WO2022186335A1

WO2022186335A1 - Side effect prediction method and device for same

Info

Publication number: WO2022186335A1
Application number: PCT/JP2022/009132
Authority: WO
Inventors: 秀徳高橋
Original assignee: 学校法人自治医科大学
Priority date: 2021-03-03
Filing date: 2022-03-03
Publication date: 2022-09-09
Also published as: JPWO2022186335A1

Abstract

The present disclosure relates to estimating the likelihood that a subject will develop side effects such as uveitis after administration of brolucizumab. This method involves using the concentration of at least one protein and/or the proportion of Th2 cells among the CD4-positive lymphocytes in a body fluid such as anterior aqueous humor, serum, or blood that has been isolated from a subject before administration of brolucizumab as an indicator to estimate the likelihood that the subject will develop side effects after administration of brolucizumab.

Description

Side effect prediction method and device therefor

Cross-reference to related applications

This application is an application that enjoys the benefit of the priority of Japanese Patent Application No. 2021-33555 (filing date: March 3, 2021), which is incorporated herein by reference in its entirety.

The present invention relates to a method for estimating the possibility of developing side effects and a kit, device and program used therefor, more specifically, a method for estimating the possibility of developing endophthalmitis occurring after administration of brolucizumab and a kit, device and program used therefor, The present invention relates to a method, device and program for assisting in determining whether or not to administer brolucizumab, and to a method, device and program for assisting in determining whether or not an anti-inflammatory agent is required when administering brolucizumab.

Brolucizumab is a recently approved anti-vascular endothelial growth factor (VEGF) drug, a humanized single-chain variable fragment (scFV) consisting of antibody variable domains involved in target binding (Non-Patent Document 1). Brolucizumab is highly soluble, allowing the administration of concentrated, high concentrations of the molecule intravitreally. Brolucizumab also has improved tissue penetration and bioavailability due to its small molecular size, which may lead to longer durability and improved efficacy in the treatment of neovascular-associated age-related macular degeneration (AMD). (Non-Patent Document 2). However, in its phase III trials, the HAWK and HARRIER trials, higher rates of aseptic endophthalmitis (IOI), retinal vasculitis, and retinal vascular occlusion after injection of brolucizumab compared with aflibercept was observed in (Non-Patent Document 3). In addition, several severe cases such as central retinal artery occlusion-like findings have been reported after market launch (Non-Patent Document 4). Severe side effects have been reported to occur weeks after several injections of brolucizumab, so IOIs have been hypothesized to be due to delayed-type hypersensitivity.

To date, no research has confirmed the cause of IOI, and the challenge is to understand its risk factors and the underlying mechanism of IOI. In particular, it is desired to predict the possibility of developing endophthalmitis after administration of brolucizumab.

The present inventors investigated subjects with age-related macular degeneration (AMD) who received brolucizumab injections, and compared eyes that developed endophthalmitis within 3 months after the first brolucizumab injection and eyes that did not, respectively. As a result of examining the difference in the protein profile of the anterior aqueous humor immediately before injection, we found that patients who developed endophthalmitis after brolucizumab injection had a characteristic profile of specific protein concentrations in the anterior aqueous humor before injection. rice field. In addition, blood was collected after that, and it was found that a characteristic profile was observed in serum protein concentration, and that the proportion of Th2 cells in blood CD4-positive lymphocytes was significantly small. The present invention is based on such findings, and more specific aspects relate to the following matters.

