MXPA98009760A - Therapeutic uses of bpi protein products in human beings with bleeding due to tra - Google Patents

Abstract

The present invention relates to the use of a therapeutically effective amount of a product of the bactericidal enhancing permeability protein (BPI), in the preparation of a medicament for treating hemorrhage due to a trauma in a human being.

Description

"THERAPEUTIC USES OF BPI PROTEIN PRODUCTS IN HUMAN BEINGS WITH HAEMORRHAGE DUE TO TRAUMA" This is a continuation application in part of U.S. Application Serial Number 08 / 652,292, filed May 23, 1996, which is incorporated herein by reference.

BACKGROUND OF THE INVENTION The present invention relates generally to methods and materials for treating humans suffering from haemorrhage due to trauma, by administration of protein products (BPI) that increase the bactericidal activity / permeability. Acute traumatic hemorrhage, which usually requires immediate surgical intervention, is a major contributor to morbidity and mortality in the United States [Bickell et al., New Eng. J. Med., 331: 1105-1109 (1994). Tran and others. Surgery, 114: 21-30 (1993).] In 1982, they had approximately 165,000 deaths in the United States due to trauma, with at least two additional cases of permanent disability for each death. Approximately 50 percent of these traumatic deaths occur immediately, due to direct injury to the central nervous system, heart or one of the major blood vessels. In addition, early deaths, approximately 30 percent, occur within several hours after the injury, usually due to uncontrolled bleeding. The remaining 20 percent of deaths are so-called "late deaths," which occur for days to weeks after the injury, due to complications of traumatic bleeding including infection or fainting of the multiple organ system (MOSF) in about 80 percent of the cases. [Trunkey, Sci. Am., 249: 28-35 (1983), Trunkey, New Eng. J. Med., 324: 1259-1263 (1991).] Among those patients who survive immediate resuscitation and surgical interventions, approximately 10 percent a 40 percent suffer from a variety of morbidities, including, for example, systematic inflation, wound infections, pneumonia, sepsis, respiratory failure, renal failure, coagulopathy and pancreatitis. Bleeding and transfusion requirements can be specifically linked to the increased risk of postoperative infection, respiratory complications, and multiple organ system failures [Agarwal et al., Arch, Surg. , 128: 111-111 (1993), Duke et al., Arch. Surg. , 128: 1125-1132 (1993), Tran et al., Supra].

The causes of these complications of traumatic hemorrhage are ultifactorial and interrelated. Many morbidities can be related to systematic inflammation after the injury. It has also been hypothesized that physical trauma to tissue, direct tissue hypoperfusion and translocation of endogenous bacteria and absorption of endotoxin from the bowel cavity (due to hypoperfusion and / or other damage to the gastrointestinal tract) may have a role in the fl 10 pathogenesis of these complications. The importance of these proposed factors in the pathophysiology of morbidities and late deaths associated with acute hemorrhagic shock in humans, however, is unclear. 15 Even though acute traumatic hemorrhage is a potential cause of hypovolemic shock (ie, shock due to decreased intravascular volume), there are numerous other potential causes, such as internal bleeding, e.g., gastrointestinal hemorrhage, hemorrhage intraperitoneal or retroperitoneal, hemorrhage towards the compartment of the femoral, intrathoracic hemorrhage, aortic dissection and ruptured aortic aneurysm; excessive fluid loss due to e.g., serious vomiting due to intestinal or pyloric obstruction, serious diarrhea, sweating, dehydration, excessive urination (due to diabetes mellitus, diabetes insipidus, excessive diuretics or the diuretic phase of acute renal failure), peritonitis, pancreatitis, planic ischemia, gangrene, burns; vasodilation due to e.g., damage to the central nervous system, anesthesia, ganglionic and adrenergic blockers, barbiturate overdose, poisons; and toxic or humoral metabolic vasodilation, such as acute adrenal insufficiency, or an anaphylactic reaction. Other causes of shock unrelated to the loss of circulatory volume include cardiogenic shock (e.g., acute myocardial infarction, tamponade or heart block) and obstructive shock (e.g., acute pulmonary embolism). [See, e.g., Manual of Medical Therapeutics, 28 Edition, Ewald et al., Editors, Little, Brown and Company, Boston (1995); Cecil's Textbook of Medicine, 17th Edition, Wyngaarden et al., Editors, W.B. Saunders Co. , Philadelphia (1985).] As will be noted below in Table I, presented, a normal individual can rapidly lose up to 20 percent of the blood volume without signs or symptoms. Limited signs of cardiovascular distress appear with losses of up to 30 percent of the blood volume, but the signs and symptoms of hypovolemic shock usually appear when the blood loss exceeds 30 percent to 40 percent of the volume of the blood. blood. Table I Percentage Quantity Clinical Manifestations of Lost Volume of Lost Blood (ml) -20% 500-1000 Usually no possible mild postural hypotension and tachycardia in response to exercise; vasovagal syncope can occur in 5% of cases -30% 1000-1500 Few supine changes occur; head lightness and hypotension commonly occur when standing; noticeable tachycardia in response to efforts 0-40% 1500-2000 Blood pressure, cardiac output, central venous pressure and urine volume are reduced even when supine position can be observed; thirst, shortness of breath, sticky skin, sweat, turbidity of consciousness and rapid rapid pulse 0-50 2000-2500 Serious shock, often resulting in death The patient is often oliguric, with urine yield of less than 20 milliliters per hour. Frequently, physical discoveries follow a progressive pattern as the shock leaves the early compensated phase to the advanced stages. In Stage I, physiological compensatory mechanisms, such as increased cardiac output or elevated systematic vascular resistance, symptoms and clinical signs are effective and minimal. In Stage II, these mechanisms can not effectively compensate for the loss of blood volume, and the patient may exhibit hypotension, tachycardia and hyperventilation. Decreased perfusion of vital organs can result in an altered mental state ranging from agitation to stupor to coma, reduced urine output, and iocardiac ischemia (in patients with coronary artery disease). The external appearance of the patients also reflects the excessive sympathetic discharge, with cyanosis, coldness and stickiness of the skin. In Stage III, which may be irreversible, excessive and prolonged reduction of tissue perfusion leads to significant alterations in cell membrane function, aggregation of red blood cells and "sediment" in the capillaries. The vasoconstruction that has taken place in less vital organs in order to maintain blood pressure is now excessive and has reduced flow to a degree where cell damage occurs. After traumatic hemorrhage, conventional therapy is aimed at stopping the hemorrhage, fighting the shock and restoring the blood volume. Resuscitation of the fast fluid is preferably provided through large perforation probes placed in the large peripheral veins. A pneumatic counter-pneumatic garment, with inflation in sequence of the legs, and abdominal compartments of up to 15 to 40 millimeters of mercury, can temporarily stabilize patients by increasing peripheral vascular systematic resistance. The restoration of blood volume can be achieved by intravenous infusion of electrolyte solutions; colloidal solutions of plasma protein albumin or dextran; or fresh whole blood. In an emergency situation, electrolyte, albumin or dextran solutions are preferred in relation to whole blood because large amounts of fluid are required, the possible delay in transfusion if typing and cross-matching are carried out and the possibility of allergic transfusion reactions. When the shock is due to hemorrhage, packed red blood cells should be provided as soon as possible. When the hemorrhage is massive, blood of a specific mismatched type can be safely provided. Rarely, type O blood may be needed. Rapid infusion of lactated or normal Ringer's saline solution is the most widely used fluid therapy after hemorrhage. An initial infusion of two to three times the volume of blood loss calculated is administered. Because these solutions are rapidly distributed through the intravascular and extravascular compartments, they must be supplemented with colloidal solutions. When large volumes of electrolyte solutions are provided, patients often develop peripheral edema and elderly patients may develop pulmonary edema. Colloidal preparations that are widely used include a 6 percent solution of high molecular weight dextran (dextran 70), a 10 percent solution of low molecular weight dextran (dextran 40), and a 5 percent solution of albumin in normal saline. Dextran 70 infusions produce an initial volume effect slightly greater than the amount that has been infused. Dextran 70 is cleared slowly over one or two days, allowing time for normal physiological mechanisms to replace the loss of volume. Dextran 40 has the advantage of an initial volume effect of almost double the amount infused. The lower molecular weight material quickly clears, however, and the volume expansion effect dissipates by 24 hours, before the normal volume replacement mechanisms reach their maximum. Acute renal failure has occurred in a few patients receiving dextran 40. With any dextran solution, volumes in excess of one liter can interfere with platelet adhesiveness and the normal coagulation cascade. A 5 percent solution of albumin in normal saline has the advantage of producing a known volume effect in hypovolemic patients, but this preparation is relatively expensive and delayed to prepare. A hypertonic albumin preparation containing 120 milliequivalents of sodium lactate, 120 milliequivalents of sodium chloride, and 12.5 grams of albumin per liter provides a predictable volume effect and minimizes leakage of interstitial fluid. The use of hypertonic solutions requires careful monitoring of central arterial and venous pressures to avoid fluid overload. Coexisting problems such as congestive heart failure, valvular heart disease, myocardial ischemia, or renal failure should be carefully monitored, and invasive hemodynamic monitoring should be considered during acute administration. The associated coagulopathy and the electrolyte imbalance can also be corrected. BPI is a protein isolated from granules of mammalian polymorphonuclear leukocytes (PMSs or neutrophils), which are essential blood cells in defense against invading microorganisms. The human BPI protein has been isolated from PMSs by extraction with acid combined with either ion exchange chromatography [Elsbach, J. Bi ol. Chem. , 254: 11000 (1979))] or affinity chromatography of E. coli [Weiss, et al., Bl ood, 69: 652 (1987)]. The BPI obtained in this manner is referred to herein as natural BPI and has been shown to have potent bactericidal activity against a broad spectrum of gram-negative bacteria. The molecular weight of human BPI is approximately 55,000 daltons (55 kD). The amino acid sequence of the entire human BPL protein and the DNA nucleic acid sequence encoding the protein have been disclosed in Figure 1 of the article by Gray et al., J. Biol. Chem. , 264: 9505 (1989), which is incorporated herein by reference. The amino acid sequence of Gray et al. Is set forth in SEQ ID NO: 1 appended hereto. U.S. Patent No. 5,198,541 discloses recombinant genes that encode methods for the expression of BPI proteins, including holoprotein BPI and BPI fragments.

