Treatment of Acute Tissue Injury

Abstract

The present invention is directed to the use of the compound PrC-210, or a pharma- ceutically acceptable acid addition salt thereof, for use in treating inflammation in a tissue of a subject, the inflammation resulting from acute tissue injury.

Description

[Technical field]

[0001] The present invention is directed to the use of the compound PrC-210, or a pharma- ceutically acceptable acid addition salt thereof, for use in treating inflammation in a tissue of a subject, the inflammation resulting from acute tissue injury. [Background technology]

[0002] End stage renal failure causes over 12 million deaths worldwide each year. The preferred treatment for patients with end stage renal disease is kidney transplantation. Over 90,000 kidney transplants are performed worldwide each year.

[0003] The transplantation process itself inflicts significant cellular and organotypic damage on the kidney, which impairs both short-term and long-term survival of the organ. There are four main types of damage that the kidney undergoes in allogeneic transplantation: (i) damage caused by reactive oxygen species (ROS) and reactive nitrogen species (RNS) before and during organ removal from the donor due to systemic shock and ischemia; (ii) damage caused by ROS or RNS during cold storage (cold ischemia); (iii) damage caused by ROS to the graft due to inflammation after allogeneic transplantation caused by innate immune responses and antibody-mediated rejection (ABMR); and (iv) damage caused by short-term or long-term immune responses to the transplanted organ (Sellares et al, Am J Transplant 2012;12:388-399).

[0004] Within 6 hours of reperfusion, neutrophils and macrophages infiltrate the allograft and stimulate resident dendritic cells to synthesize chemokines, which activate T lymphocytes and recruit acquired immune cells. When these immune cells infiltrate the proximal tubular epithelial cells, neutrophils produce myeloperoxidase and macrophages produce nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, which leads to local production of free radicals. This inflammatory process activates the complement pathway and promotes cellular remodeling and lysis in the renal allograft.

[0005] ABMR can be due to alloantibodies against the graft, de novo produced donor-specific antibodies (dnDSA), or both (Hidalgo et al, Am J Transplant 2009;9:2532-41).

[0006] Acute (minutes to days) inflammatory responses in renal allografts that transition to chronic (days to weeks) inflammatory responses result in the continuous production of free radicals (ROS and RNS) and inflammatory cytokines and chemokines, which can become severe and persistent and lead to long-term renal failure (Loupy et al; Nature Rev Nephrology; 2012;8:348-57).

[0007] To better understand the cellular, molecular, and immune pathways involved in the pathogenesis of inflammation and rejection in renal allografts, we established and characterized a rat model that recapitulates most of the clinical criteria for acute immune responses, ABMR, and kidney loss (Huang et al; Am J Transplant. 2014;14:1061-1072). We utilized this model to evaluate a number of novel post-allotransplant strategies.

[0008] Currently, there are two recognized approaches to reduce the severity of the acute and long-term immune response to the renal allograft: i) increasing the chances of finding a cross-matched donor, and ii) removing pre-existing antibodies against the renal allograft by desensitization (Stegall et al; Am J Transplant. 2006; 6:346-351). Summary of the Invention

[0009] The present invention is based on the finding that the severity of immune responses can be reduced (e.g., cellular damage in renal allografts) by using the free radical scavenger compound PrC-210 (3-(methylamino)-2-(methylaminomethyl)propane-1-thiol; Peebles et al; Radiat Res. 2012; 178: 57-68). PrC-210 directly and continuously scavenges and inactivates free radicals associated with (and causing) inflammatory genes, e.g., in newly transplanted renal allografts. This novel approach of the present invention will greatly advance the two existing strategies. The present invention itself may also generate new strategies to reduce acute damage in renal cells after transplantation.

[0010] PrC-210 is the prototype of a new family of direct acting small molecule aminothiol free radical scavengers. No side effects of nausea, vomiting, or hypotension have been reported with PrC-210 (Soref et al; Int J Rad Onc Biol Phys. 2012; 82: e701-707).

[0011] Two previous studies of renal transplantation in rodents have shown that PrC-210 suppresses free radical-induced kidney damage above background levels (i) after 30 hours of cold storage (Verhoven et al Transplantation Direct. 2020;6:e578) and (ii) during reperfusion injury during transplantation (Bath et al;Transplantation Direct. 2019;5:e549-555). This eliminates two essential factors that damage the transplanted kidney. The PrC-210 molecule has also been shown to suppress free radical-induced damage to various other organ environments. Thus, it is now clear that PrC-210 can protect allografts from oxidative stress caused by both cell-mediated and antibody-mediated rejection processes that generate free radicals.

