 Sanofi Anti-alpha2 integrin antibodies and their uses
EP3208281A1 (en) 2010-03-29 2017-08-23 Zymeworks, Inc. Antibodies with enhanced or suppressed effector function
EP3178851A1 (en) 2010-03-31 2017-06-14 Boehringer Ingelheim International GmbH Anti-cd40 antibodies
EP2796467A1 (en) 2010-03-31 2014-10-29 Boehringer Ingelheim International GmbH Anti-CD40 antibodies
WO2011139985A1 (en) 2010-05-03 2011-11-10 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
WO2011145068A1 (en) 2010-05-20 2011-11-24 Centre National De La Recherche Scientifique Novel self-reactive arms and prodrugs comprising same
US9000135B2 (en) 2010-05-20 2015-04-07 Centre Nationale De Recherche Scientifique Self-reactive arms and prodrugs comprising same
US8900590B2 (en) 2010-08-12 2014-12-02 Theraclone Sciences, Inc. Anti-hemagglutinin antibody compositions and methods of use thereof
EP2926830A2 (en) 2010-08-31 2015-10-07 Theraclone Sciences, Inc. Human immunodeficiency virus (HIV)-neutralizing antibodies
US9085621B2 (en) 2010-09-10 2015-07-21 Apexigen, Inc. Anti-IL-1β antibodies
EP3281954A1 (en) 2010-11-04 2018-02-14 Boehringer Ingelheim International GmbH Anti-il-23 antibodies
WO2012061448A1 (en) 2010-11-04 2012-05-10 Boehringer Ingelheim International Gmbh Anti-il-23 antibodies
EP3115375A1 (en) 2010-11-04 2017-01-11 Boehringer Ingelheim International GmbH Anti-il-23 antibodies
WO2012085111A1 (en) 2010-12-23 2012-06-28 F. Hoffmann-La Roche Ag Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery
WO2012109624A2 (en) 2011-02-11 2012-08-16 Zyngenia, Inc. Monovalent and multivalent multispecific complexes and uses thereof
US8916160B2 (en) 2011-02-14 2014-12-23 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
WO2012118813A2 (en) 2011-03-03 2012-09-07 Apexigen, Inc. Anti-il-6 receptor antibodies and methods of use
WO2012122513A2 (en) 2011-03-10 2012-09-13 Omeros Corporation Generation of anti-fn14 monoclonal antibodies by ex-vivo accelerated antibody evolution
WO2012125614A1 (en) 2011-03-15 2012-09-20 Theraclone Sciences, Inc. Compositions and methods for the therapy and diagnosis of influenza
WO2012149356A2 (en) 2011-04-29 2012-11-01 Apexigen, Inc. Anti-cd40 antibodies and methods of use
WO2012162561A2 (en) 2011-05-24 2012-11-29 Zyngenia, Inc. Multivalent and monovalent multispecific complexes and their uses
US8822651B2 (en) 2011-08-30 2014-09-02 Theraclone Sciences, Inc. Human rhinovirus (HRV) antibodies
WO2013033069A1 (en) 2011-08-30 2013-03-07 Theraclone Sciences, Inc. Human rhinovirus (hrv) antibodies
WO2013063001A1 (en) 2011-10-28 2013-05-02 Genentech, Inc. Therapeutic combinations and methods of treating melanoma
WO2013074569A1 (en) 2011-11-16 2013-05-23 Boehringer Ingelheim International Gmbh Anti il-36r antibodies
WO2013101771A2 (en) 2011-12-30 2013-07-04 Genentech, Inc. Compositions and method for treating autoimmune diseases
WO2013116287A1 (en) 2012-01-31 2013-08-08 Genentech, Inc. Anti-ig-e m1' antibodies and methods using same
US9156850B2 (en) 2012-02-13 2015-10-13 Bristol-Myers Squibb Company Enediyne compounds, conjugates thereof, and uses and methods therefor
US8709431B2 (en) 2012-02-13 2014-04-29 Bristol-Myers Squibb Company Enediyne compounds, conjugates thereof, and uses and methods therefor
WO2013122823A1 (en) 2012-02-13 2013-08-22 Bristol-Myers Squibb Company Enediyne compounds, conjugates thereof, and uses and methods therefor
WO2013149159A1 (en) 2012-03-30 2013-10-03 Genentech, Inc. Anti-lgr5 antibodies and immunoconjugates
US9175089B2 (en) 2012-03-30 2015-11-03 Genentech, Inc. Anti-LGR5 antibodies and immunoconjugates
WO2013165940A1 (en) 2012-05-01 2013-11-07 Genentech, Inc. Anti-pmel17 antibodies and immunoconjugates
US9056910B2 (en) 2012-05-01 2015-06-16 Genentech, Inc. Anti-PMEL17 antibodies and immunoconjugates
US9597411B2 (en) 2012-05-01 2017-03-21 Genentech, Inc. Anti-PMEL17 antibodies and immunoconjugates
EP3326649A1 (en) 2012-05-03 2018-05-30 Boehringer Ingelheim International GmbH Anti-il-23p19 antibodies
WO2013165791A1 (en) 2012-05-03 2013-11-07 Boehringer Ingelheim International Gmbh Anti-il-23p19 antibodies
US9278131B2 (en) 2012-08-10 2016-03-08 Adocia Process for lowering the viscosity of highly concentrated protein solutions