BPI is an intensely cationic protein. The N-terminal half of BPI is responsible for the high net positive charge; the C-terminal half of the molecule has a net charge of -3. [Elsbach and Weis (1981), supra]. The N-terminal proteolytic BPI fragment having a molecular weight of about 25 kD possesses essentially all of the anti-bacterial efficacy of the naturally derived 55 kD human BPI holoprotein. [Ooi et al., J. Bio. Chem., 262: 14891-14894 (1987)]. In contrast to the N-terminal portion, the C-terminal region of the isolated human BPI protein exhibits only slightly detectable antibacterial activity against gra-negative organisms. [Ooi et al., J. Exp. Med., 1 74: 649 (1991).] The approximately 23 kD N-terminal BPI fragment referred to as "rBPl23" has been produced by recombinant means and also retains the antibacterial activity against gram-negative organisms. Gazzano-Santoro and others, Infect. Immun. 60: 4754-4761 (1992). The bactericidal effect of BPI has been reported to be highly specific for the gram-negative species, eg, in Elsbach and Weiss, Inflammation: Basic Principles and Clinical Correlates, editors, Gallin et al., Chapter 30, Raven Press, Ltd. (1992). The precise mechanism by which the BIS kills the gram-negative bacteria has not yet been fully elucidated but it is believed that the BPI must first be ligated to the surface of the bacterium through electrostatic and hydrophobic interactions between the cationic BPI protein and the sites negatively charged in LPS. In susceptible gram-negative bacteria, BPI binding is thought to break the structure of LPS, leading to the activation of bacterial enzymes that degrade phospholipids and peptidoglycans, altering the permeability of the cell's outer membrane, and initiating events that ultimately lead the death of the cell. [Elsbach and Weiss (1992), supra]. LPS has been termed as an "endotoxin" due to the potent inflammatory response that it stimulates, ie, the release of mediators through host inflammatory cells that can ultimately result in irreversible endotoxic shock. BPI binds to lipid A, which has become known as being the most toxic and most biologically active component of LPS. The BPI protein has never been used before for the treatment of humans suffering from haemorrhage due to trauma or shock associated with traumatic blood loss (i.e., hypovolemic shock). Bahrami et al., Presentation at the Board of the International Endotoxin Society of Vienna, August, 1992, disclose the administration of the BPI protein to rats subjected to hemorrhage. Yao and others, Ann. Surg. , 221: 398-405 (1995), disclose the administration of rBPl2i (described infra) to rats subjected to prolonged hemorrhagic insult for 180 minutes followed by resuscitation. U.S. Patent Nos. 5,171,739, 5,089,724 and 5,234,912 disclose the use of BPI in various studies of animal models in vi tro and in vivo that are confirmed to be related to methods to treat endotoxin-related diseases, including endotoxin-related shock. In co-owned joint US applications Number 08 / 378,228, filed on January 24, 1995, Number 08 / 291,112, filed on August 16, 1994, and Number 08 / 188,221, filed on January 24, 1994, which are incorporated herein by reference, describe the administration of the BPI protein product to humans with endotoxin in the circulation. [See von der Mohlen et al., J. Infect. Dis. 1 72: 144-151 (1995); von der Mohlen et al., Blood 85: 3437-3443 (1995); by Sinter and ~ others, J. Inflam. 45: 193-206 (1995)]. In the co-pending joint US application Serial Number 08 / 644,287, filed May 10, 1995, the administration of the BPI protein product to humans suffering from serious meningococcemia is described.

Despite treatment with antibiotics and medical intensive care therapy according to the current state of the art, the mortality and morbidities associated with bleeding due to trauma remain significant and have not been resolved by current therapies. New therapeutic methods are needed that can reduce or alleviate the harmful events and improve the clinical performance of these patients.

COMPENDIUM OF THE INVENTION The present invention provides novel methods for treating humans suffering from trauma-related hemorrhage, involving the administration of BPI protein products to provide clinically verifiable relief of the detrimental effects or, of complications associated with this disease state, including mortality and complications or morbidities. In accordance with the invention, BPI protein products such as rBPl2i are administered to humans suffering from acute traumatic hemorrhage in sufficient quantities to reduce or prevent mortality and / or to reduce the incidence (i.e., the event) or seriousness of complications or morbidities, including infection (eg, infection of the surgical site) or organ malfunction (eg, disseminated intravascular coagulation, acute respiratory distress syndrome, acute renal failure or hepatic malfunction). The use of a BPI protein product in the preparation of a medicament for the treatment of humans suffering from haemorrhage due to trauma is also proposed. The numerous additional aspects and advantages of the invention will become apparent to those skilled in the art upon taking into account the following detailed description of the invention which describes the presently preferred embodiments thereof.

BRIEF DESCRIPTION OF THE FIGURES Figure 1 shows the incidence of harmful events in rBPl2i and placebo treatment groups.