[0012] Thus, one aspect of the invention is the compound PrC-210 (formula below), or a pharma- ceutically acceptable salt thereof, for use in the prophylaxis or treatment of inflammation in a tissue of a subject, the inflammation resulting from acute injury. [ka]

[0013] One aspect of the invention is the compound PrC-210 (formula below), or a pharma- ceutically acceptable salt thereof, for the manufacture of a medicament for preventing or treating inflammation in a tissue of a subject, the inflammation resulting from acute injury. [ka]

[0014] One aspect of the invention is a method of preventing or treating inflammation in tissue affected by acute injury, comprising administering to a subject the compound PrC-210 or a pharma- ceutically acceptable acid addition salt thereof. [ka]

[0015] One aspect of the invention is the compound PrC-210, or a pharma- ceutically acceptable acid addition salt thereof, for use in the prevention of organ transplant rejection. [ka]

[0016] One aspect of the present invention is the compound PrC-210 for use in the treatment of acute radiation syndrome (ARS).

[0017] One aspect of the present invention is a method of treating inflammation in a tissue of a subject, the inflammation resulting from acute tissue injury, by administering to the subject in need of treatment the compound PrC-210 or a pharma- ceutically acceptable acid addition salt thereof. [ka] [Brief description of the drawings]

[0018] [Figure 1] Schematic diagram of the experiment showing kidney surgery and administration of PrC-210. Kidneys from BN rats were transplanted into LEW rats. LT: left, RT: right, MTD: maximum tolerated dose, IP: intraperitoneal. [Diagram 2](A) Histological images of BN kidneys excised from BN rats. (B-D) Histological images of BN kidneys 20 hours after transplantation into LEW rats. Control kidneys (B, C) were washed with room temperature UW solution before transplantation. PrC-210 treated kidneys (D) were washed with room temperature UW solution containing 30 mM PrC-210 before transplantation. Recipient LEW rats were also given PrC-210 systemically via intraperitoneal administration three times within 8 hours after transplantation. The red arrow in panel B indicates the pathological image of tubulitis. The yellow arrow in panel C indicates the pathological image of pathological peritubular capillaritis. Panel E shows the sum of the severity scores of tubulitis and peritubular capillaritis. For each group, the scores are the sum of the scores in the three kidneys. [Diagram 3] (A-C) Histological images of kidneys 20 hours after transplantation. Sample images of renal tubules were randomly taken from the kidneys of each group in panels A-C. Next, H / E stained images (such as panels A-C in Figure 3) were analyzed using Image J. The number of pixels that were pink or blue above a threshold was counted, and the ratio was calculated and plotted. This allowed us to estimate the infiltration of leukocytes into the kidney. [Figure 4] The effect of PrC-210 administration on renal function 20 hours after transplantation of BN kidneys into LEW rats is shown. "20Hr" refers to rats that were not administered PrC-210. "20Hr+PrC-210" refers to rats that received three intraperitoneal injections of PrC-210 within 20 hours after transplantation. The recipient rats were then euthanized and serum was collected. p values ​​are shown. [Diagram 5] Figure 1 shows caspase activity in kidneys after transplantation. The activities of caspase 3 and caspase 7 activated by kidney supernatant were enzymatically measured. The reaction time was 60 min. [Figure 6] Figure 1 shows the effect of PrC-210 administration on cytokine levels in serum and kidney homogenates 20 hours after transplantation of BN kidneys into LEW rats. Levels of TIMP-1, TNFα and MIP-3A / CCL20 were determined by ELISA analysis. p values ​​are indicated. [Figure 7-1]A schematic diagram summarizing the efficacy of PrC-210 in suppressing organ damage following kidney transplantation. Three organs were examined in each group. [Figure 7-2] See item in Figure 7-1. [Figure 7-3] See item in Figure 7-1. [Figure 8] Schematic diagram summarizing the efficacy of PrC-210 in suppressing plasma caspase-1 in ICR mice, which were administered before or after a lethal dose of 8.68 Gy radiation. [Figure 9] Schematic diagram summarizing the efficacy of PrC-210 in suppressing caspase-8 in the brains of ICR mice, which were administered prior to a lethal dose of 8.68 Gy radiation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] In one embodiment of the invention, the compound PrC-210 is for use as described herein before or after an acute injury has occurred.