US9872919B2 (en) 2012-09-19 2018-01-23 The Research Foundation For The State University Of New York Prodrugs for selective anticancer therapy
WO2014047199A1 (en) * 2012-09-19 2014-03-27 The Research Foundation For The State University Of New York Novel prodrugs for selective anticancer therapy
US9617345B2 (en) 2012-11-20 2017-04-11 Sanofi Anti-CEACAM5 antibodies and uses thereof
WO2014079886A1 (en) 2012-11-20 2014-05-30 Sanofi Anti-ceacam5 antibodies and uses thereof
US9809653B2 (en) 2012-12-27 2017-11-07 Sanofi Anti-LAMP1 antibodies and antibody drug conjugates, and uses thereof
US9109008B2 (en) 2013-02-14 2015-08-18 Bristol-Myers Squibb Company Tubulysin compounds, methods of making and use
US8980824B2 (en) 2013-02-14 2015-03-17 Bristol-Myers Squibb Company Tubulysin compounds, methods of making and use
US9688721B2 (en) 2013-02-14 2017-06-27 Bristol-Myers Squibb Company Tubulysin compounds, methods of making and use
WO2014126836A1 (en) 2013-02-14 2014-08-21 Bristol-Myers Squibb Company Tubulysin compounds, methods of making and use
US9382289B2 (en) 2013-02-14 2016-07-05 Bristol-Myers Squibb Company Tubulysin compounds, methods of making and use
WO2014159835A1 (en) 2013-03-14 2014-10-02 Genentech, Inc. Anti-b7-h4 antibodies and immunoconjugates
US9562099B2 (en) 2013-03-14 2017-02-07 Genentech, Inc. Anti-B7-H4 antibodies and immunoconjugates
EP3299391A1 (en) 2013-03-14 2018-03-28 Genentech, Inc. Anti-b7-h4 antibodies and immunoconjugates
US9000132B2 (en) 2013-03-15 2015-04-07 Diadexus, Inc. Lipoprotein-associated phospholipase A2 antibody compositions and methods of use
WO2015023596A1 (en) 2013-08-12 2015-02-19 Genentech, Inc. Compositions and method for treating complement-associated conditions
WO2015042108A1 (en) 2013-09-17 2015-03-26 Genentech, Inc. Methods of using anti-lgr5 antibodies
WO2015089344A1 (en) 2013-12-13 2015-06-18 Genentech, Inc. Anti-cd33 antibodies and immunoconjugates
WO2015095227A2 (en) 2013-12-16 2015-06-25 Genentech, Inc. Peptidomimetic compounds and antibody-drug conjugates thereof
WO2015112909A1 (en) 2014-01-24 2015-07-30 Genentech, Inc. Methods of using anti-steap1 antibodies and immunoconjugates
WO2016039801A1 (en) 2014-01-31 2016-03-17 Boehringer Ingelheim International Gmbh Novel anti-baff antibodies
US10035859B2 (en) 2014-03-14 2018-07-31 Biogen Ma Inc. Anti-alpha V beta 6 antibodies and uses thereof
US10035860B2 (en) 2014-03-14 2018-07-31 Biogen Ma Inc. Anti-alpha V beta 6 antibodies and uses thereof
WO2015179658A2 (en) 2014-05-22 2015-11-26 Genentech, Inc. Anti-gpc3 antibodies and immunoconjugates
WO2016040868A1 (en) 2014-09-12 2016-03-17 Genentech, Inc. Anti-cll-1 antibodies and immunoconjugates
WO2016077260A1 (en) 2014-11-10 2016-05-19 Bristol-Myers Squibb Company Tubulysin analogs and methods of making and use
WO2016115201A1 (en) 2015-01-14 2016-07-21 Bristol-Myers Squibb Company Heteroarylene-bridged benzodiazepine dimers, conjugates thereof, and methods of making and using
WO2016141111A1 (en) 2015-03-03 2016-09-09 Xoma (Us) Llc Treatment of post-prandial hyperinsulinemia and hypoglycemia after bariatric surgery
WO2016161410A2 (en) 2015-04-03 2016-10-06 Xoma Technology Ltd. Treatment of cancer using inhibitors of tgf-beta and pd-1
WO2016196381A1 (en) 2015-05-29 2016-12-08 Genentech, Inc. Pd-l1 promoter methylation in cancer
WO2017062682A2 (en) 2015-10-06 2017-04-13 Genentech, Inc. Method for treating multiple sclerosis
WO2017194568A1 (en) 2016-05-11 2017-11-16 Sanofi Treatment regimen using anti-muc1 maytansinoid immunoconjugate antibody for the treatment of tumors
WO2017214452A1 (en) 2016-06-08 2017-12-14 Xencor, Inc. Treatment of igg4-related diseases with anti-cd19 antibodies crossbinding to cd32b
WO2018026748A1 (en) 2016-08-01 2018-02-08 Xoma (Us) Llc Parathyroid hormone receptor 1 (pth1r) antibodies and uses thereof
WO2018035391A1 (en) 2016-08-19 2018-02-22 Bristol-Myers Squibb Company Seco-cyclopropapyrroloindole compounds, antibody-drug conjugates thereof, and methods of making and use
WO2018075842A1 (en) 2016-10-20 2018-04-26 Bristol-Myers Squibb Company Condensed benzodiazepine derivatives and conjugates made therefrom