DETAILED DESCRIPTION Acute hemorrhage due to trauma is a life-threatening condition with significant morbidity and mortality despite intensive medical care according to the current state of the art. The administration of BPI protein products to humans suffering from haemorrhage due to acute traumatic injury (such as penetration and / or trauma) is to be expected to effectively decrease mortality and reduce the incidence (i.e., the event) or seriousness of complications or morbidities associated with or resulting from hemorrhage due to trauma. Complications include infection (eg in surgical sites, wounds, organs, anatomical spaces, bloodstream, urine conduit or pneumonia) or organ malfunction (eg, disseminated intravascular coagulation, acute respiratory distress syndrome (ARDS), acute renal failure). or hepatobiliary malfunction), and may include serious complications. An additional complication may be the pulmonary malfunction that includes ARDS and pneumonia. These unexpected effects on mortality and complications associated with and resulting from trauma hemorrhage indicate that BPI protein products effectively interfere with or block a number of poorly understood multiple pathophysiological processes that have led to poor performance in this condition . BPI protein products can be used as adjunctive therapy in the treatment or prevention of organ malfunction and serious infections. The BPI protein products are expected to provide beneficial effects for patients suffering from haemorrhage due to trauma, such as reduced injury seriousness score, reduced time interval in ventilatory and inotropic support therapy (vasoactive), reduced duration or severity of associated coagulopathy, reduced ICU stay, reduced hospital stay, reduced incidence and duration of complications such as coagulopathy, respiratory faintness, renal failure, hepatic failure, or altered mental status, adrenal cortical necrosis, and serious infection, including in wounds, organs, anatomical spaces and blood streams, the urine duct or pneumonia. The therapeutic compositions comprising the BPI protein product can be administered systematically or topically. Systematic routes of administration include oral, intravenous, intramuscular or subcutaneous injection (including to a depot for long-term release), infra-ocular and retrobulbar, intrathecal, intraperitoneal (eg, by intraperitoneal lavage), intrapulmonary injection using an aerosolized or nebulized drug or - l Transdermal The preferred route is intravenous administration. When provided parenterally, compositions of the BPI protein product are usually injected at doses ranging from 1 microgram per kilogram to 100 milligrams per kilogram per day, preferably doses ranging from 0.1 milligram per kilogram to 20 milligrams per kilogram per kilogram per kilogram per kilogram per day. day, more preferably doses ranging from 1 to 20 milligrams per kilogram per day and especially preferably at doses ranging from 2 to 10 milligrams per kilogram per day. Treatment may be initiated immediately after the trauma or within the period of time subsequent to the trauma (including, eg, within 6, 12 or 24 hours after the trauma, or within a reasonably clinical time period that is determined by the doctor who carries out the treatment, for example 48 to 72 hours after the trauma). A continuous intravenous infusion of the BPI protein product at a dose of 4 to 6 milligrams per kilogram per day is currently preferred, continuing for 48 to 72 hours. The treatment may be continued by continuous infusion or intermittent injection or infusion at the same dose or a reduced or increased dose per day, for example e.g., from 1 to 3 days, and further as determined by the treating physician. Alternatively, the BPI protein products are administered intravenously by a large initial ball followed by continuous infusion. One of these regimens is a large intravenous pill of 1 to 20 milligrams per kilogram of the BPI protein product followed by intravenous infusion at a dose of 1 to 20 milligrams per kilogram per day, continuing for up to one week. Another dosage regimen is an initial large pill of 2 to 10 kilograms per kilogram followed by intravenous infusion at a dose of 2 to 10 milligrams per kilogram per day, continuing for up to 72 hours. Topical routes include administration in irrigation fluids for, v. g., Irrigation of wounds, or intrathoracic or intraperitoneal cavities. Other topical routes include administration in the form of ointments or ointments, eye drops, ear drops or medicated shampoos. For example, for topical administration in the form of drops, about 10 to 200 microliters of the composition of the BPI protein product may be applied one or more times per day, as determined by the treating physician. Those of ordinary skill in the art can easily bring effective dosing and administration regimens to therapeutic compositions comprising the BPI protein product, as determined by good medical practice and the clinical condition of the individual patient.

As used herein, the term "BPI protein product" includes the BPI protein produced naturally and recombinantly; and recombinant biologically active polypeptide fragments of the BPI protein, biologically active polypeptide variants of the BPI protein or fragments thereof including proteins and hybrid fusion dimers; biologically active polypeptide analogues of BPI protein or fragments or variants thereof, including analogs substituted with cysteine; and peptides derived from BPI. The BPI protein products administered in accordance with this invention can be generated and / or isolated by any means known in the art. U.S. Patent No. 5,198,541, the disclosure of which is incorporated herein by reference, discloses recombinant genes encoding and methods for the expression of BPI proteins including recombinant BPI holoprotein, referred to as rBPl5Q (or rBPI) and recombinant BPI fragments. The co-pending US patent application Serial Number 07 / 885,501 and a continuation in part thereof, US Patent Application Serial No. 08 / 072,063 filed May 19, 1993, and the corresponding PCT application No. 93/04752 filed May 19, 1993, all of which are hereby incorporated by reference, disclose novel methods for the purification of recombinant BPI protein products expressed in and secreted from genetically transformed mammalian host cells in culture and discloses the manner in which large quantities of suitable recombinant BPI products can be produced to be incorporated into homogeneous pharmaceutical preparations stable Biologically active BPI fragments (BPI fragments) include biologically active molecules that have an amino acid sequence that is the same or similar to a natural human BPI haloprotein, with the exception that the fragment molecule lacks amino-terminal amino acids, internal amino acids, and / or carboxy-terminal amino acids of holoprotein. Non-limiting examples of these fragments include a N-terminal fragment of native human BPI of approximately 25 kD, which is described in the article by Ooi et al., J. Exp. Med., 1 74: 649 (1991), and recombinant expression product of N-terminal amino acids encoding DNA from 1 to approximately 193 or 199 of natural human BPI, which is described in the article by Gazzano-Santoro et al., Infect. Immun. 60: 4754-4761 (1992), and to which reference is made as (rBPl23) - In this publication, an expression vector was used as a source of DNA encoding a recombinant expression product (rBPl23) having the sequence of 31-residue signal and the first 199 amino acids of the N-terminus of mature human BPI, as noted in Figure 1 of the article by Gray et al., supra, with the exception that valine at position 151 is specified by GTG instead of GTC and residue 185 is glutamic acid (specified by GAG) instead of lysine (specified by AAG). The recombinant holoprotein (rBPl5n) has also been produced having the sequence (SEQ ID NOS: 1 and 2) which is pointed out in Figure 1 of the article by Gray et al., Supra, with the exceptions mentioned for rBPl23 and with the exception that residue 417 is alanine (specified by GCT) instead of valine (specified by GTT). Other examples include the dimeric forms of the BPI fragments, as described in co-pending US Patent Application and co-owned Serial Number 08 / 212,132, filed March 11, 1994, and the corresponding PCT Application PCT Number. / US95 / 03125, the exhibits of which are incorporated herein by reference. Preferred dimeric products include the dimeric BPI protein products wherein the monomers are amino-terminal BPI fragments having the N-terminal residues from about 1 to 175 to about 1 to 199 of the BPI holoprotein. A particularly preferred dimeric product is the dimeric form of the BPI fragment having N-terminal residues from 1 to 193 designated the dimer of rBPl42 >; Biologically active variants of BPIs (BPI variants) include but are not limited to recombinant hybrid fusion proteins comprising the BPI holoprotein or the biologically active fragment thereof and at least a portion of at least one other polypeptide; Dimeric forms of the BPI variants. Examples of these hybrid fusion proteins and dimeric forms are described by Theofan and others in co-pending US Patent Application Serial No. 07 / 885,911, and a co-owned application thereof, the Patent Application. US Serial No. 08 / 064,693 filed May 19, 1993 and the corresponding PCT Application Number US93 / 04754 filed May 19, 1993, all of which are incorporated herein by reference include hybrid fusion proteins that comprise the amino-terminal end, a BPI protein or a biologically active fragment thereof, and the carboxy terminal end of at least one constant domain of an immunoglobulin heavy chain or an allelic variant thereof. Hybrid fusion proteins similarly configured involve part or all of the Lipopolysaccharide Ligament Protein (LBP) and are also proposed for use in the present invention. Biologically active analogs of BPI (BPI analogs) include but are not limited to BPI protein products wherein one or more of the amino acid residues have been replaced by a different amino acid.

For example, the co-pending Co-pending US Patent Application Serial Number 08 / 013,801 filed on February 2, 1993 and the corresponding PCT Application Number US94 / 01235 filed on 2 February 1994, the teachings of which are incorporated herein by reference, disclose BPI polypeptide analogs and BPI fragments wherein a cysteine residue is replaced by a different amino acid. A preferred BPI protein product described by this application is the DNA expression product encoding amino acid 1 to about 193 or 199 of the N-terminal amino acids of the BPI holoprotein, but where the cysteine in the residue number 132 is replaced with alanine and is designated rBPl2i? Cys or rBPl2 ?.