[0020] In another embodiment, the compound PrC-210 is for use in preventing the development of inflammation in tissues affected by acute injury.

[0021] In another embodiment of the present invention, the tissue affected by acute injury is selected from kidney tissue, liver tissue, heart tissue, brain / spinal or nerve tissue, blood vessels of the limbs, bone marrow tissue, intestinal tissue and lung tissue.

[0022] In another aspect of the invention, the tissue inflammation described herein is acute tissue inflammation.

[0023] In another aspect of the invention, the acute tissue inflammation described herein may be triggered by an acute traumatic event.

[0024] In another embodiment of the present invention, the acute tissue injury described herein is a secondary tissue injury.

[0025] In another embodiment of the present invention, the acute tissue injury described herein is fibrosis.Accordingly, one embodiment of the present invention is the compound PrC-210 for use in preventing or treating fibrosis caused by acute tissue injury.

[0026] In other aspects of the invention, the acute tissue damage described herein is present in a tissue that has been transplanted into a subject.

[0027] In another embodiment of the present invention, the acute tissue injury described herein occurs during a transplant procedure.

[0028] In another aspect of the invention, the acute tissue injury described herein results from surgery.

[0029] In another aspect of the invention, the acute tissue damage described herein results from trauma.

[0030] In another embodiment of the invention, the acute tissue damage described herein results from a burn or severe wound.

[0031] In another aspect of the invention, the acute tissue damage described herein results from a microbial or viral infection.

[0032] In other aspects of the invention, the acute tissue injury described herein results from stroke, myocardial infarction or pulmonary embolism.

[0033] In other aspects of the invention, the acute tissue injury described herein results from traumatic brain injury, spinal cord injury, or traumatic injury to a limb or central organ.

[0034] In other aspects of the invention, the acute tissue damage described herein is caused by radiation, e.g., the acute tissue damage is caused by acute radiation syndrome (ARS).

[0035] In other aspects of the invention, the acute tissue damage described herein results from ischemia or ischemia-reperfusion injury.

[0036] In another embodiment of the invention, the compound PrC-210 is administered systemically prior to, during, or after the tissue damage described herein has occurred.

[0037] In another embodiment of the invention, the compound PrC-210 is used prophylactically, either before or after the acute injury, in preventing secondary damage resulting from the acute injury.

[0038] In another embodiment of the invention, the compound PrC-210 is for use in fibrosis (pulmonary fibrosis, liver fibrosis, cardiac fibrosis, renal fibrosis, etc.).

[0039] In another embodiment of the invention, the compound PrC-210 is for use as described herein in the prevention of organ transplant rejection in a donor providing a transplanted organ.

[0040] In another embodiment of the invention, the compound PrC-210 is for use as described herein in a preservation solution for transporting donor organs.

[0041] In another embodiment of the invention, the compound PrC-210 is for use in a subject receiving a transplant organ from a transplant donor.

[0042] In another embodiment of the invention, the compound PrC-210 is for use in inhibiting organ damage associated with post-transplant inflammation.

[0043] According to the invention, the treatment reduces the histological score of tissue damage in exposed tissues, which may be selected from kidney tissue, bone marrow tissue, brain and spinal cord tissue, cardiac tissue, muscle tissue, nervous tissue, jejunal tissue and lung tissue.

[0044] Further according to the invention, the treatment reduces the levels of inflammatory markers, which may be selected from TNFα in serum, NOS2, albumin α in serum, infiltration of mononuclear cells into injured tissue, and chemokine synthesis in resident dendritic cells.

[0045] The present invention also relates to treatments that reduce or normalize elevated levels of activated caspases (such as caspase 1, caspase 3 / 7, caspase 8, etc.) to physiological levels.

[0046] The present invention also relates to one or more of the following treatments: (i) upregulating levels of MIP-3a / CCL20; (ii) modulating and activating T cells, particularly T cells; and (iii) reducing levels of tissue inhibitor of metalloproteinase-1 (TIMP-1).

[0047] According to the present invention, acute inflammation is reduced.

[0048] The present invention also normalizes inflammation-associated cell death.

[0049] The present invention also reduces secondary tissue damage and fibrosis.