[a1] A method for estimating the possibility of developing a side effect after administration of brolucizumab in a subject, comprising at least one protein concentration and/or CD4-positive lymphocyte concentration in body fluids isolated from the subject before administration of brolucizumab A method using the ratio of Th2 cells as an index.
[a2] The method of [a1], wherein the side effect is a side effect associated with intraocular inflammation.
[a3] The method of [a1] or [a2], wherein the side effect is non-infectious endophthalmitis.
[a4] Any of [a1] to [a3], wherein the side effect is selected from the group consisting of uveitis, retinal vasculitis, iritis, anterior chamber inflammation, vitreous inflammation, and iridocyclitis The method described in item 1.
[a5] Any one of [a1] to [a4], wherein the body fluid is selected from the group consisting of blood, plasma, serum, saliva, urine, sweat, tears, vitreous humor, anterior aqueous humor, and posterior aqueous humor The method described in the section.
[a6] at least the protein is selected from the group consisting of MCP-1, IP-10, P-selectin, CXCL1, VEGF-A, IL-6, MMP-9, IL-1β, CCL7, and G-CSF The method of any one of [a1]-[a5], comprising one protein.
[a7] Low MCP-1, high IP-10, high P-selectin, high CXCL1, high VEGF-A, low IL-6, high MMP-9, high IL-1β, low The method of any one of [a1] to [a6], wherein the index is at least one characteristic selected from the group consisting of CCL7, low G-CSF, and a low percentage of Th2 cells among CD4-positive lymphocytes. .
[a8] the subject has at least one of age-related macular degeneration (AMD), diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity, [a1] The method according to any one of to [a7].
[a9] The method of any one of [a1]-[a8], wherein the subject has been administered aflibercept prior to administration of brolucizumab.
[a10] A method for estimating the likelihood of developing endophthalmitis in a subject after administration of brolucizumab, comprising:
Low MCP-1, high IP-10, high P-selectin, high CXCL1, high VEGF-A in anterior aqueous humor and/or low IL-6, high MMP-9 in serum before brolucizumab administration , and high IL-1β concentrations as indicators,
Subject has at least one of age-related macular degeneration (AMD), diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity, [a1]-[a9 ] The method according to any one of the above.
[a11] The following formula:
a×log ₁₀ (MCP-1)＋b×log ₁₀ (IP-10)＋c×log ₁₀ (P-selectin)＋d×log ₁₀ (CXCL1)＋e×log ₁₀ (VEGF-A)＋f＞0
(In the formula, MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A each represent the protein concentration (pg/mL) in body fluids, a = -11 to -0.91, b = 0.50 to 5.2, c = 0.66 to 4.6, d = -0.17 to 4.2, e = 0.0071 to 2.3, and f = -12 to 11)
and/or a × log ₁₀ (IL-6) + b × log ₁₀ (MMP-9) + c × log ₁₀ (IL-1β) + d > 0
(In the formula, IL-6, MMP-9, and IL-1β each represent the serum protein concentration (pg/mL), a = -35 to -12, b = 1.7 to 4.7, c = 7.1 to 30, and d = -17 to 1.2)
and/or blood Th2 < a
(where a = 2.5 to 4.6)
and/or a x Th2 + b x _log10 (MMP-9) + c x _log10 (P-selectin) + d x _log10 (CCL7) + e x _log10 (G-CSF) + f > 0
(where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, MMP-9, P-selectin, CCL7, and G-CSF each represent protein concentration in serum (pg/mL), a = -1.6 to -0.78, b = 4.2 to 9.6, c = 12 to 31, d = -76 to -27, e = -19 to -5.3, and f = -12 to 40)
The method according to any one of [a1] to [a10], which estimates the possibility of developing a side effect due to
[a12] The following formula:
-5.8 x log ₁₀ (MCP-1) + 2.8 x log ₁₀ (IP-10) + 2.6 x log ₁₀ (P-selectin) + 2.0 x log ₁₀ (CXCL1) + 1.1 x log ₁₀ (VEGF- A) - 0.12 > 0
(wherein MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A each represent the protein concentration in the anterior aqueous humor (pg/mL))
and/or −23×log ₁₀ (IL-6) + 3.2×log ₁₀ (MMP-9) +19×log ₁₀ (IL-1β)) −8.1 >0
(wherein IL-6, MMP-9, and IL-1β each represent protein concentration in serum (pg/mL))
and/or blood Th2 < 3.4
and/or -1.2 x Th2 + 6.9 x _log10 (MMP-9) + 22 x _log10 (P-selectin) -52 x _log10 (CCL7) -12 x _log10 (G-CSF) +14 > 0
(Where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, and MMP-9, P-selectin, CCL7, and G-CSF are protein concentrations in serum (pg/mL), respectively.)
The method according to any one of [a1] to [a11], which estimates the possibility of developing endophthalmitis due to
[a13] A method for estimating the likelihood of developing endophthalmitis in a subject after administration of brolucizumab, comprising:
Concentrations of MCP-1, IP-10, P-selectin, CXCL1, VEGF-A in anterior aqueous humor and/or IL-6, MMP-9, and IL in serum isolated from subjects prior to administration of brolucizumab measuring the concentration of -1β,
Based on measured levels of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A and/or levels of IL-6, MMP-9, and IL-1β, endophthalmitis develops The method according to any one of [a1]-[a12], comprising the step of estimating likelihood.
[a14] A kit for use in the method according to any one of [a1] to [a13] above,
A kit comprising a capture antibody for an indicator protein immobilized on a solid phase support, and a detection antibody for the indicator protein.
[a15] The indicator protein is
The kit of [a14] selected from MCP-1, IP-10, P-selectin, CXCL1, VEGF-A, IL-6, MMP-9, IL-1β, CCL7 and G-CSF.
[a16] A device for estimating the likelihood of developing a side effect in a subject after administration of brolucizumab, comprising:
an information acquisition unit that acquires information on the percentage of Th2 cells in at least one protein concentration and/or CD4-positive lymphocytes in body fluid before administration of brolucizumab;
An apparatus comprising an estimation unit for estimating the possibility of developing a side effect based on protein concentration and/or information on the ratio of Th2 cells in CD4-positive lymphocytes, and a notification unit for outputting the estimation result.
[a17] is estimated by the following formula:
-5.8 x log ₁₀ (MCP-1) + 2.8 x log ₁₀ (IP-10) + 2.6 x log ₁₀ (P-selectin) + 2.0 x log ₁₀ (CXCL1) + 1.1 x log ₁₀ (VEGF- A) - 0.12 > 0
(wherein MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A each represent the protein concentration in the anterior aqueous humor (pg/mL))
and/or −23×log ₁₀ (IL-6) + 3.2×log ₁₀ (MMP-9) +19×log ₁₀ (IL-1β)) −8.1 >0
(wherein IL-6, MMP-9, and IL-1β each represent protein concentration in serum (pg/mL))
and/or blood Th2 < 3.4
and/or -1.2 x Th2 + 6.9 x _log10 (MMP-9) + 22 x _log10 (P-selectin) -52 x _log10 (CCL7) -12 x _log10 (G-CSF) +14 > 0
(Where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, and MMP-9, P-selectin, CCL7, and G-CSF are protein concentrations in serum (pg/mL), respectively.)
The apparatus according to [a16], which is carried out based on.
[a18] A program for estimating the likelihood of developing a side effect after administration of brolucizumab in a subject, comprising:
Obtaining information on the concentration of at least one protein in the body fluid and/or the percentage of Th2 cells in the CD4-positive lymphocytes before administration of brolucizumab;
A step of estimating the possibility of developing side effects based on information on the protein concentration and/or the ratio of Th2 cells in CD4-positive lymphocytes,
A program that executes a process of outputting an estimation result.
[a19] is estimated by the following formula:
-5.8 x log ₁₀ (MCP-1) + 2.8 x log ₁₀ (IP-10) + 2.6 x log ₁₀ (P-selectin) + 2.0 x log ₁₀ (CXCL1) + 1.1 x log ₁₀ (VEGF- A) - 0.12 > 0
(wherein MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 each represent the protein concentration in the anterior aqueous humor (pg/mL))
and/or −23×log ₁₀ (IL-6) + 3.2×log ₁₀ (MMP-9) +19×log ₁₀ (IL-1β)) −8.1 >0
(wherein IL-6, MMP-9, and IL-1β each represent protein concentration in serum (pg/mL))
and/or -1.2 x Th2 + 6.9 x _log10 (MMP-9) + 22 x _log10 (P-selectin) -52 x _log10 (CCL7) -12 x _log10 (G-CSF) +14 > 0
(Where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, and MMP-9, P-selectin, CCL7, and G-CSF are protein concentrations in serum (pg/mL), respectively.)
and/or blood Th2 < 3.4
The program according to [a18], which is carried out based on
[a20] A method for assisting in determining whether or not to administer brolucizumab to a subject, comprising at least one protein concentration and/or Th2 cells in CD4-positive lymphocytes in body fluid prior to administration of brolucizumab isolated from the subject A method that uses the ratio of
[a21] A method for assisting in determining whether concomitant use of an anti-inflammatory agent is required when administering brolucizumab to a subject, comprising: A method in which the ratio of Th2 cells in positive lymphocytes is used as an index.
[a22] A method for estimating the possibility of developing a side effect after administration of brolucizumab in a subject, wherein the Th1/Th2 ratio in blood isolated from the subject before administration of brolucizumab is used as an index.
[a23] A computer-readable storage medium non-transitoryly storing a program for causing a computer to execute a process including the following steps:
Obtaining information on the concentration of at least one protein in the body fluid and/or the percentage of Th2 cells in the CD4-positive lymphocytes before administration of brolucizumab;
A step of estimating the possibility of developing side effects based on information on the protein concentration and/or the ratio of Th2 cells in CD4-positive lymphocytes,
A step of outputting the estimation result.
[a24] A computer-implemented method for estimating the likelihood of developing a side effect in a subject after administration of brolucizumab, comprising:
Obtaining information on the concentration of at least one protein in the body fluid and/or the percentage of Th2 cells in the CD4-positive lymphocytes before administration of brolucizumab;
A step of estimating the possibility of developing side effects based on information on the protein concentration and/or the ratio of Th2 cells in CD4-positive lymphocytes,
A method including the step of outputting an estimation result.
[a25] The method of [a1], wherein serum levels of IL-6, MMP-9, and IL-1β are used as indicators.
[a26] A method for estimating the possibility of developing a side effect in a subject after administration of brolucizumab, wherein the rate of Th2 cells in the blood of the subject is used as an index.
[a27] The method of [a26], wherein a small proportion of Th2 cells in CD4-positive lymphocytes is used as an indicator.
[a28] The following formula:
Blood Th2 < a (where a = 2.5 to 4.6)
The method of [a26], which estimates the possibility of developing a side effect due to
[a29] A method for estimating the likelihood of developing endophthalmitis in a subject after administration of brolucizumab, comprising:
Measuring the concentration of MMP-9, P-selectin, CCL7, and G-CSF in the serum isolated from the subject before administration of brolucizumab, and the percentage of Th2 cells in the blood CD4-positive lymphocytes;
A step of estimating the possibility of developing endophthalmitis based on the measured concentrations of MMP-9, P-selectin, CCL7, and G-CSF, and the percentage of Th2 cells in CD4-positive lymphocytes in the blood The method of any one of [a1]-[a12], comprising