Other examples include the dimeric forms of the BPI analogs; e.g., the patent application North American co-owned and co-owned Number of Series 08 / 212,132, filed on March 11, 1994, and the corresponding PCT application Number PCT / US95 / 03125, the exhibits of which are incorporated herein by reference. Other BPI protein products useful in accordance with the methods of the invention are peptides derived from or based on BPI produced by recombinant or synthetic means (BPI-derived peptides), such as those described in the copending and co-pending US Patent Application. -property Serial Number 08 / 504,841, filed July 20, 1995 and in the co-pending and co-owned PCT Application PCT / US94 / 10427 filed September 15, 1994, which corresponds to the North American Patent Application Serial Number 08 / 306,473 filed September 15, 1994, and PCT Application Number US94 / 02465 filed March 11, 1994, corresponding to US Patent Application Serial No. 08 / 209,762 filed March 11 of 1994, which is a continuation in part of the US Patent Application Serial Number 08 / 183,222, filed on January 14, 1994, which is a continuation in part of U.S. Patent Application Serial No. 08 / 093,202, filed July 15, 1993 (and for which the corresponding international application is PCT Application Number US94 / 02401 filed March 11, 1994), which is a In the following part of the US Patent Application Serial Number 08 / 030,644 filed on March 12, 1993, the exhibits of which are incorporated herein by reference. Currently preferred BPI protein products include the recombinantly produced N-terminal fragments of BPI, especially those having a molecular weight of approximately 21 to 25 kD such as rBPl23 or rBPl2i, or the dimeric forms of these N-terminal fragments (eg dimer rBPl42). In addition, preferred rBPI protein products include rBPl5Q and peptides derived from BPI. RBPI2 ?. Administration of the BPI protein products is preferably achieved with a pharmaceutical composition comprising a BPI protein product and a pharmaceutically acceptable diluent, an adjuvant or a carrier. The BPI protein product can be administered without or in conjunction with known surfactants, other chemotherapeutic agents or additional known anti-microbial agents. A pharmaceutical composition containing BPI protein products (v.gr, rBPl5Q, rBPl23) comprises the BPI protein product at a concentration of 1 milligram per milliliter in citrate stabilized saline (5 or 20 mM citrate, 150 M NaCl , pH 5.0) comprising 0.1 weight percent poloxamer 188 (Pluronic F-68), BASF Wyandotte, Parsippany, NJ) and 0.002 weight percent polysorbate 80 (Tween 80, ICI Americas Inc., of Wilmington, DE). Another pharmaceutical composition containing BPI protein products (e.g., rBPl2i) comprises the BPI protein product at a concentration of 2 milligrams per milliliter in 5 M citrate, 150 mM NaCl, 0.2 percent poloxamer 188 and 0.002 percent polysorbate 80. These combinations are described in the co-pending PCT Application co-owned US94 / 01239 filed February 2, 1994, which corresponds to US Patent Application Serial No. 08 / 190,869 filed on 2 February 1994, and U.S. Patent Application Serial No. 08 / 012,360 filed February 2, 1993, the exhibits of which are incorporated herein by reference. Other aspects and advantages of the present invention will be understood by taking into account the following illustrative examples. Example 1 is directed to the effect of the administration of the BPI protein product in humans on mortality and complications associated with bleeding due to trauma.

EXAMPLE 1 Protocol of the Clinical Study - Therapeutic Effects of the BPI Protein Product A human clinical study was designed to examine the effect of an exemplary BPI protein product, rBPl2 ?, in the treatment of patients with acute hemorrhage due to trauma. Therefore, a multicenter, randomized, double-blind placebo-controlled trial was implemented comparing placebo treatment and rBP12 treatment? provided to more than 48 hours in patients with acute hemorrhage due to trauma. Approximately 400 patients admitted to the emergency department with acute bleeding due to trauma and requiring transfusion of at least two units of blood were randomly placed in a 1: 1 ratio for treatment with either rBPl2? or placebo. In addition to normal therapy, each patient received continuous intravenous infusion, either rBPl2? to 8 milligrams per kilogram through 48 hours (4 milligrams per kilogram per day for 2 days) or an equivalent volume of placebo. In most cases, the patient's weight in kilograms was determined as a good calculation. Efficacy was monitored from Day 1 to Day 15 following patients for development of complications, such as function and infection of damaged organs and for survival. Safety was monitored through pretreatment and post-treatment tests in series of chemical parameters and hematology, as well as daily assessments for damaging events until Day 15. A final assessment of survival and harmful complications occurred on Day 29. Patients taken to the hospital with acute hemorrhage due to trauma were selected to be admitted to the study, if they met the following inclusion and exclusion criteria. The inclusion criteria were: (X) age 18 years (or age of consent) up to 75 years, inclusive. (2) patient suffering from acute hemorrhage secondary to trauma; (3) study drug provided within 6 or 12 hours of the occurrence of the traumatic event (if the precise time of the event was unknown, the best calculation was provided); (4) the patient requires and has begun to receive a second unit of packed red blood cells; and (5) the patient provides verbally informed consent or a relative provides written informed consent. The exclusion criteria were: (1) a Revised Selection Trauma Score (TRTS, scale of 0 to 12) less than 2.0 during admission to the Emergency Department, see Table II below [Champion et al., Cri t. Care Med., 9 (9): 672-676 (1981); Greenfiel et al., Chapter 10, in Surgery Scientific Principies and Practices, J.V. Lippincortt Co., Philadelphia, pages 252 to 255 (1993)]; (2) serious head trauma (Glasgow Coma Score (GCS) <5 or equivalent evidence), see Table III below [Teasdale et al., Lancet, 1: 81 (1974)]; (3) isolated cranial injury; (4) spinal injury with paralysis; (5) burn injuries with at least 20 percent of the body surface area with second degree burn; (6) Known positive HIV (test not prescribed during Xa entry); (7) known pre-existing kidney disease (creatinine> 2.0); (8) Known pre-existing heart disease (NY Heart Association class greater than III, see Table IV below) [Braunwald, in Braunwald et al., Heart Disease, The Texbook of Cardi ovascular Medicine, Third Edition, W.B. Saunders Company, Philadelphia, PA, page 12 (1988); J. Am. Med. Ass' n, 249: 539-544 (1988)]; (9) pre-existing primary or metastatic malignancy known in visceral organs; (10) arterial pH (at initial evaluation) < 6.8 or base deficit > 15 (if measured); (11) known current spheroid therapy (> 10 milligrams of prednisone per day for > one month); (12) active hepatitis or known pre-existing cirrhosis; (3) pregnancy or lactation; (14) participation in other research drug studies (including research blood products) within the previous 30 days; (15) weight (calculated) greater than 120 kilograms; and (16) a "do not resuscitate" (DNR) or equivalent order. Table II Revised, Selected Trauma Score (TRTS) * VALUATION METHOD CODIFICATION System regimen breathing rate10-29 4 Thorium Respiratory in 15 seconds and > 29 3 (RR) multiply by 4 6-9 2 1-5 1 0 0 Pressure The systolic pressure is measured > 89 4 Blood either in the arm through 76-89 3 Systolic (SBP) auscultation or palpitation 50-75 2 1-49 1 0 0 Score is calculated according to Covierta GCS at Glasgow Coma Table III below: (GCS) presents below 13-15 = 4 9-12 = 3 6-8 = 2 4-5 = 1 < 4 = 0 * The TRTS is the sum of the codes for RR, SBP and GCS (scale from 0 to 12).