[0050] According to the present invention, the tissue to be treated is a tissue transplanted into a subject, and the damage is related to the transplant. The damage is due to acute irradiation or a nuclear event. The subject has undergone surgery resulting from tissue damage. The subject has suffered trauma resulting from tissue damage. The subject has suffered a burn or a severe wound resulting from tissue damage. The tissue damage is due to a microbial or viral infection. The tissue damage is due to a stroke, myocardial infarction or pulmonary embolism. The tissue damage is due to a traumatic brain injury, spinal cord injury or traumatic injury of a limb or central organ. Alternatively, the tissue damage is due to ischemia or ischemic reperfusion injury.

[0051] In one aspect of the present invention, the compound PrC-210 described herein is used for treating acute radiation syndrome (ARS).The compound PrC-210 described herein may also be used for preventing acute radiation syndrome (ARS).For example, the compound PrC-210 described herein may be administered to a subject who is going to undergo radiotherapy.That is, the compound PrC-210 may be administered to a subject prior to undergoing radiotherapy.

[0052] The invention may involve systemic administration of an effective amount of a compound at an effective time, which may be before, during or after a damaging event that causes tissue damage. In one aspect, the compound is administered prophylactically to protect tissue.

[0053] As used herein, the term "inflammation in a tissue" refers to inflammation resulting from a local immune, vascular, or inflammatory cell response to tissue injury or infection.

[0054] As used herein, the term "tissue" refers to a structure of similar types of specialized cells. Tissues perform specific functions within the body of multicellular organisms (connective tissue, epithelial tissue, muscle tissue, nervous tissue, etc.). Epithelial tissues are located on the surface and in the cavities of the body and protect the body from dehydration and mechanical damage. Connective tissues connect and bind separate tissues and organs. Connective tissues are composed of a small number of cells embedded in a matrix secreted by connective tissue cells. Muscle tissues are involved in controlling internal functions (digestion, respiration, urination, etc.). Muscle tissues are also involved in controlling external functions (movement of body parts, etc.). Nerve tissues coordinate internal and external functions through various stimuli.

[0055] As used herein, the term "inflammation caused by acute injury" refers to inflammation caused by acute damage to the body, which may occur in any organ of the human body that has been subjected to trauma or physical damage.

[0056] As used herein, the term "organ" refers to any organ in the human body, including, by way of example, the kidney, bone marrow, jejunum (small intestine), colon, lung, liver, and brain.

[0057] As used herein, the term "organ tissue" refers to any organ tissue of the human body, including, by way of example, kidney tissue, bone marrow tissue, jejunum (small intestine) tissue, colon tissue, lung tissue, liver tissue, and brain tissue.

[0058] As used herein, the term "secondary tissue damage" refers to destructive and self-replicating biological changes in cells and tissues. Within hours to days after the initial damage (primary damage), cells or tissues become dysfunctional or die. In many cases, the initial damage is mechanical.

[0059] As used herein, the term "fibrosis" refers to the development of fibrous connective tissue in response to repeated injury or damage. Fibrosis may be referred to as connective tissue deposition when it occurs during natural healing, or as excess tissue deposition when it occurs during a pathological process. When fibrosis occurs in response to injury, the term "scar" is used. Examples of fibrosis in the human body include pulmonary fibrosis, liver fibrosis (liver cirrhosis), cardiac fibrosis (cardiac fibrosis), mediastinal fibrosis, retroperitoneal fibrosis, bone marrow fibrosis, skin fibrosis, scleroderma, and systemic sclerosis.

[0060] Pulmonary fibrosis can result from long-term tuberculosis or pneumonia infection. The disease can also result from exposure to occupational hazards such as coal dust, or from the genetic cause of cystic fibrosis.

[0061] Liver fibrosis or cirrhosis refers to scar tissue and nodules that replace liver tissue and disrupt liver function. The disease is often due to alcoholism, fatty liver, hepatitis B or C.

[0062] Cardiac fibrosis or cardiac fibrosis refers to areas of the heart damaged by myocardial infarction, which may progress to fibrosis.

[0063] Mediastinal fibrosis is a type of fibrosis characterized by calcified fibrosis of lymph nodes, which may lead to obstruction of arteries or blood vessels.

[0064] Retroperitoneal fibrosis is the fibrosis of the soft tissues of the retroperitoneum, which contains the aorta, kidneys, and many other structures.

[0065] Myelofibrosis is the scarring of the bone marrow, which prevents it from producing normal blood cells.

