KR102369854B1 - A Contrast Agent Composition for Detecting Lesional Margin of Astrocytosis-Related Disease Comprising C11-Acetate as an Active Ingredient - Google Patents

Abstract

The present invention relates to a contrast agent composition for diagnosing astrocytosis, detecting reactive astrocytes, and obtaining images for brain tumor margins. The present invention provides image information on various diseases accompanied by excessive proliferation or activation of astrocytes, and in particular, by presenting the exact location of the dense area of these cells, the boundary between the lesion area and the surrounding normal area. Provides reliable information. Accordingly, the present invention can be particularly usefully used as a diagnostic image to guide surgical resection of a brain tumor in real time.

Description

Contrast Agent Composition for Detecting Lesional Margin of Astrocytosis-Related Disease Comprising C11-Acetate as an Active Ingredient comprising C11-acetate as an active ingredient

The present invention relates to a contrast agent composition for detecting or predicting the severity of a lesion boundary of an astrocytosis-related disease, specifically, a tumor boundary of a brain tumor, comprising C ¹¹ -acetate as an active ingredient.

PET (positron emission tomography) is one of the nuclear medicine testing methods using positron emission. It is a technology that injects a radioactive isotope that emits positrons into the body and then traces it and analyzes its distribution in the body. Clinical information and receptor images/metabolism images can be obtained. The positron used in PET has a positive charge while having physical properties similar to negatively charged electrons, and is emitted from radioactive isotopes such as C ¹¹ , N ¹³ , O ¹⁵ , and F ¹⁸ .

PET is an excellent contrast medium with the advantages of excellent sensitivity, high temporal resolution, and imaging of ^biological functions. can be detected. However, when diagnosing a brain tumor with F ¹⁸ -FDG, the tumor margin is not accurate, so it is difficult to use during surgery.

On the other hand, astrocytes are major cells that account for 30-50% of the total cell mass of the cerebral cortex, and in the nervous system, monitor the microenvironment of neurons in the central nervous system, transmit information, and form a blood-brain barrier to protect the brain. It has various functions such as protecting from the external environment and maintaining ion homeostasis. These astrocytes are activated and proliferate during brain damage and play an important role in repairing the damaged central nervous system. Astrocytes proliferate and undergo morphological changes during brain injury, and GFAP, an activation marker of astrocytes, increases. This condition is called astrocytosis or astrogliosis. Astrocytes are observed in various cranial nerve diseases such as dementia, inflammatory demyelinating diseases, acute traumatic brain injury, and neurodegenerative diseases such as Alzheimer's (McGraw et al, 2001; Krishnan et al., 2004). Therefore, when the change in the cellular environment in these brain diseases is clearly detected, it can be a reliable diagnostic marker, but an effective contrast agent for this has not yet been developed.

Numerous papers and patent documents are referenced throughout this specification and their citations are indicated. The disclosure contents of the cited papers and patent documents are incorporated herein by reference in their entirety to more clearly describe the level of the technical field to which the present invention pertains and the content of the present invention.

Non-Patent Document 1. C-11 radiochemistry in cancer imaging applications. Curr Top Med Chem. 10(11):1060-95(2010).

The present inventors made intensive research efforts to develop a reliable imaging technique for astrocytosis accompanied by significant proliferation and activation of astrocytes. As a result, ¹¹ C-acetate, a carbon isotope organic acid that has been limitedly applied as a contrast agent for the detection of specific cancer cells in which glucose metabolism is not active and a contrast agent for obtaining a cardiac image, has been specifically ^identified in the activated astrocytes. The present invention has been completed by discovering a new fact that stains are used for red blood cells and provides accurate information on dense regions of these cells.

Accordingly, an object of the present invention is to provide a contrast medium composition for obtaining a diagnostic image for astrocytosis.

Another object of the present invention is to provide a contrast agent composition for detecting reactive astrocytes.

Another object of the present invention is to provide a contrast medium composition for obtaining an image for a brain tumor margin.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a contrast medium composition for obtaining a diagnostic image for astrocytosis comprising ¹¹ C-acetate as an active ingredient.

The present inventors made intensive research efforts to develop a reliable imaging technique for astrocytosis accompanied by significant proliferation and activation of astrocytes. As a result, ¹¹ C-acetate, a carbon isotope organic acid that has been limitedly applied as a contrast agent for the detection of specific cancer cells in which glucose metabolism is not active and a contrast agent for obtaining a cardiac image, has been specifically ^identified in the activated astrocytes. A new finding was discovered that stained with red blood cells and provided accurate information about the dense regions of these cells. Above all, the present inventors first identified that ¹¹ C-acetate was specifically uptaken in reactive astrocytes, rather than ¹¹ C-acetate uptake by brain tumor cells as previously known.