Table III Glasgow Coma Scale * Spontaneous Eyes Opening 4 Sound Response 3 Pain Response 2 Never 1 Response Motor Obecer controls 6 Localized pain 5 Normal flex 4 (withdrawal) Abnormal flex 3 (decorticate) No response 1 Verbal Response Oriented 5 Confused conversation 4 Inappropriate words 3 Incomprehensible sounds 2 None 1 * Scores vary from 3 to 15 Table IV Modified Functional Classification of the New York Heart Association Class I. Patients with heart disease but without any limitation of physical activity. Ordinary physical activity does not cause undue dyspnea, angina pain, fatigue, or palpitation. Class IIS. Patients with mild limitation of physical activity. They were comfortable during the break and with moderate effort. They experience symptoms only with the heavier regular activity classes. IIM class. Patients with moderate physical capacity limitation. They were comfortable during the break and with gentle effort. They experience symptoms with moderate degrees of regular activity. Class III Patients with noticeable limitation of physical activity. They are comfortable during rest but experience symptoms with even milder forms of regular activity. Class IV. Patients with the ability to carry out any physical activity without discomfort. Symptoms of heart failure or angina syndrome may be present, even during rest and intensified by activity - The following were recorded for all patients placed at random for treatment: (1) date and estimated time of the incident, and date and time of admission to the Emergency Department; (2) for patients placed at random and not treated, the reason for not being treated; (3) from arrival at the hospital until approximately 48 hours post-operatively, date, time, volume and location in which the patient received blood, blood products and fluids such as packed red blood cells, whole blood, self-transfusion, platelets, fresh frozen, crystalloid, or colloid plasma, in locations such as the Emergency Department, the Operating Room, the Post-Anesthesia Care Unit, or the Quirugica Intensive Care Unit; however, if the patient did not undergo surgery, the aforementioned times were applicable and were collected during study days 1, 2 and 3; (4) date and time in which the second unit of blood was administered (which should have preceded the surgery, to ensure that the bleeding is due to trauma, not to surgery), and date and times of initiation and anesthesia arrest; (5) date and times of initiation and detention of surgery, blood loss calculated in the operating room, and date and time in the post-anesthesia care unit; (6) date and time when the infusion of the study drug began and ended, volume infused, and reasons for temporary or permanent discontinuation; if applicable, and if the amount that was provided was discontinued; (7) directed medical history (including degree and nature of injuries, interacting diseases, conditions that contribute to bleeding, etc.), demographic and physical examination information such as sex, age, weight (calculated or measured), height (calculated or measurement), vital signs, physical signs of injury; (8) results of the pregnancy test carried out during the selection for eligibility of appropriate female patients (all women with potential to have children, that is, all women who were not surgically sterile or documented to be cataloged as post-menstruation) and (9) the results of TRTS carried out during the selection for eligibility (including current measures). After the transfusion of the second unit of blood was initiated, the investigator administered an unknown test drug of kits in sequential numbered order. Each box already contained rBPl2? or placebo. The rBPl2? was supplied as a sterile non-pyrogenic colorless crystalline solution in X0 milliliters of small single use glass vials at a concentration of 2 milligrams per milliliter in 5 mM sodium citrate / 0.15 M sodium chloride stabilizer, pH of 5.0 and 0.2 percent poloxamer 188 and 0.002 percent polysorbate 80, which does not contain a preservative. The rBPl2? it was stored refrigerated at 2 ° to 8 ° C at all times before administration. Placebo was delivered as a non-pyrogenic sterile colorless crystal solution in small single-use glass bottles of 10 milliliter capacity. It was composed of 0.2 milligram per milliliter of human serum albumin in 5 mM sodium citrate / 0.15 M sodium chloride stabilizer, pH 5.0, which did not contain any preservative. The placebo was also stored refrigerated at a temperature of 2 ° to 8 ° C at all times before administration. The kit assigned to each patient contained a sufficient number of small bottles of study medication for all X doses for that patient. Each small vial contained 10 milliliters of the test article. The study was administered to two groups (rBPl2"active" and placebo control) as noted above. The study drug was brought to room temperature before the infusion. Through the dosing procedure, good aseptic technique was followed for intravenous administration. The study drug was administered by intravenous infusion into a peripheral central vein for more than 48 hours. The infusion bag / tubing administration set changed completely after 24 hours. Appropriability of intravenous access was determined by easy removal of blood from the access, as well as easy infusion of intravenous fluids without infiltration. The medication in the study was the only agent administered in the orifice selected during the course of the infusion protocol. The venous access orifice was not heparinized, but was washed as needed without physiological saline. Any sign of a reaction at an infusion site was recorded in the form of a patient case record and a source document as a damaging experience. Patients treated at the selected study sites were assessed for: (1) blood levels of rBPl2 ?: blood for the evaluation of rBPl2 level? which is removed during Xos following periods of time (at selected study sites only): before the start of the infusion (up to 60 minutes before the start of the infusion), the following times (hours) after the start of the infusion; 1, 4, 8, 12, 20, 24, 32, 36, 40, within 15 minutes before completion of infusion of 48 hours, and the following periods of time after the infusion is completed; 7 minutes (48:07), 15 minutes (48:15), 30 minutes (48:30), X hour (49.00), 3 hours (51:00), 6 hours (54:00), and 24 hours ( 72:00); (2) antibodies to rBPl2 ?: the blood for evaluation of antibodies to rBPl2? it is removed at selected study sites during the following time periods: Day X before the study of the drug infusion, and Days 15 and 29, if the patient is still in the hospital or returns to the clinic (the days of extraction current may vary from Days 10-20 and Days 2X-29); and (3) cytokines: the blood for cytokine evaluation is extracted at selected study sites. The following safety laboratory panels were evaluated on Day 1 before the infusion of the test drug, Day 3 (after the end of the infusion) and Day 8, however, if the patient is discharged on or before on Day 8, the evaluation was made before the discharge if possible: (1) hematology jury: hemoglobin count, hematocrit, erythrocyte, leukocyte and differential count, and platelet count; (2) jury of serum chemistry: sodium, potassium, chloride, calcium, phosphorus, blood urea nitrogen, creatinine, uric acid, glucose (fasting), CPK, cholesterol, albumin, total protein, AST, ( SGOT), ALT (SGPT), bilirubin (total), GGT, LDH, and alacalin phosphatase. The following were recorded for all patients treated until Day 15 and / or Day 29 post-initiation of the drug infusion study: (1) adverse events continued until day 29); (2) current state of survival including the date and cause (s) of death (continued through Day 29); (3) dates in ICU (continued through Day 29); (4) dates in the hospital (continued through Day 29); (5) fan dates (continued through Day 29); (6) dates on dialysis or have an appointment, method of specification, (continued through Day 29); (7) concomitant medications, including daily amounts of transfused blood (continued until Day 15 or Day 29); (8) primary surgical procedures carried out for example including re-operations but excluding procedures such as central line placement, Swan-Ganz probes, arterial lines; lumbar punctures, etc., (continued until Day 15); (9) Seriousness of injury score (ISS) based on the diagnostic evaluations carried out during your stay in the current hospital; (10) daily evaluation of abnormal functions of the organ and the presence of infections (continued until Day 15); (11) daily vital signs associated with and including maximum daily temperatures and minimum daily temperatures (continued through Day 15); and (12) inspection of the infusion site used for administration of the study drug at least every eight hours, with observations documented in advance notes or the equivalent. Abnormal organ functions were evaluated using the following definitions. The patient is considered as having disseminated intravascular coagulation (DIC) when there were: (1) abnormally low values for platelets (or there was a decrease of> 25 percent of the value previously documented) and either a high prothrombin time or a thromboplastin time fl) partial elevated and clinical evidence of bleeding, or (2) if obtained, a confirmation test that was positive (FDP> 1:40 or D-Dímeros> 2.0). These abnormalities may have occurred in the absence of medically significant confounding factors such as faintness of the liver, primary hematoma, or anticoagulant therapy. The patient was considered as having an acute respiratory distress syndrome (ARDS) when: he had bilateral pulmonary infiltrates with pulmonary edema and Pa? 2 / Fi? 2 < 200. These signals must have occurred in the absence of faint congestive heart or primary lung disease such as pulmonary embolus or Pneumonia. The Pulmonary Artery Wedge Pressure (PAWP), when measured, must have been from < 18 millimeters of mercury.