[0066] Dermal fibrosis is scar tissue that forms in the skin in response to injury, also called keloids.

[0067] Scleroderma, or systemic sclerosis, is an autoimmune disease of connective tissue that initially affects the skin but can also involve other organs (such as the kidneys, heart, and lungs).

[0068] The term "acute radiation syndrome (ARS)" refers to a set of health effects resulting from exposure to large amounts of ionizing radiation over a short period of time, also known as radiation sickness or radiation poisoning. An example of radiation poisoning is radiation resulting from a nuclear event.

[0069] As used herein, the term "transplanted tissue" or "transplant organ" refers to a tissue or organ that is removed from a subject (donor) and transplanted into a patient in need of the tissue or organ.

[0070] As used herein, the term "allograft" refers to the surgical transplantation of tissue (or organ) between genetically distinct individuals of the same species.

[0071] As used herein, the term "prophylactic therapy" refers to the use of compound PrC-210 or its pharma- ceutically acceptable acid addition salt to prevent inflammation in tissue as described herein.This term also includes the prevention of fibrosis as described herein.This term also includes the prevention of rejection of transplanted organ in subjects who receive organs from transplant donors.

[0072] As used herein, the term "suppression of inflammation-associated organ damage after transplantation" refers to the use of the compound PrC-210 to prevent or reduce inflammation in an organ in a patient who has received an organ from a donor.

[0073] As used herein, the term "compound PrC-210" refers to the following compound or a pharma- ceutically acceptable acid addition salt thereof: This compound and its method of preparation are disclosed, inter alia, in U.S. Patent No. 7,314,959. [ka] EXAMPLES

[0074] Example 1 To investigate the efficacy of PrC-210 in inhibiting inflammation-induced damage in allografted organs, we used a rat kidney allograft model. In this model, ischemia and reperfusion at the time of transplantation are largely suppressed, and only tertiary inflammation occurs due to the kidney allograft. In this experiment, kidneys were transplanted from donor Brown strain (BN) to recipient Lewis strain (LEW) rats (Fig. 1). PrC-210 was administered before and after transplantation to examine whether it has a protective effect in inhibiting any inflammation-related damage associated with transplantation of BN kidneys into LEW rats. BN rat kidneys were flushed with UW solution containing PrC-210 and then immediately transplanted into syngeneic recipient LEW rats. Virtually no ischemia occurred. PrC-210 was administered systemically to recipient rats immediately after kidney transplantation and 8 hours after kidney transplantation. The dose was sufficient to continuously scavenge free radicals in the transplanted kidney. Twenty hours after transplantation, transplanted kidneys and plasma were harvested to measure the ability of PrC-210 to (i) suppress inflammatory by-products and (ii) protect the kidney.

[0075] Example 2 Histological images of transplanted BN kidneys 20 hours after transplantation are shown (Figure 2, Panels A-D). In tissues treated with UW solution alone, the Banff scores of tubulitis and peritubular capillitis were clearly elevated (red and yellow arrows). The total pathology score is shown in Panel E. When BN kidneys were perfused with PrC-210-containing UW solution and further administered intraperitoneally after transplantation, inflammatory pathology in the kidney was clearly suppressed (Figure 2, Panel D). At this time, the Banff scores of tubulitis and peritubular capillitis were suppressed to the same level as in the control BN group (0 Hr) (Figure 2, Panel E).

[0076] Example 3 Histological images of BN kidneys washed with UW solution alone (20 Hr) showed a marked reduction in the thickness of the tubular brush border epithelium compared to control BN kidneys (0 Hr) immediately after removal from BN rats (Figure 3, Panels A-B). Histological images of BN kidneys administered PrC-210 by perfusion with UW solution and intraperitoneal injection clearly maintained the integrity of the tubular brush border (Figure 3, Panel C).

[0077] The same blinded tissue sections were examined for mononuclear leukocyte infiltration. 20 hours after washing with UW solution alone, the number of blue nuclei was significantly increased (scored as blue pixels) in BN kidneys, reflecting mononuclear leukocyte infiltration. On the other hand, BN kidneys washed and treated with PrC-210 showed significantly suppressed mononuclear infiltration compared to untreated BN kidneys (p=0.011). At this time, the inflammatory infiltration score was statistically the same as that of the control group (0Hr).

[0078] Example 4 Serum creatinine and serum BUN were measured to evaluate the function of the BN kidney 20 hours after allograft transplantation (Figure 4). Administration of PrC-210 significantly reduced creatinine and BUN, which are markers of kidney damage (p=0.032 and p=0.046, respectively).