¹¹ C-acetate used in the present invention may be in the form of various pharmaceutically acceptable ^{salts of 11 C} -acetic acid. As used herein, the term “pharmaceutically acceptable salt” includes salts derived from pharmaceutically acceptable inorganic acids, organic acids, or bases. Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, trifluoroacetic acid, citric acid, methane sulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals such as sodium, alkaline earth metals such as magnesium, and ammonium and the like. Specifically, the pharmaceutically acceptable salt used in the present invention is a sodium salt.

As used herein, the term “astrocytosis” refers to an abnormal increase in astrocytes due to damage to adjacent neurons, such as trauma of the central nervous system, tissue damage caused by tumors, infection, ischemia, stroke, neurodegenerative disease, etc. Or it is meant to encompass all pathological conditions that cause activation. Astrocytes that are activated or proliferated in lesions where brain tissue is damaged by various causes have some protective action on the regeneration process of brain tissue, but the increase in astrocytes appearing after injury inhibits the regeneration process by secreting various inflammatory substances. .

According to the present invention, when a diagnostic image is obtained by administering the composition of the present invention to a subject, the density or activity of abnormal astrocytes appears, and thus the onset of the disease as well as the precise lesion through the image of the region differentiated from the surrounding normal region. It is possible to obtain reliable information about the margin of the region.

According to a specific embodiment of the present invention, the astrocytosis is, for example, glioma, glioblastoma, astrocytoma, stroke, traumatic brain injury, epilepsy. (epilepsy), Rasmussen's syndrome, Alexander's disease, Amyotrophic Lateral Sclerosis (ALS), dementia due to degenerative brain disease, Parkinson's disease, inflammatory demyelinating disease demyelinating diseases) and Huntington's disease, but is not limited thereto, and includes any disease accompanying abnormal proliferation and/or activation of astrocytes.

The dementia caused by degenerative brain disease includes, for example, Alzheimer's disease (Alzheimer's disease), Lewy body dementia, and frontotemporal dementia, including, but not limited to, all dementias caused by degenerative brain disease.

More specifically, the glioma is a grade III or grade IV glioma.

As used herein, the term “administration” or “administering” refers to direct administration of an effective amount of the composition of the present invention to a subject so that the same amount is formed in the subject's body.

As used herein, the term “subject” includes, without limitation, a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, monkey, chimpanzee, baboon or rhesus monkey. Specifically, the subject of the present invention is a human.

As used herein, the term “effective amount” refers to an amount of an effective composition sufficient to provide information sufficient to make a clinical judgment on the subject's phenotype, metabolic activity, disease onset, and the area and boundary of the lesion when administered to a subject. means, and it can be expressed as a “diagnostic effective amount”.

As used herein, the term “diagnosis” refers to determination of a subject's susceptibility to a specific disease or disorder, determining whether a subject currently has a specific disease or disorder, and prognosis of a subject suffering from a specific disease or disorder. This includes monitoring the condition of a subject to provide information on the determination of treatment or treatment efficacy. The composition of the present invention shows not only the onset of a simple disease at the time of examination, but also the clear boundary and area of the lesion site and the process of its change with high temporal resolution, based on this, to determine the possibility of future tumor recurrence or metastasis, and furthermore, It can also be usefully used to evaluate therapeutic responsiveness.

According to a specific embodiment of the present invention, the image is a positron emission tomography (PET) image.

As used herein, the term “Positron Emission Tomography (PET)” refers to a radioactive isotope, such as C ¹¹ , N ¹³ , O ¹⁵ , and F ¹⁸ , which has a positive charge while having similar physical properties to electrons. It refers to an imaging method in which images are obtained by tracking positrons using a positron emission tomograph. Therefore, the composition of the present invention can be applied to all imaging methods including the principle of using positron emission, for example, positron/computed tomography (PET/CT) or magnetic material in which positron tomography and computed tomography are combined into one. It can also be applied to positron/magnetic resonance (PET/MRI) imaging in which positron emission factors and positron emission factors are combined.

According to a specific embodiment of the present invention, the composition of the present invention provides an image of the lesion margin of astrocytosis.

As used herein, the term “lesion margin” refers to a boundary between a two-dimensional or three-dimensional lesion area in vivo and a surrounding normal area, and specifically, it is distinguished by a clinician through a diagnostic image. possible boundaries. As will be described later, when the composition of the present invention is administered to a patient, the boundary of the astrocyte-dense region reflected by C ¹¹ acetate through the absorbed site, for example, the boundary of the tumor tissue of glioblastoma can be clearly indicated. there is. Through this, various medical actions that can be performed based on information on the exact location and area of the tumor, including surgical operations, can be performed more accurately. Therefore, in the present invention, the term “lesion margin” is used in the same sense as “excisional margin”.