The patient was considered as having acute renal failure (ARF) when: (1) dialysis or ophiltration was required (definition used for primary analysis) or (2) serum creatinine became abnormal with an increase of > 2.0 milligrams per dL in a patient with documented normal baseline creatinine or (3) serum creatinine was > 3.0 milligrams per dL in a patient who was not known to have renal insufficiency, but whose baseline creatinine (pretrauma) was unknown, or (4) serum creatinine doubled from admission or pre-treatment level. rBPl2? in a patient with anterior renal failure. These discoveries must not have been pre-renal in nature (e.g., associated with dehydration or gastrointestinal bleeding) or due to rhabdomyolysis. Abnormal post-surgical hepatobiliary function (HBD) was evaluated only in patients without primary liver disease (e.g., hepatitis or cirrhosis), alcoholism, or biliary disease. The patient was considered to have abnormal hepatobiliary function when: bilirubin exceeded 3.0 milligrams per dL, and either alkaline phosphatase, gamma-glutamyl transpetidase (GGT), alanine aminotransferase (ALT or SGPT) or aspartate aminotransferase (AST or SGOT) exceeded twice the upper limit of normal. These discoveries must have occurred in the absence of a detestable disease. Patients who were evaluated for wound infections, surgical sites (both superficial and deep incision sites), organs, anatomical spaces, the bloodstream (bacteremia), the urine duct, or the respiratory tract (pneumonia). Principal investigators of the physician were provided with the definitions of each abnormal function of the organ and were instructed to perform the registration during the pre-treatment and daily during Days 1-15 (a) and each abnormal function of the organ was "Present", " Clinically present "," Not present or clinically present ", or" Was unknown "according to the definitions that were provided and (b) all current available laboratory or clinical data required by the definition, whether or not it had been satisfied the definition. Researchers were also provided with definitions of infections and were instructed to register (a) and each type of infection was "Present" or "Not Present" at any time during Days 1-15 and (b) if "Present" the actual crop or clinical data required by the definition. In order to provide a more objective analysis of these points, and during the recommendation of an independent Data Security Supervision point, computer programs were developed before the first interim analysis of 50 percent cumulative efficacy to implement the same function Abnormal organ and definitions of infection using current laboratory data, clinical and culture required for each definition. The algorithmic approach defined each abnormal function of the organ as "Present", "Clinically present", "Not present or clinically present" or "Unknown" during pretreatment and daily during days 1-15. Each abnormal function of the organ was classified as "Unknown" on any given day if certain minimum evaluations required by the definition were not provided on that day. The definition for "Present" on a given day required that all evaluations have been made and that each evaluation should meet its respective criteria for the abnormal function of the specified organ. Therefore, patients for whom one or more required evaluations were missing on a given day could not be classified as having filled the definition for "Present" for that abnormal organ function for that day. On days when the evaluations were incomplete, the abnormal function of the organ was classified as "Clinically present" if each of the evaluations that did not fail filled their respective criteria for "Present" for that abnormal function of the organ. Therefore, "Clinically present" implies that the abnormal function of the organ may have been present that day, based on incomplete evidence, and that no contradictory evidence was recorded on that day. Since ARF would be defined as the use of dialysis / hemofiltration at least one day during Days 1-15, the term "Clinically present" was not applicable for serious ARF. An abnormal function of the organ was considered "Not Present" or "Clinically Present" when none of the definitions for "Present" "Clinically present" or "Unknown" were filled for that day. Patients who were classified as having an abnormal function of the "Present" or "Clinically present" pretreatment organ were ruled to have filled the primary point by another complication in order to have classified as having filled the primary point during Days 1-15 . For infections, the algorithmic approach defined each infection "Present" or "Not present" during Days 1-15. Each definition for "Present" r &ampHe requested that the definition be strictly filled in at least one day during Days 1-15 according to the data provided. If the patient did not fill the definition of infection, "Present", it was classified as "Not present" for that infection. Serious complications were defined as the event of the following serious infections: (1) deep incision surgical site infection, (2) anatomical or organ space infection, (3) secondary bloodstream infection, (4) an infection of the primary bloodstream; and (5) pneumonia; or the following serious abnormal organ functions: (X) disseminated intravascular coagulation (DIC) or coagulopathy, (2) acute respiratory distress syndrome (ARDS), (3) acute renal failure (ARF) requiring dialysis or hemofiltration, and (4) abnormal hepatobiliary function (HBD).

Patients were counted once as having complications, regardless of the number of complications. Across 19 sites, 14X1 patients were selected and 401 patients were placed randomly in the study groups and received therapy. Among these 401 patients, 199 received placebo treatment and 202 received rBPl2 treatment. Thirty-one patients (15 placebo, 16 rBPl2i) did not receive the full administration of the study drug. Twelve patients (six of placebo and six of rBPl2?) Were withdrawn from the study before Day 29. Data from all patients who received any amount of study medication, even when the infusion was incomplete, were included in all analyzes of safety and efficacy, whether or not the patient had been removed from the study. The average age of the study population was 35 (scale: 16-80 years); 80 percent of the patients were under 45 years old. The average dosage weight was 79 kilograms (scale: 45-145 kilograms). Seventy-seven percent of the patients were male. The source of traumatic injury was classified as blunt trauma (50 percent), penetrating trauma (48 percent), or both (2 percent). Of the other trauma-related characteristics, mean TRT was 10.6 (scale: 0-12); GCS medium was 13.2 (scale: 3-15); Mean ISS ('90 version) was 23.9 (scale: 1-75), the mean number of PRBC units that were started before the infusion of the study drug was 6.4 (scale: 0-57), and the time mean for the traumatic incident of the drug infusion was 9.5 hours (scale: 1.3-2X.8 hours). There were no significant treatment group differences for age, weight, ethnicity, source of injury, TRTS, GCS, and time for infusion (p> 0.X0 control for the site), but the placebo group had a somewhat higher proportion of female patients (P = 0.11, control for the site). The number of packed red blood cell (PRBC) units of transfusion before the infusion of the drug was • similar among patients placed at random for rBPl2? and placebo. The mean ISS was slightly worse (P = 0.07 5 control for the site) for the placebo patients (mean 25.1) than for the rBPl2 patients? (medium 22.7). The prespecified primary efficacy analysis focused on the primary end point of mortality or serious complication (defined as abnormal organ function serious or serious infection as assessed by the algorithmic approach) that occurred at any time after zero hour until Day 15. Total mortality in this study was low (approximately 5 percent to 6 percent). The treatment groups were compared using Cox regression, stratifying by site and not adjusted for covariates. The results were analyzed by computer algorithm using strict definitions of • "Present" and also incorporating incomplete evidence (leading to the classifications of "Present or Clinically Present ".) Regardless of the algorithmic method, the mortality regimen or serious complication by Day 15 was less than 7 percent for patients with rBP122 compared to placebo patients Kaplan-Meier calculations of regimens of event for mortality or serious complication using both algorithmic methods are shown below Table V. Table V Results and Statistical Calculations Placebo rBPI2? (N = 199) (N = 202) p value "Present" Percentage of Current Event Definition Regime by (# of patients with Algorithm events / # of events in 46% 39% total) (91/199) (78/202) Kaplan-Meier calculations of 15-day Event Regimes 46% 39% 0.17"Present / Percentage of Clinical Regime Current Present Event" (# of patients with Definition of events / by # of patients in 55% 48% Total Algorithm) (109/199) (97/702) Kaplan-Meier calculations of 15-day Event regimens 55% 48% 0.15 Placebo patients were approximately 1.27 times more likely to experience mortality or serious complications with rBPl2? by both algorithmic methods as measured by the risk ratio of Cox regression (risk ratio of rBPl2? placebo = 0.79, p = 0.13 for mortality or serious complication using "Present", p = 0.09 for mortality or serious complication using "Present or Clinically Present "that was stratified through the center). As a secondary analysis, the primary analysis was repeated with adjustment for significant covariates resulting in the following risk relationships: risk ratio of rBPl2? to placebo using "Present" = 0.79, p = 0.X4 (adjustment for age, source of X session, ISS'90 and units of PRBC by transfusion before Xa infusion of Xa drug); risk ratio using "Present or Clinically Present" = 0.8X, p = 0.16 (adjustment for age, ISS'90 and PRBC units of transfusion before infusion of the drug). The incidence of the event for each of the secondary efficacy measures evaluated is shown in Figure 1. Analysis of these secondary efficacy measures revealed lower frequencies of the following complications in patients treated with rBPl2? compared with patients treated with the placebo preparation: any complication, any serious complication, any abnormal function of the organ, any abnormal function of a serious organ, any infection, any serious infection and pneumonia. We observed slight reductions in favor of treatment with rBPl2? in Xa proportion of patients who developed disseminated intravascular coagulation or coaguolopathy, infection of the primary and secondary bloodstream and asymptomatic bacteriuria. Harmful events in this severely damaged population were frequent in patients treated with either rBPl2? or placebo. However, there were numerically higher percentages of patients with detrimental events in the placebo group compared to the rBP122 group. A higher percentage of patients were observed to experience any extremely abnormal post-treatment laboratory results in the placebo group compared to the group treated with rBPl2? - These data suggest a possible additional beneficial effect. In summary, this controlled clinical trial assessing a single-dose regimen has shown a trend in favor of treatment with rBPl2? in the primary endpoint of mortality or serious complication until Day 15. There were also reductions in the proportion of patients who experienced complications. These results, taken together, are compatible with the beneficial effect for treatment with rBPl2i in patients with hemorrhage due to trauma. It is to be expected that numerous modifications and variations of the invention described above occur to those skilled in the art. Accordingly, only the limitations appearing in the appended claims should be placed therein.