[b1] A method for estimating the possibility of developing a side effect after administration of brolucizumab in a subject, wherein the concentration of at least one protein in a body fluid isolated from the subject before administration of brolucizumab is used as an indicator.
[b2] The method of [b1], wherein the side effect is a side effect associated with intraocular inflammation.
[b3] The method of [b1] or [b2], wherein the side effect is non-infectious endophthalmitis.
[b4] Any of [b1] to [b3], in which the side effect is selected from the group consisting of uveitis, retinal vasculitis, iritis, anterior chamber inflammation, vitreous inflammation, and iridocyclitis The method described in item 1.
[b5] Any one of [b1] to [b4], wherein the body fluid is selected from the group consisting of blood, plasma, serum, saliva, urine, sweat, tears, vitreous humor, anterior aqueous humor, and posterior aqueous humor The method described in the section.
[b6] of [b1] to [b5], wherein the protein comprises at least one protein selected from the group consisting of MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 A method according to any one of paragraphs.
[b7] indicative of at least one characteristic selected from the group consisting of low MCP-1 levels, high IP-10 levels, high P-selectin levels, high IL-2 levels, high TNFα levels, and low IL-5 levels The method according to any one of [b1] to [b6].
[b8] the subject has at least one of age-related macular degeneration (AMD), diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity, [b1] The method according to any one of to [b7].
[b9] The method of any one of [b1]-[b8], wherein the subject has been administered aflibercept prior to administration of brolucizumab.
[b10] A method for estimating the likelihood of developing endophthalmitis in a subject after administration of brolucizumab, comprising:
Low MCP-1 concentration, high IP-10 concentration, high P-selectin concentration, high IL-2 concentration, high TNFα concentration, and low IL-5 concentration in the anterior aqueous humor before brolucizumab administration,
Subject has at least one of age-related macular degeneration (AMD), diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity, [b1]-[b9 ] The method according to any one of the above.
[b11] The following formula:
a×log ₁₀ (MCP-1)＋b×log ₁₀ (IP-10)＋c×log ₁₀ (P-selectin)＋d×log ₁₀ (IL-2)＋e×log ₁₀ (TNFα)＋f×log ₁₀ (IL- 5) + g > 0
(Wherein, MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 each represent the protein concentration (pg/mL) in the body fluid, a = -17 to -21, b = 9.0 to 11.0, c = 6.4 to 7.9, d = 6.7 to 8.1, e = 6.2 to 7.6, f = -3.9 to -4.7, and g = 14 to 18).
The method according to any one of [b1] to [b10], which estimates the possibility of developing a side effect due to
[b12] The following formula:
-19 x log ₁₀ (MCP-1) + 10 x log ₁₀ (IP-10) + 7.2 x log ₁₀ (P-selectin) + 7.4 x log ₁₀ (IL-2) + 6.9 x log ₁₀ (TNFα) -4.3 x _log10 (IL-5) + 16 > 0
(wherein MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 each represent the protein concentration in the anterior aqueous humor (pg/mL))
The method according to any one of [b1] to [b11], which estimates the possibility of developing endophthalmitis due to
[b13] A method of estimating the likelihood of developing endophthalmitis in a subject after administration of brolucizumab, comprising:
measuring the concentration of MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 in pre-brolucizumab anterior aqueous humor isolated from a subject;
Based on the measured concentrations of MCP-1, IP-10, P-selectin, IL-2, TNFα and IL-5, [b1] to [ b12].
[b14] A kit for use in the method according to any one of [b1] to [b13] above,
A kit comprising a capture antibody for an indicator protein immobilized on a solid phase support, and a detection antibody for the indicator protein.
[b15] The kit of [b14], wherein the indicator proteins are MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5.
[b16] A device for estimating the likelihood of developing a side effect in a subject after administration of brolucizumab, comprising:
an information acquisition unit that acquires information on at least one protein concentration in a body fluid before administration of brolucizumab;
An apparatus comprising an estimation unit for estimating the possibility of developing a side effect based on protein concentration information, and a notification unit for outputting the estimation result.
[b17] is estimated by the following formula:
-19 x log ₁₀ (MCP-1) + 10 x log ₁₀ (IP-10) + 7.2 x log ₁₀ (P-selectin) + 7.4 x log ₁₀ (IL-2) + 6.9 x log ₁₀ (TNFα) -4.3 x _log10 (IL-5) + 16 > 0
(wherein MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 each represent the protein concentration in the anterior aqueous humor (pg/mL))
The device according to [b16], performed based on.
[b18] A program for estimating the likelihood of developing a side effect after administration of brolucizumab in a subject, comprising:
obtaining information on at least one protein concentration in the bodily fluid prior to administration of brolucizumab;
Based on the protein concentration information, the step of estimating the possibility of developing side effects,
A program that executes a process of outputting an estimation result.
[b19] is estimated by the following formula:
-19 x log ₁₀ (MCP-1) + 10 x log ₁₀ (IP-10) + 7.2 x log ₁₀ (P-selectin) + 7.4 x log ₁₀ (IL-2) + 6.9 x log ₁₀ (TNFα) -4.3 x _log10 (IL-5) + 16 > 0
(wherein MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 each represent the protein concentration in the anterior aqueous humor (pg/mL))
The program according to [b18], which is carried out based on
[b20] A method for assisting in determining whether or not to administer brolucizumab to a subject, wherein the concentration of at least one protein in a body fluid isolated from the subject prior to administration of brolucizumab is used as an index.
[b21] A method for supporting determination of the need for concomitant use of an anti-inflammatory agent when administering brolucizumab to a subject, wherein the concentration of at least one protein in a body fluid isolated from the subject prior to administration of brolucizumab is used as an index Method.
[b22] A method for estimating the possibility of developing a side effect after administration of brolucizumab in a subject, wherein the Th1/Th2 ratio in blood isolated from the subject before administration of brolucizumab is used as an index.