According to another aspect of the present invention, the present invention provides a contrast agent composition for detecting reactive astrocytes comprising ¹¹ C-acetate as an active ingredient.

Since the contrast agent composition used in the present invention has already been described above, description thereof will be omitted to avoid excessive overlap.

As described above, ¹¹ C-acetate of the present invention is specifically absorbed into activated astrocytes in brain tissue damaged by various causes and provides an image thereof. Accordingly, the term reactive astrocyte is meant to include activated astrocytes.

According to another aspect of the present invention, the present invention provides a contrast medium composition for obtaining an image for a brain tumor margin (margin) comprising ¹¹ C-acetate as an active ingredient. The present invention can be used for image-guided surgery by providing an accurate image of the brain tumor margin as described above, and provides a more accurate tumor margin image as shown in the examples to be described later. It was confirmed that the patient's survival rate was significantly increased compared to the conventional MRI-based image-guided surgery. The brain tumor that can be imaged with the composition of the present invention may be a primary tumor or a metastatic tumor.

According to a specific embodiment of the present invention, the brain tumor is selected from the group consisting of glioma, glioblastoma and astrocytoma. More specifically, the glioma is a grade III or IV glioma.

According to another aspect of the present invention, the present invention provides a composition for predicting the severity of astrocytosis, excluding dementia caused by degenerative brain disease, comprising ¹¹ C-acetate as an active ingredient.

Since the contrast agent composition and astrocytosis used in the present invention have already been described above, description thereof will be omitted to avoid excessive overlap. As used herein, the term “severity” is meant to encompass disease progression, symptoms, degree of damage or risk of death in a patient suffering from a disease. As described above, the composition of the present invention provides quantitative information about the simple existence of reactive astrocytes or activated astrocytes as well as their density. Accordingly, the composition of the present invention can also provide accurate information on the severity of astrocytosis accompanied by significant proliferation and activation of astrocytes.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a contrast agent composition for diagnosing astrocytosis, detecting reactive astrocytes, and obtaining images for brain tumor margins.

(b) The present invention provides image information on various diseases accompanied by excessive proliferation or activation of astrocytes, and in particular, by presenting the exact location of the dense area of these cells, between the lesion area and the surrounding normal area It provides reliable information about the boundary part.

(c) Therefore, the present invention can be particularly usefully used as a diagnostic image for image-guided surgery in surgical resection of brain tumors, etc.

(d) In addition, the present invention shows not only the presence of a simple disease at the time of the examination, but also the process of changing the boundary and area of the lesion site with high temporal resolution, based on this, to detect the recurrence or metastasis of a future tumor early and further It can also be usefully used to evaluate therapeutic responsiveness to drugs.

1 is a picture showing a picture taken with C ¹¹ acetate PET in a mouse transplanted with glioblastoma cells.
FIG. 2 is a diagram showing a Kaplan-Meier curve in a C ¹¹ -acetate PET/MRI fusion image.
3 is a diagram showing C ¹¹ acetate PET (left) and MRI T1 contrast-enhanced images (right) in a patient with glioblastoma.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example

Animal Experiment - Imaging Experiment of Glioblastoma Mouse Model

Images were obtained with a PET scanner (Inveon, Siemens Healthcare, Erlangen, Germany) for mice (n=5) transplanted with Glioblastoma cells. For C ¹¹ acetate/PET imaging, mice were intravenously injected with C ¹¹ acetate (400 μCi/ 0.2ml Saline), incubated for 20 minutes, anesthetized with 2% (v/v) isoflurane, and acetate/PET data were recorded for 40 minutes. obtained. After image acquisition, the mice were observed until they woke up and placed in an isolation room for 24 hours to remove radiation disturbance. PET data were reconstructed into 2 repetitions and 18 subsets through 3D ordered subset expectation maximization (OSEM). The matrix size is 128 x 128 x 159 and the voxel size is 0.776 x 0.776 x 0.796 mm ³ . The standard uptake value (SUV) was calculated as tissue concentration (kBq/cc)/injection dose (kBq)/body weight (g).

As a result of photographing with C ¹¹ acetate PET, it was found that absorption was observed while forming a ring around the tumor ( FIG. 1 ).

PET imaging

All patients fasted at least 6 hours prior to ¹¹ C-acetate injection. 555 MBq (15 mCi) of ¹¹ C-acetate was intravenously injected into the patients, and an emission scan was performed 10 minutes later for 20 minutes. The patient's head was placed alongside the detector along the orbitometal line and then fixed. Images were acquired with a scout view of 10 mA and 120 kVp using a PET/CT scanner (Discovery 600, General Electric Medical Systems, Milwaukee, WI, USA), 0.8 s rotation time for attenuation correction, 60 mA, 120 kVp, Spiral CT scans were performed with a 3.75 mm cross-sectional thickness, 0.625 mm survey and 9.375 mm table feed per revolution. PET images were reconstructed using an iterative algorithm (iteration 2, subset 32).