LIST OF SEQUENCES (1) GENERAL INFORMATION: (i) APPLICANTS: XOMA CORPORATION (ii) TITLE OF THE INVENTION: Therapeutic Uses of BPI Protein Products in Humans with Haemorrhage Due to Trauma (iii) NUMBER OF SEQUENCES: 2 (iv) ADDRESS FOR CORRESPONDENCE : (A) RECIPIENT: Marshall, O'Toole, Gerstein, Murray & Borun (B) STREET: 6300 Sears Tower, 233 South Wacker Drive (C) CITY: Chicago (D) STATE: Illinois (E) COUNTRY: United States of America (F) POSTAL CODE: 60606-6402 (v) READILY FORM COMPUTER: (A) MIDDLE TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: Patentln Résease # 1.0, Version # 1.25 (vi) DATA FROM THE CURRENT APPLICATION: (A) APPLICATION NUMBER: (B) SUBMISSION DATE: (C) CLASSIFICATION (vii) DATA FROM THE PREVIOUS APPLICATION: (A) APPLICATION NUMBER: 08 / 652,292 (B) SUBMISSION DATE: 23 May 1996 (C) CLASSIFICATION: (viii) ATTORNEY / AGENT INFORMATION: (A) NAME: Borun, Michael F. (B) REGISTRATION NUMBER: 25,447 (C) REFERRAL NUMBER / LAWYER'S OCA: 27129/33959 PCT (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 312 / 474-6300 (B) TELEFAX: 312 / 474-0448 (C) TELEX: 25-3856 (2) INFORMATION FOR SEQ ID NO: l: (i) CHARACTERISTICS OF THE SEQUENCE : (A) LENGTH: X8X3 base pairs (B) TYPE: nucleic acid (C) CHAIN TYPE: simple (D) TOPOLOGY: linear (Ü) TYPE OF MOLECULE: cDNA (ix) PARTICULARITY: (A) NAME / KEY : CDS (B) LOCATION: 31..1491 (ÍX) PARTICULARITY: (A) NAME / KEY: mat_peptide (B) LOCATION: 124..1491 (ix) PARTICULARITY: (A) NAME / KEY: misc_particularidad (D) ANOTHER INFORMATION: "rBPI" (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: l CAGGCCTTGA GGTTTTGGCA GCTCTGGAGG ATG AGA GAG AAC ATG GCC AGG GGC 54 Met Arg Glu Asn. Met Wing Arg Gly -31 -30 -25 CCT TGC AAC GCG CCG AGA TGG GTG TCC CTG ATG GTG CTC GTC GCC ATA 102 Pro Cys Asn Wing Pro Arg Trp Val Ser Leu Met Val Leu Val Wing -20 -15 -10 GGC ACC GCC GTG ACA GCG GCC GTC AAC CCT GGC GTC GTG GTC AGG ATC 1S0 Gly Thr Wing Val Thr Wing Wing Val Asn Pro Gly Val Val Val Arg lie -5 1 5 TCC CAG AAG GGC CTG GAC TAC GCC AGC CAG CAG GGG ACG GCC GCT CTG 198 Ser GIn Lys Gly Leu Asp Tyr Ala Ser Gln Gln Gly Thr Ala Ala Leu 10 15 20 25 CAG AAG GAG CTG AAG AGG ATC AAG ATT CCT GAC TAC TAC GAC AGC TTT 246 Gln Lvs Glu Leu Lys Arg lie Lys lie Pro Asp Tyr Ser Asp Ser Phe 30 35 40 AAG ATC AAG CAT CTT GGG AAG GGG CAT TAT AGC TTC TAC AGC ATG GAC 294 Lys He Lye Hie Leu Gly Lys Gly Hie Tyr Ser Phe Tyr Ser Met Asp 45 50 55 ATC CGT GAA TTC CAG CTT CCC AGT TCC CAG ATA AGC ATG GTG CCC AAT 342 He Arg Glu Phe Gln Leu Pro Ser Ser Gln He Ser Met Val Pro Asn 60 65 70 GTG GGC CTT AAG TTC TCC ATC AGC AAC GCC AAT ATC AAG ATC AGC GGG 390 Val Gly Leu Lys Phe Being As Asn Ala Asn He Lys He Ser Gly 75 80 85"AAA TGG AAG GCA CAA AAG AGA TTC TTA AAA ATG AGC GGC AAT TTT GAC 438 Lye Trp Lye Wing Gln Lys Arg Phe Leu Lys Met Ser Gly Asn Phe Asp 90 95 100 105 CTG AGC ATA GAA GGC ATG TCC ATT TCG GCT GAT CTG AAG CTG GGC AGT 486 Leu Ser He Glu Gly Met Ser Ser Wing Asp Leu Lye Leu Gly Ser 110 115 120 AAC CCC ACG TCA GGC AAG CCC ACC ATC ACC TGC TCC AGC TGC AGC AGC 534 Asn Pro Thr Ser Gly Lys Pro Thr He Thr Cye Ser Ser Cys Ser Ser 125 130 135 CÁC ATC AAC AGT GTC CAC GTG CAC ATC TCA AAG AGC AAA GTC GGG TGG 582 Hie He Aen Ser Val Hie Val Hie He Ser Lys Ser Lye Val Gly Trp 140 145 150 CTG ATC CAA CTC TC CAC AAA AAA ATT GAG TCT GCG CTT CGA AAC AAG 630 Leu He Gln Leu Phe His Lye Lye He Glu Be Ala Leu Arg Aen Lye 155 160 165 ATG AAC AGC CAG GTC TGC GAG AAA GTG ACC AAT TCT GTA TCC TCC AAG 678 Met Asn Ser Gln Val Cye Glu Lys Val Thr Aen Ser Val Ser Ser Lys 170 175 180 185 CTG CA CCT TAT TTC CAG ACT CTG CCA GTA ATG ACC AAA ATA GAT TCT 726 Leu C-ln Pro Tyr Phe Gln Thr Leu Pro Val Met Thr Lys He Asp Ser 190 195 200 GTG GGA ATC AAC TAT GGT CTG GTG GCA CCT CCA GCA ACC ACG GCT 774 Val Wing Gly He Aen Tyr Gly Leu Val Wing Pro Pro Wing Thr Thr Wing 205 210 215 GAG ACC CTG GAT GTA CAG ATG AAG GGG GAG TTT TAC AGT GAG AAC CAC 822 Glu Thr Leu Asp Val Gln Met Lys Gly Glu Phe Tyr Ser Glu Asn Hxe 220 225 230 CAC AAT CCA CCT CCC TTT GCT CCA CCA GTG ATG GAG TTT CCC GCT GCC 870 His Asn Pro Pro Pro Phe Pro Pro Wing Val Met Glu Phe Pro Wing Wing 235 240 245 CAT GAC CGC ATG GTA TAC CTG GGC CTC TAC GAC TTC TTC TAC AAC ACA 918 His Asp Arg Met Val Tyr Leu Gly Leu Ser Aep Tyr Phe Phe Asn Thr 250 255 260 265 GCC GGG CTT GTA TAC CAG GCT GCT GGG GTC TTG AAG ATG ACC CTT AGA 966 Wing Gly leu Val Tyr Gln Glu Wing Gly Val Leu Lys Met Thr Leu Arg 270 275 280 GAT GAC ATG ATT CCA AAG GAG TCC AAA TTT CGA CTG AC ACC AAG TTC 1014 Asp Asp Met He Pro Lye Glu Ser Lye Phe Arg Leu Thr Thr Lye Phe 285 290 295 TTT GGA ACC TC CTA CCT GAG GTG GCC AAG AAG TTT CCC AAC ATG AAG 1062 Phe Giy Thr Phe Leu Pro Glu Val Wing Lye Lye Phe Pro Aen Met Lys 300 305 310 ATA CAG ATC CAT GTC TCA GCC TCC ACC CCG CCA CAC CTG TCT GTG CAG 1110 He Gln He Hie Val Ser Wing Ser Thr Pro Pro Hie Leu Ser Val Gln 215 320 325 CCC ACC GGC CTT ACC TTC TAC TCC CCT GCC GTG GAT GTC CAG GCC TTT GCC 1158 Pro Thr Gly Leu Thr Phe Tyr Pro Wing Val Aep Val Gln Wing Phe Wing 330 335 340 345 GTC CTC CC AAC TCC TCC CTG GCT TCC CTC TTC CTG ATT GGC ATG CAC 1206 Val Leu Pro Aen Ser Ser Leu Wing Ser Leu Phe Leu He Gly Met Hie 350 355 360 ACA ACT GGT CC ATG GAG