FIG. 1A shows IOI and non-IOI eyes after switching from administration of aflibercept to administration of brolucizumab in patients with neovascular age-related macular degeneration, and control eyes, as described in Example 1. 1 is a graph showing the concentrations of individual proteins in the anterior aqueous humor in . A: MCP-1, B: MCP-3, C: CXCL1, D: IP-10, E: CXCL13, F: G-CSF, G: GM-CSF, H: IFN-γ, I: IL-1α, J: IL-1β, K: IL-2, L: IL-5, M: IL-6, N: IL-8, O: IL-10, P: IL-12 p70, Q: IL-17A, R : MMP-1, S: MMP-9, T: ICAM-1, U: E-selectin, V: P-selectin, W: TNF-α, and X: VEGF-A. FIG. 1B shows IOI and non-IOI eyes after switching from administration of aflibercept to administration of brolucizumab in patients with neovascular age-related macular degeneration, and control eyes, as described in Example 2. 1 is a graph showing the concentrations of individual proteins in the anterior aqueous humor in . A: MCP-1, B: MCP-3, C: CXCL1, D: IP-10, E: CXCL13, F: G-CSF, G: GM-CSF, H: IFN-γ, I: IL-1α, J: IL-1β, K: IL-2, L: IL-5, M: IL-6, N: IL-8, O: IL-10, P: IL-12 p70, Q: IL-17A, R : MMP-1, S: MMP-9, T: ICAM-1, U: E-selectin, V: P-selectin, W: TNF-α, and X: VEGF-A. FIG. 2 depicts IOI and non-IOI after switching from administration of aflibercept to administration of brolucizumab in patients with neovascular age-related macular degeneration, as described in Example 2, at any time after switching. is a graph showing the concentrations of individual proteins in the serum of . A: MCP-1, B: MCP-3, C: CXCL1, D: IP-10, E: CXCL13, F: G-CSF, G: GM-CSF, H: IFN-γ, I: IL-1α, J: IL-1β, K: IL-2, L: IL-5, M: IL-6, N: IL-8, O: IL-10, P: IL-12 p70, Q: IL-17A, R : MMP-1, S: MMP-9, T: ICAM-1, U: E-selectin, V: P-selectin, W: TNF-α, and X: VEGF-A. FIG. 3 depicts the post-switch arbitrary timing of IOI and non-IOI after switching from administration of aflibercept to administration of brolucizumab in patients with neovascular age-related macular degeneration, as described in Example 2. It is a graph which shows the blood CD4-positive lymphocyte fraction. A: Th1: IFNγ+/IL4−, B: Th2: IFNγ−/IL4+, C: Th0: IFNγ+/IL4+, D: IFNγ−/IL4−, and E: Th1/Th2 ratio. The vertical axis represents the ratio (%) to the blood CD4-positive lymphocyte count. Only the Th1/Th2 ratio is unitless. 4A is a graph showing the area under the receiver operating characteristic (ROC) curve (AUC) for IOI estimation in Example 1. FIG. 4B is a graph showing the area under the receiver operating characteristic (ROC) curve (AUC) for estimation of IOI by anterior aqueous humor in Example 2. FIG. 5 is a graph showing the receiver operating characteristic (ROC) area under the curve (AUC) for estimation of IOI by serum cytokines in Example 2. FIG. 6A is a graph showing the area under the receiver operating characteristic (ROC) curve (AUC) for estimation of IOI by blood CD4-positive lymphocyte fraction in Example 2. FIG. 6B is a graph showing the receiver operating characteristic (ROC) area under the curve (AUC) for estimation of IOI by combination of serum cytokines and blood CD4-positive lymphocyte fraction in Example 2. FIG. FIG. 7 is a diagram showing one aspect of an estimation device. FIG. 8 is a diagram illustrating an example of a hardware configuration that can implement processing according to the present disclosure. FIG. 9 is a diagram showing an example of estimation processing by a computer.

The following describes in detail the modes for implementing the estimation method according to the present application, the kit used therefor, the estimation device, the estimation program, etc., with reference to the drawings. Note that the estimation method, estimation device, estimation program, etc. of the present invention are not limited by the aspects of the present disclosure.

(Method for estimating onset of side effects)
An estimation method according to one aspect of the present disclosure is a method of estimating the possibility of developing a side effect after administration of brolucizumab in a subject, wherein at least one protein concentration in a body fluid isolated from the subject before administration of brolucizumab and / Or relates to a method of using the ratio of Th2 cells in CD4-positive lymphocytes as an index.