Also Published As

Publication number Publication date Type
CA1158557A (en) 1983-12-13 grant
EP0038357A1 (en) 1981-10-28 application
CA1158557A1 (en) grant
EP0038357A4 (en) 1982-03-22 application

Similar Documents

Publication Publication Date Title
Nakatsuka et al. Peptide segment synthesis catalyzed by the semisynthetic enzyme thiolsubtilisin
Groot et al. Anticancer prodrugs for application in monotherapy targeting hypoxia, tumor-associated enzymes, and receptors
US6703381B1 (en) Methods for delivery therapeutic compounds across the blood-brain barrier
US4963655A (en) Boron analogs of amino acid/peptide protease inhibitors
US5286637A (en) Biologically active drug polymer derivatives and method for preparing same
US4950738A (en) Amine derivatives of anthracycline antibiotics
US5866679A (en) Peptides
US6214345B1 (en) Lysosomal enzyme-cleavable antitumor drug conjugates
US4703107A (en) Water-soluble acylated derivatives of peptides or amino acids, their preparation and their use
US6265540B1 (en) Tissue specific prodrug
US6143716A (en) Liposomal peptide-lipid conjugates and delivery using same
US5554728A (en) Lipid conjugates of therapeutic peptides and protease inhibitors
US5502035A (en) N-terminus modified analogs of LHRH
US3998799A (en) Novel, transient pro-drug forms of l-dopa
US6759509B1 (en) Branched peptide linkers
US6034058A (en) Semi-synthetic alanyl dilemnin analogs
Lyttle et al. Isoenzyme-specific glutathione-S-transferase inhibitors: design and synthesis
US6034066A (en) Cysteine protease inhibitors for use in treatment of IGE mediated allergic diseases
US6472507B1 (en) Carrier based drug delivery system
US6613879B1 (en) FAP-activated anti-tumour compounds
US5624894A (en) Brain-enhanced delivery of neuroactive peptides by sequential metabolism
US5094848A (en) Cleavable diphosphate and amidated diphosphate linkers
US20020142955A1 (en) Enzyme-cleavable prodrug compounds
US4350627A (en) Biologically active peptides
US4318904A (en) Peptide affinity labels for thrombin and other trypsin-like proteases

Legal Events

Date Code Title Description
AK Designated states

Designated state(s): JP

AL Designated countries for regional patents

Designated state(s): CH DE FR GB NL

WWE Wipo information: entry into national phase

Ref document number: 1980902253

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1980902253

Country of ref document: EP

WWR Wipo information: refused in national office

Ref document number: 1980902253

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1980902253

Country of ref document: EP