[0079] Example 5 In BN kidney allografts not treated with PrC-210, the level of activated caspases in kidney homogenates was significantly reduced 20 hours after transplantation (Fig. 5). In a system in which BN kidneys perfused with PrC-210-containing UW solution were transplanted into Lewis rats to which PrC-210 had been administered systemically, the activity of activated caspases was comparable to that of kidneys in the control group (0 Hr). This result indicates that damage to the kidney after kidney transplantation without ischemia is reduced.

[0080] Example 6 Renal homogenate supernatants from the control group at 0 hours and the untreated group at 20 hours were screened to detect changes in the expression levels of inflammation-related cytokines and chemokines 20 hours after transplantation. Proteome Profiler 29 cytokine array was used for screening. As shown in the insets of the two microarrays in Figure 6, changes were observed in TIMP-1, TNFα, and MIP-3a / CCL20. The changes were then quantified in the kidney homogenate and serum, respectively, using ELISA plates. In this experiment, rats treated with PrC-210 were also included. At 20 hours after transplantation, the levels of both TIMP-1 and TNFα were elevated. However, in the presence of PrC-210, both levels were reduced (Figure 6, Panels A-C). At 20 hours, the levels of MIP-3a / CCL20 were elevated, but significantly elevated in the rats treated with PrC-210 (Figure 6, Panel D).

[0081] The efficacy of PrC-210 in preventing organ damage following kidney transplantation is summarized in Figure 7. Three organs were examined in each group.

[0082] Example 7 Figure 8 shows that plasma caspase-1 is suppressed by PrC-210 treatment for 6 days after lethal radiation, supporting the long-term inhibition of the inflammasome. Caspase-1 is a central activator of acute inflammation in neutrophil cells. It recruits T cells and monocytes and activates apoptosis.

[0083] Example 8 Inhibition of caspase-8 in the brain after acute injury (e.g., radiation) is supported by the decreased activity of TNFα and the upregulation of TRL and FAS receptors in the brain (Figure 9). These three biomarkers are upregulated after acute injury (e.g., radiation, traumatic brain injury), which leads to the upregulation of caspase-8. This supports that inflammation, such as microglial activation, is normalized by administration of PrC-210. The effect of caspase-8 continues for more than 6 days. This supports that PrC-210 inhibits inflammation for a long period of time.

Claims

1. A pharmaceutical preparation for treating inflammation in the tissue of a subject, comprising the compound PrC-210 or a pharmaceutically acceptable acid addition salt thereof, wherein the inflammation is caused by secondary tissue damage, pharmaceutical preparation. 【Chemical 1】

2. The tissue invaded by secondary tissue damage is selected from renal tissue, hepatic tissue, cardiac tissue, cerebral tissue, spinal tissue, nerve tissue, vascular tissue of the extremities, bone marrow tissue, intestinal tissue (jejunal tissue) and lung tissue, The pharmaceutical preparation according to Claim 1.

3. The secondary tissue damage has fibrosis, The pharmaceutical preparation according to Claim 1 or 2.

4. The secondary tissue damage is in the tissue transplanted into the subject, The pharmaceutical preparation according to Claim 1.

5. The secondary tissue damage is caused by surgery, The pharmaceutical preparation according to Claim 1.

6. The secondary tissue damage is caused by trauma, The pharmaceutical preparation according to Claim 1.

7. The secondary tissue damage is caused by burns or severe trauma, The pharmaceutical preparation according to Claim 1.

8. The secondary tissue damage is caused by microbial infection or viral infection, The pharmaceutical preparation according to Claim 1.

9. The secondary tissue damage is caused by traumatic brain injury, spinal cord injury, or traumatic injury to the extremities or central organs, The pharmaceutical preparation according to Claim 1.

10. The compound PrC-210 is administered systemically before the secondary tissue damage occurs, when the secondary tissue damage occurs, or after the secondary tissue damage occurs, The pharmaceutical preparation according to Claim 1.

11. The fibrosis is pulmonary fibrosis, hepatic fibrosis, cardiac fibrosis, renal fibrosis, mediastinal fibrosis, retroperitoneal fibrosis, bone marrow fibrosis, skin fibrosis, scleroderma or systemic scleroderma, The pharmaceutical preparation according to Claim 3.

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