Side effects of brolucizumab include, for example, side effects associated with intraocular inflammation, especially endophthalmitis. Endophthalmitis, a generic term for intraocular inflammation, is classified into extrinsic (injury) and endogenous (other than trauma). Endogenous endophthalmitis is further classified into infectious endophthalmitis endophthalmitis) and non-infectious endophthalmitis (synonymous with uveitis, which is gradually being called intraocular inflammation). Endophthalmitis includes, for example, retinal vasculitis, iritis, anterior chamber inflammation, vitreous inflammation, iridocyclitis, etc., which are also listed as side effects in the package insert of brolucizumab. Diagnosis of endophthalmitis can be made by confirming cells in the anterior chamber with a slit lamp microscope (if cells are confirmed, endophthalmitis is diagnosed), and inflammation of the fundus by fluorescence fundus angiography. In addition, follow-up can distinguish between infectious and non-infectious disease. For example, after exacerbation, if immunosuppression causes further exacerbation, it can be determined to be infectious.

In the estimation method according to one aspect of the present disclosure, the body fluid may be blood, plasma, serum, saliva, urine, sweat, tears, vitreous humor, anterior aqueous humor, or posterior aqueous humor. In some embodiments, aqueous humor, particularly anterior aqueous humor, may be used favorably. The area from the cornea to the iris is called the anterior chamber, and the area from the iris to the lens is called the posterior chamber, and aqueous humor is distributed in both of them. In some embodiments, multiple bodily fluid samples may be used in combination, eg, blood and serum, serum and anterior aqueous humor, anterior aqueous humor and blood, blood, serum and anterior aqueous humor. Use of such combinations may enable more accurate predictions.

As described above, the present inventors investigated age-related macular degeneration (AMD) patients who received brolucizumab injections, and examined eyes that developed endophthalmitis after brolucizumab injection and those that developed endophthalmitis immediately before brolucizumab injection. A study of differences in anterior aqueous humor protein profiles in eyes without brolucizumab revealed that patients who developed endophthalmitis after brolucizumab injection had specific proteins in the anterior aqueous humor (MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A) found a characteristic profile in the concentration. In addition, according to the knowledge obtained by the present inventors, the incidence of endophthalmitis was 7.9%, and 3 out of 48 cases of simultaneous binocular administration had bilateral onset and no unilateral onset. Assuming that the disease develops either at the same time or at the same time, the hypothesis that the disease develops separately in both eyes is rejected with p = 0.000012, so it can be inferred that the inflammation is not specific to the eye, but is due to the constitution of the eye. As a result of post hoc blood sampling to examine differences in serum protein profiles, specific proteins (IL-6, MMP-9, and IL-1β ) has a characteristic profile. As a result of examining the fraction of CD4-positive cells, we found that patients who developed endophthalmitis after brolucizumab injection had a significantly lower proportion of Th2 cells in their blood. Since it is thought that there is a constitutional predisposition, as a prediction methodology, if the profile found in post-hoc blood sampling is observed before administration, it will be the basis for discontinuing brolucizumab.

In other words, the estimation method according to the present disclosure can also be said to be an in vitro examination method for the possibility of developing a side effect, such as endophthalmitis, in a subject after administration of brolucizumab. The method according to the present disclosure can also be said to be a method of obtaining an index of the possibility of developing side effects after administration of brolucizumab in a subject, or a method of testing the likelihood of developing side effects after administration of brolucizumab to a subject.

Measurement of protein concentration in body fluids isolated from a subject, such as anterior aqueous humor, serum, or blood, can be performed by any method known to those skilled in the art, such as the ELISA method. And IP-10 that induces Th1 cells or is secreted from Th1 cells, P-selectin, and VEGF-A tend to be high, and MCP-1 that induces Th2 cells or is secreted from Th2 cells Since it tended to be low, Th1 and Th2 counts in blood samples, or the percentage of Th2 cells among CD4-positive lymphocytes, can also be used as indicators. Therefore, one aspect of the present disclosure is a method for estimating the possibility of developing a side effect after administration of brolucizumab in a subject, comprising: Th1, Th2 count, or CD4 in the blood before administration of brolucizumab isolated from the subject The present invention relates to a method (or device or program) that uses the ratio of Th2 cells in positive lymphocytes as an index. More specifically, a small percentage of Th2 cells among CD4-positive lymphocytes can be taken as an indicator. In addition, in such methods, a high Th1/Th2 ratio value can also indicate the possibility of developing side effects. Here, "a high Th1/Th2 ratio value" means, for example, a value higher than a preset threshold, and a person skilled in the art can select an appropriate threshold. The Th1/Th2 ratio in blood can be determined, for example, by quantifying cell numbers by known methods such as flow cytometry. Thus, the above method may comprise a step of quantifying the Th1/Th2 ratio in blood, eg, quantifying cell numbers by known methods such as flow cytometry. In fertility treatment, Th1 attacks the fetus and placenta, and the higher the Th1/Th2 ratio, the more difficult it is to maintain pregnancy. Similar pretreatment, ie, treatment with an immunosuppressive agent such as tacrolimus, may be considered in cases in which brolucizumab is ineffective and brolucizumab is critically needed. In some embodiments, the estimation methods of the present disclosure comprise at least one protein concentration in a pre-brolucizumab body fluid isolated from a subject, e.g., anterior aqueous humor, serum, or blood, and/or in CD4-positive lymphocytes measuring the percentage of Th2 cells in the cell, for example, by an ELISA method. The subject may be a mammal, particularly a human, particularly a patient with wet (neovascular) age-related macular degeneration.

In some embodiments, measuring both the protein concentration and the percentage of Th2 cells among CD4-positive lymphocytes may enable more accurate predictions. In this case, serum levels of MMP-9, P-selectin, CCL7, and G-CSF can be used as protein levels. More specifically, high MMP-9 concentration, high P-selectin concentration, low CCL7 concentration, and low G-CSF concentration in serum can be used as indicators.