GTC AGC GCC GAG TCC AAC AGG CTT GTT GGA 1254 Thr Thr Gly Ser Met Glu Val Ser Wing Glu Ser Asn Arg Leu Val Gly 365 370 375 GAG CTC AAG CTG GAT AGG CTG CTC CTG GAA CTG AAG CAC TCA AAT ATT 1302 Glu Leu Lye Leu Aep Arg Leu Leu Leu Glu Leu Lys His Ser Aen He 380 385 390 GGC CCC TTC CCG GTT GAA TTG CTG CAG GAT ATC ATG AAC TAC ATT GTA 1350 Gly Pro Phe Pro Val Glu Leu Gluc Asp He Met Asn Tyr He Val 395 400 405 CCC ATT CTG GTG CTG CCC AGG GTT AAC GAG AAA CTA CAG AAA GGC TTC 1398 Pro He Leu Val Leu Pro Arg Val Asn Glu Lye Leu Gln Lye Gly Phe 410 415 420 425 CCT CTC CCG ACG CCG GCC AGA GTC CAG CTC TAC AAC GTA GTG CTT CAG 1446 'Pro Leu Pro Thr Pro Ala Arg Val Gln Leu Tyr Asn Val Val Leu Gln] 430 435 440 I CCT CAC CAG AAC TTC CTG CTG TTC GGT GCA GAC GTT GTC TAT AAA 1491; Pro His Gln Aen Phe Leu Leu Phe Gly Ala Asp Val Val Tyr Lye (445,450,455 i! TGAAGGCACC AGGGGTGCCG GGGGCTGTCA GCCGCACCTG TTCCTGATGG GCTGTGGGGC 1551 'ACCGGCTGCC TTTCCCCAGG GAATCCTCTC CAGATCTTAA CCAAGAGCCC CTTGCAAACT 1611 CTTCGACTC AGATTCAGAA ATGATCTAAA CACGAGGAAA CATTATTCAT TGGAAAAGTG 1671 \ CATGGTGTGT ATTTTAGGGA TTATGAGCTT CTTTCAAGGG CTAAGGCTGC AGAGATATTT i 1731 'CCTCCAGGAA TCGTGTTTCA ATTGTAACCA AGAAATTTCC ATTTGTGCTT CATGAAAAAA 1791 AACTTCTGGT TTTTTTCATG TG 1813 (2) INFORMATION FOR SEQ ID NO: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 487 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ü) TYPE OF MOLECULE: protein (xí) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 2 Met Arg Glu Asn Met Wing Arg Gly Pro Cys Aen Wing Pro Arg Trp Val 1-31 -30 -25 -20 Ser Leu Met Val Leu Val Wing He Gly Thr Wing Val Thr Ala Ala Val -15 -10 -5 1 Asn Pro Gly Val Val Val Arg He Ser Gln Lye Gly Leu Asp Tyr Ala s 10 15 Ser Gln Gln Gly Thr Ala Ala Leu Gln Lvs Glu Leu Lys Arg He Lye, 20 25 30 | He Pro Asp Tyr Ser Asp Ser Phe Lys He Lvs His Leu Gly Lvs Gly 35 40 45 Hie Tyr Ser Phe Tyr Ser Met Aep He Arg Glu Phe Glp Leu Pro Ser '50 55 60 65 Ser Glp He Ser Met Val Pro Aen Val Gly Leu Lys Phe Ser He Ser 70 75 80 Asn Wing Asn He Lys He Ser Gly Lys Trp Lys Wing Gln Lye Arg Phe 85 90 95 Leu Lys Met Ser Gly Aen Phe Asp Leu Ser He Glu Gly Met Ser He 100 105 110 Ser Wing Asp Leu Lye Leu Gly Ser Asn Pro Thr Ser Gly Lys Pro Thr 115 120 125 He Thr Cys Ser Ser Cys Ser Ser His He Aen Ser Val Hie Val Hie 130 135 140 145 He Ser Lye Ser Lye Val Gly Trp Leu He Gln Leu Phe Hie Lys Lye 150 155 160 He Glu Be Ala Leu Arg Asn Lye Met Asn Ser Gln Val Cye Glu Lye 165 170 175 Val Thr Asn Ser Val Ser Ser Lys Leu Gln Pro Tyr Phe Gln Thr Leu 180 185 190 Pro Val Met Thr Lye He Aep Ser Val Wing Gly He Aen Tyr Gly Leu 195 200 205 Val Wing Pro Pro Wing Thr Thr Wing Glu Thr Leu Asp Val Gln Met Lye 210 215 220 225 Gly Glu Phe Tyr Ser Glu Aen His His Asn Pro Pro Pro Phe Ala Pro 230 235 240 Pro Val Met Glu Phe Pro Ala Ala His Aep Arg Met Val Tyr Leu Glv 245 250 255 Leu Ser Aep Tyr Phe Phe Aen Thr Wing Gly Leu Val Tyr Gln Glu Wing 260 265 270 'Gly Val Leu Lys Met Thr Leu Arg Aep Aep Met He Pro Lye Glu Ser 275 280 285 Lys Phe Arg Leu Thr Thr Lys Phe Phe Gly Thr Phe Leu Pro Glu Val 290 295 300 305 Wing Lys Lys Phe Pro Asn Met Lye He Gln He His Val Ser Wing Ser 310 315 320 Thr Pro Pro His Leu Ser Val Gln Pro Thr Gly Leu Thr Phe Tyr Pro 325, 330 335 Wing Val Aep Val Gln Wing Phe Wing Val Leu Pro Asn Ser Ser Leu Wing 340 345? 350 Ser Leu Phe Leu He Gly Met Hie Thr Thr Thr Gly Ser Met Glu Val Ser 355 360 365 Wing Glu Ser Asn Arg Leu Val Gly Glu Leu Lye Leu Aep Arg Leu Leu 370 ^ s 375 380 385 Leu Glu Leu Lys His Ser Asn He Gly Pro Phe Pro Val Glu Leu Leu 390 395 400 Gln Asp He Met Asn Tyr He Val Pro He Leu Val Leu Pro Arg Val 405 410 415 Asn Glu Lys Leu Gln Lys Gly Phe Pro Leu Pro Thr Pro Wing Arg Val 420 425 430 Gln Leu Tyr Asn Val Val Leu Gln Pro His Gln Asn Phe Leu Leu Phe 435 440 445 Gly Ala Asp Val Val Tyr Lys 450 455

Claims (7)

CLAIMS:

1. The use of a protein product that increases bactericidal activity / permeability (BPI) in the preparation of a medicament for the treatment of a human being suffering from hemorrhage due to trauma.

2. The use of a protein product that increases bactericidal activity / permeability (BPI) in the preparation of a medicament for use in a method to treat or prevent a complication of abnormal lung function in a human being suffering from hemorrhage due to trauma

3. A use according to claim 1 or 2, wherein the protein product that increases the bactericidal activity / permeability is an ino-terminal fragment of the protein that increases bactericidal activity / permeability having a molecular weight of about 21 kD to 25 kD.

4. The use according to claim 1 or 2, wherein the protein product that increases the bactericidal activity / permeability is rBPl23 or a dimeric form thereof.

5. A use according to claim 1 or 2, wherein the protein product that increases Xa bactericidal activity / permeability is rBPl2 ?.

6. A use according to any of claims 1 to 5, wherein medicament is for administration together with a surfactant, a chemotherapeutic agent or an antimocrobial agent.

7. Use according to any of claims 1 to 5, wherein the mechanism is for administration in addition to the administration of at least two units of blood.