Image transfer and fusion

Structural and metabolic image data and sequences were transferred from the image acquisition and communication system to a planning station via a local network for navigation surgery. A traditional navigation system (Stealth Treon, Medtronic, MN, USA) was used for surgery and image fusion, and contrast-enhanced T1-weighted MR images were used as reference images. Additional structural images, PET scans and MRI sequences, were automatically registered as reference images by the software.

For each patient's registration, at least five sticky reference skin markers were placed in a scatter pattern on the head region prior to image acquisition, and these markers were registered as pointers after the positions on these markers were defined in the 3D data set. Occasionally, a skin tracking method was used for patient enrollment. All procedures were performed in the same manner as in the traditional navigation system, and after registration, landmarks were repeatedly checked during the operation for accurate clinical application.

Use of image navigation during surgery

1) The position of the navigation pointer tip in the surgical area was marked on the workstation monitor with the corresponding position on the image. During resection of the tumor, surgeons often identified anatomical landmarks such as bone structures and cerebral cortex.

2) In the C ¹¹ -acetate PET/MRI fusion image-guided surgery group, the patient's tumor edge ^was determined directly from the MR image and the C ¹¹ -acetate PET fusion image. Only MR images were used.

Patient characteristics analysis before propensity score matching

The study was conducted on 58 glioma patients with IDH-mutant glioma and 82 IDH-wild-type glioma patients. There were no deaths among patients with IDH-mutant glioma and DH-wild-type diffuse astrocytoma ( FIG. 1 ). Among 80 patients with grade III and IV IDH-wild-type glioma, 37 patients underwent preoperative C ¹¹ -acetate PET irradiation and C ¹¹ -acetate PET/MRI fusion image-guided surgery. Forty-three patients who did not undergo C ¹¹ -acetate PET irradiation underwent MRI image-guided surgery.

Patient characteristics analysis after propensity score matching

In patients with matched propensity scores (n=48), no significant differences were observed between the two groups for age, sex, KPS, gross total resection, MGMT methylation, and Ki67 (all P> 0.05).

Kaplan-Meier analysis and Cox proportional hazards model for overall survival (OS) before propensity score matching

There were no deaths among patients with IDH-mutant glioma and grade II IDH-wild-type diffuse astrocytoma. Therefore, Kaplan-Meier analysis of OS was performed for grade III and IV IDH-wild-type gliomas. In the Kaplan-Meier curve, there was a significant difference in overall survival rate depending on whether C ¹¹ -acetate PET/MRI fusion image-guided surgery was performed ( FIG. 2 ). Specifically, the median overall survival was 22.37 (IQR 15.98-32.16) months in patients who underwent C ¹¹ -acetate PET/MRI fusion image-guided surgery, whereas 15.60 (IQR 8.93-) in patients who underwent MRI image-guided surgery. 23.43) months ( P=0.001 ).

In univariate analysis, age, C ¹¹ -acetate PET/MRI fusion image-guided surgery, and gross total resection were significant prognostic variables for overall survival. In multivariate analysis, age, C ¹¹ -acetate PET/MRI fusion image-guided surgery, and gross total resection were also statistically significant factors for overall survival (Table 1).

characteristic univariate P value multivariate P value gender 0.589 (0.330-1.050) 0.073 age 1.026 (1.005-1.049) 0.017 1.025 (1.002-1.049) 0.032 KPS 0.982 (0.962-1.003) 0.087 rank 1.920 (0.766-4.809) 0.164 acetate 2.332 (1.351-3.990) 0.002 1.829 (1.037-3.227) 0.037 Operation 2.017 (1.188-3.425) 0.009 2.026 (1.173-3.502) 0.011 MGMT 1.378 (0.808-2.347) 0.239 Ki67 1.012 (1.000-1.024) 0.053

Univariate hierarchical Cox regression for overall survival after propensity score matching

3 shows C ¹¹ acetate PET (left) and MRI T1 contrast-enhanced images (right), respectively, in patients with glioblastoma. It can be seen that yellow indicates the C ¹¹ acetate absorption boundary, purple indicates the contrast-enhanced portion of MRI, and acetate absorption is clearly located outside the MRI-enhanced portion.

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (13)

delete delete delete delete delete delete delete ¹¹ A contrast medium composition for obtaining an image of a brain tumor margin for image-guided surgery comprising C-acetate as an active ingredient.
The composition of claim 8, wherein the brain tumor is selected from the group consisting of glioma, glioblastoma and astrocytoma.
The composition of claim 8, wherein the contrast agent is a contrast agent for positron emission tomography (PET) images.
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