In some embodiments of the present disclosure, the proteins used as indicators in the estimation method include at least one selected from the group consisting of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A; at least two (e.g., MCP-1 and IP-10, MCP-1 and P-selectin, MCP-1 and CXCL1, MCP-1 and VEGF-A, IP-10 and P-selectin, IP-10 and CXCL1, IP-10 and VEGF-A, P-selectin and CXCL1, P-selectin and VEGF-A, or CXCL1 and VEGF-A), at least three (e.g., MCP-1 and IP-10 and P-selectin, MCP- 1 and IP-10 and CXCL1, MCP-1 and IP-10 and VEGF-A, MCP-1 and P-selectin and CXCL1, MCP-1 and P-selectin and VEGF-A, MCP-1 and CXCL1 and VEGF- A, IP-10 and P-selectin and CXCL1, IP-10 and P-selectin and VEGF-A, IP-10 and CXCL1 and VEGF-A, or P-selectin and CXCL1 and VEGF-A), at least 4 ( For example, MCP-1 and IP-10 and P-selectin and CXCL1, MCP-1 and IP-10 and P-selectin and VEGF-A, MCP-1 and P-selectin and CXCL1 and VEGF-A, MCP-1 and IP-10 and CXCL1 and VEGF-A, or IP-10 and P-selectin and CXCL1 and VEGF-A), or all five proteins. And, in one aspect of the present disclosure, all of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A are used as indicators. These proteins are particularly suitable for use when the sample is anterior aqueous humor. Also, in some aspects of the present disclosure, in addition to some or all of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A, the concentration of other proteins may be added to the index . Other proteins that can be used as indicators include, for example, proteins showing a significant difference between samples with and without side effects, such as MMP-1, TNF-α, IL-5, ICAM-1, IL- 2, IL-1α, IFN-γ, and MCP-3, and the like, but are not limited to these. Each of these proteins may be used alone as an indicator, or may be used as an indicator in combination with other proteins (eg, MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A). Thus, in some embodiments of the present disclosure, proteins used as indicators in the estimation method are selected from the group consisting of MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5. at least 1, at least 2, at least 3, at least 4, or all 5 proteins. In one aspect of the present disclosure, all of MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 can be used as indicators. In addition, in some embodiments of the present disclosure, in addition to some or all of MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5, the concentration of other proteins is used as an indicator You can add When the sample is serum, at least one or at least two selected from the group consisting of IL-6, MMP-9, and IL-1β (for example, IL-6 and MMP-9, IL-6 and IL -1β, or MMP-9 and IL-1β), or all three proteins may suitably be used. In one aspect of the present disclosure, IL-6, MMP-9, and IL-1β are all used as indicators. A protein that can be used as an index can also be selected using a known multivariate analysis method. Specifically, variables are selected by a stepwise method, etc. (required when the number of measured protein types is larger than the number of samples in the IOI group), multivariate analysis, multiple regression analysis (when quantified using a method), it can be analyzed by methods such as discriminant analysis, logistic regression analysis, and factor analysis, and techniques such as principal component analysis, cluster analysis, and machine learning. An example of such an analysis is provided in the Examples herein by stepwise selection of variables and a nominal logistic regression analysis using a likelihood ratio test to generate an equation. The variable selected by the stepwise method does not necessarily have a low p-value, such as TNF-α shown in the examples. Such results are often obtained in multivariate analysis.

　MCP-1 is a monocyte chemoattractant and a type of chemokine. IP-10 is a chemokine that migrates activated T cells and NK cells and is involved in Th1-type inflammation. P-selectin is a protein classified as a cell surface lectin involved in adhesion between leukocytes and vascular endothelial cells, expressed in platelets and vascular endothelial cells, and involved in Th1 and Th2 type inflammation. Interleukin-2 (IL-2) is one of cytokines and is classified into a group called Th1 cytokines. TNFα is a tumor necrosis factor and a type of cytokine. Interleukin-5 (IL-5) is a type of cytokine, a Th2 cytokine that regulates humoral immunity. CXCL1 is a cytokine that causes leukocyte migration. VEGF-A is a cytokine that promotes angiogenesis.

As an indicator, in some aspects of the present disclosure, at least one selected from the group consisting of low MCP-1 concentration, high IP-10 concentration, high P-selectin concentration, high CXCL1 concentration, and high VEGF-A concentration , at least two (e.g., MCP-1 and IP-10, MCP-1 and P-selectin, MCP-1 and CXCL1, MCP-1 and VEGF-A, IP-10 and P-selectin, IP-10 and CXCL1 , IP-10 and VEGF-A, P-selectin and CXCL1, P-selectin and VEGF-A, or CXCL1 and VEGF-A), at least three (e.g., MCP-1 and IP-10 and P-selectin, MCP -1 and IP-10 and CXCL1, MCP-1 and IP-10 and VEGF-A, MCP-1 and P-selectin and CXCL1, MCP-1 and P-selectin and VEGF-A, MCP-1 and CXCL1 and VEGF -A, IP-10 and P-selectin and CXCL1, IP-10 and P-selectin and VEGF-A, IP-10 and CXCL1 and VEGF-A, or P-selectin and CXCL1 and VEGF-A), at least 4 (For example, MCP-1 and IP-10 and P-selectin and CXCL1, MCP-1 and IP-10 and P-selectin and VEGF-A, MCP-1 and P-selectin and CXCL1 and VEGF-A, MCP-1 and IP-10 and CXCL1 and VEGF-A, or IP-10 and P-selectin and CXCL1 and VEGF-A), or all five features are used. These protein concentrations are particularly suitable for use when the sample is anterior aqueous humor. And, in one aspect of the present disclosure, low MCP-1 concentration, high IP-10 concentration, high P-selectin concentration, high CXCL1 concentration, and high VEGF-A concentration are all used as indicators. Here, "low density" means, for example, a density lower than a preset threshold. Examples of thresholds are the average value of non-IOI, the threshold that gives the point with the smallest distance from the upper left point of the ROC curve (the point with sensitivity 1 and specificity 1), and the point that gives the maximum Youden Index. Examples include a threshold and a threshold at which at least one of sensitivity and specificity is 80% or higher. Similarly, "high concentration" means a concentration higher than a preset threshold value, and a person skilled in the art can select an appropriate threshold value. Whether the protein concentration is high or low can be determined, for example, based on the measured value in a control, and such a reference value can vary depending on the characteristics (race, sex, etc.) of the target population. It should be noted that, in some embodiments of the present disclosure, in addition to some or all of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A, even if the level of other protein concentrations is added to the index good. Thus, as an indication, in some aspects of the present disclosure, low MCP-1 levels, high IP-10 levels, high P-selectin levels, high IL-2 levels, high TNFα levels, and low IL-5 levels. At least one, at least two, at least three, at least four, at least five, or all six features selected from the group may be used. And, in one aspect of the present disclosure, low MCP-1 concentration, high IP-10 concentration, high P-selectin concentration, high IL-2 concentration, high TNFα concentration, and low IL-5 concentration are all used as indicators sell. When the sample is serum, at least one or at least two selected from the group consisting of low IL-6 concentration, high MMP-9 concentration, and high IL-1β concentration (for example, IL-6 and MMP- 9, IL-6 and IL-1β, or MMP-9 and IL-1β), or features of all three may suitably be used.

In some aspects of the disclosure, the subject has at least one of age-related macular degeneration (AMD), diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity. ing. Age-related macular degeneration (AMD) can specifically be wet AMD. Post hoc analyzes of data from the HAWK and HARRIER trials show that brolucizumab is also effective in patients with PCV. However, when we injected patients with brolucizumab, we saw the development of sterile endophthalmitis in 26 patients (29 eyes). The estimation method according to the present disclosure will be very useful in finding patients at risk of such endophthalmitis.

In some aspects of the present disclosure, the subject has been administered aflibercept prior to administration of brolucizumab. Aflibercept, a recombinant fusion protein consisting of the extracellular domains of the

human VEGF receptors

1 and 2 proteins and the Fc portion of the human antibody IgG1, inhibits vascular endothelial growth factor (VEGF) and thus exudative growth factor. Used in the treatment of age macular degeneration. On the other hand, brolucizumab, which has the structure of a single-chain antibody fragment (scFv), as described above, has a smaller molecular weight and is more soluble, allowing a greater number of drug molecules to be delivered to the eye with a similar intravitreal injection volume. Since it can be administered within the body, it is expected that the administration interval can be extended compared to conventional drugs, and the burden of hospital visits and treatment can be reduced. Therefore, switching from aflibercept to brolucizumab may have significant benefits. The estimation method according to the present disclosure will be very useful in evaluating the risks associated with such switching.

Some aspects of the present disclosure are more specifically a method of estimating the likelihood of developing a side effect, such as endophthalmitis, in a subject after administration of brolucizumab, comprising: Low MCP-1, high IP-10, high P-selectin, high CXCL1, and high VEGF-A concentrations in anterior aqueous humor and/or low IL-6, high MMP-9 concentrations in serum , and high IL-1β concentration, with at least one of age-related macular degeneration (AMD), diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity suffering from, how to Measurement of protein concentrations in both anterior aqueous humor and serum would allow more accurate predictions. In some aspects of the present disclosure, the subject may or may not have been administered aflibercept prior to administration of brolucizumab and may be presumed to develop an IOI at the first dose of brolucizumab. Some aspects of the present disclosure are a method of estimating the likelihood of developing a side effect, e.g., endophthalmitis, in a subject after administration of brolucizumab, optionally comprising: Low MCP-1 levels, high IP-10 levels, high P-selectin levels, high IL-2 levels, high TNFα levels, and low IL-5 levels in patients with age-related macular degeneration (AMD) and diabetes The method also relates to having at least one of macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity.

We also determined from analysis of proteins in the anterior aqueous humor of patients that the following formula:
－5.8×log10(MCP-1)＋2.8×log10(IP-10)＋2.6×log10(P-selectin)＋2.0×log10(CXCL1)＋1.1×log10(VEGF-A)－0.12 ＞ 0
(wherein MCP-1, IP-10, P-selectin, CXCL1 and VEGF-A each represent the protein concentration in the anterior aqueous humor (pg/mL))
It was found that the possibility of developing endophthalmitis can be estimated by If the value obtained by substituting each protein concentration into the above formula becomes a positive number, it is presumed that endophthalmitis may develop. For example, if the concentrations of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A are 980, 180, 390, 73, and 5.5, respectively, the value of the above formula is 0.12, and endophthalmitis is presumed to have the potential to develop

Also, optionally the following formulas:
-19 x log ₁₀ (MCP-1) + 10 x log ₁₀ (IP-10) + 7.2 x log ₁₀ (P-selectin) + 7.4 x log ₁₀ (IL-2) + 6.9 x log ₁₀ (TNFα) -4.3 x _log10 (IL-5) + 16 > 0
(wherein MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 each represent the protein concentration in the anterior aqueous humor (pg/mL))
may be used. According to this formula, for example, if the concentrations of MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 are 980, 180, 390, 1.9, 1.8, and 3.3, respectively, The value of the above formula is 2.0, and it is estimated that endophthalmitis may develop.

The above formula may be used after being adjusted according to the type of side effect, the type of body fluid, the characteristics of the target population (race, gender, etc.), etc. For example, the following formula:
a×log ₁₀ (MCP-1)＋b×log ₁₀ (IP-10)＋c×log ₁₀ (P-selectin)＋d×log ₁₀ (CXCL1)＋e×log ₁₀ (VEGF-A)＋f＞0
where a to f are a = -11 to -0.91, b = 0.50 to 5.2, c = 0.66 to 4.6, d = -0.17 to 4.2, e = -0.071 to 2.3, and f = Any value in the range of -12 to 11 may be used. Also, based on standard error ranges, a = -11 to -0.91, b = 0.50 to 5.2, c = 0.66 to 4.6, d = -0.17 to 4.2, e = -0.071 to 2.3, and Any value in the range of f=-12 to 11 may be used.

Also, the formula below:
a×log ₁₀ (MCP-1)＋b×log ₁₀ (IP-10)＋c×log ₁₀ (P-selectin)＋d×log ₁₀ (IL-2)＋e×log ₁₀ (TNFα)＋f×log ₁₀ (IL- 5) + g > 0
, a to g are a = -17 to -21, b = 9.0 to 11.0, c = 6.4 to 7.9, d = 6.7 to 8.1, e = 6.2 to 7.6, f = -3.9 to −4.7, and any value in the range of g=14-18. Also, based on the range of standard errors, a to e are respectively a = -12 to -26, b = 5.7 to 14, c = 3.5 to 11, d = 4.1 to 11, e = 3.5 to 10, f = - Any value in the range of 2.0 to -6.7 and g=3.7 to 28 may be used.

Especially when the body fluid is serum, the following formula can be used:
a × _log10 (IL-6) + b × _log10 (MMP-9) + c × _log10 (IL-1β) + d > 0
(In the formula, IL-6, MMP-9, and IL-1β each represent the serum protein concentration (pg/mL), a = -35 to -12, b = 1.7 to 4.7, c = 7.1 to 30, and d = -17 to 1.2).

Here, more specifically, the following formula can be used:
-23 x _log10 (IL-6) + 3.2 x _log10 (MMP-9) +19 x _log10 (IL-1β)) -8.1 > 0
(where IL-6, MMP-9, and IL-1β each represent the protein concentration in serum (pg/mL)).

Especially when the body fluid is blood, the following formula can be used:
Blood Th2 < a, where a = 2.5-4.6.

Here, more specifically, the following formula can be used:
Blood Th2 < 3.4.

In some embodiments, the following formula combining protein concentration and percentage of Th2 cells among CD4 positive lymphocytes may be used:
a × Th2 + b × log ₁₀ (MMP-9) + c × log ₁₀ (P-selectin) + d × log ₁₀ (CCL7) + e × log ₁₀ (G-CSF) + f > 0
(where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, MMP-9, P-selectin, CCL7, and G-CSF each represent protein concentration in serum (pg/mL), a = -1.6 to -0.78, b = 4.2 to 9.6, c = 12 to 31, d = -76 to -27, e = -19 to -5.3, and f = -12 to 40)

Here, more specifically, the following formula can be used:
-1.2 x Th2 + 6.9 x log ₁₀ (MMP-9) + 22 x log ₁₀ (P-selectin) -52 x log ₁₀ (CCL7) -12 x log ₁₀ (G-CSF) + 14 > 0
(Where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, and MMP-9, P-selectin, CCL7, and G-CSF are protein concentrations in serum (pg/mL), respectively.)

Some aspects of the present disclosure are a method of estimating the likelihood of developing a side effect, e.g., endophthalmitis, in a subject after administration of brolucizumab, comprising: measuring the concentration of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A in the anterior aqueous humor or, for example, the concentration of IL-6, MMP-9, and IL-1β in serum; Based on the measured levels of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A, or levels of IL-6, MMP-9, and IL-1β, side effects such as endophthalmitis It relates to a method comprising the step of estimating the likelihood of developing the disease. Concentrations of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A in body fluids, e.g., anterior aqueous humor, and/or concentrations of IL-6, MMP-9, and IL-1β in serum can be measured by any method known to those skilled in the art, such as the ELISA method. Measurement of protein concentrations in both anterior aqueous humor and serum would allow more accurate predictions. Estimation of the likelihood of developing side effects, such as endophthalmitis, can be made according to the formulas disclosed herein, based on protein concentration measurements. Note that in some aspects of the present disclosure, instead of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A, MCP-1, IP-10, P-selectin, IL-2, TNFα , and concentrations of IL-5 can be used.

If it is presumed that side effects such as endophthalmitis may develop in the subject after administration of brolucizumab, for example, it may be decided not to administer brolucizumab to the subject. In this case, the subject may be treated with an agent other than brolucizumab, such as aflibercept. In addition, if the subject is not expected to develop side effects such as endophthalmitis after administration of brolucizumab, for example, brolucizumab may be administered to the subject. In the case of endophthalmitis, steroids may be used in combination if the effect of other drugs is insufficient. In addition, when endophthalmitis develops in a subject, treatment with, for example, steroids can be performed.

(Kit for use in estimating the onset of endophthalmitis)
Some aspects of the present disclosure are a method of estimating the likelihood of developing a side effect, e.g., endophthalmitis, in a subject after administration of brolucizumab, comprising: It relates to a kit for use in a method in which the concentration of at least one protein in the anterior aqueous humor, serum, or blood and/or the percentage of Th2 cells in CD4-positive lymphocytes is used as an index. A kit according to the present disclosure includes, for example, an indicator protein capture antibody immobilized on a solid support and an indicator protein detection antibody. By using the kit according to the present disclosure, the concentration of the indicator protein can be measured.

In some embodiments of the present disclosure, the indicator protein includes at least one, at least two selected from the group consisting of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A (e.g. , MCP-1 and IP-10, MCP-1 and P-selectin, MCP-1 and CXCL1, MCP-1 and VEGF-A, IP-10 and P-selectin, IP-10 and CXCL1, IP-10 and VEGF -A, P-selectin and CXCL1, P-selectin and VEGF-A, or CXCL1 and VEGF-A), at least three (e.g., MCP-1 and IP-10 and P-selectin, MCP-1 and IP-10 and CXCL1, MCP-1 and IP-10 and VEGF-A, MCP-1 and P-selectin and CXCL1, MCP-1 and P-selectin and VEGF-A, MCP-1 and CXCL1 and VEGF-A, IP-10 and P-selectin and CXCL1, IP-10 and P-selectin and VEGF-A, IP-10 and CXCL1 and VEGF-A, or P-selectin and CXCL1 and VEGF-A), at least four (e.g., MCP-1 and IP-10 and P-selectin and CXCL1, MCP-1 and IP-10 and P-selectin and VEGF-A, MCP-1 and P-selectin and CXCL1 and VEGF-A, MCP-1 and IP-10 and CXCL1 and VEGF-A, or IP-10 and P-selectin and CXCL1 and VEGF-A), or all five. These proteins are particularly suitable for use when the sample is anterior aqueous humor. Thus, in some embodiments, kits of the present disclosure include at least one, at least two, at least Included are solid supports having immobilized antibodies to capture three, at least four, or all five proteins, and antibodies to detect the captured proteins. Solid phase supports can be, for example, plates, beads, membranes, etc., but are not limited to these. The kit according to the present disclosure can be used to measure the concentration of an index protein, and based on the value, the possibility of developing endophthalmitis can be estimated. Such kits can also be used as companion diagnostics. Note that in some aspects of the present disclosure, instead of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A, MCP-1, IP-10, P-selectin, IL-2, TNFα , and IL-5 can be used as indicators. When the sample is serum, at least one or at least two selected from the group consisting of IL-6, MMP-9, and IL-1β (for example, IL-6 and MMP-9, IL-6 and IL -1β, or MMP-9 and IL-1β), or all three proteins may suitably be used. In one aspect of the present disclosure, IL-6, MMP-9, and IL-1β are all used as indicators.

(Apparatus for estimating the possibility of developing endophthalmitis)
Some aspects of the present disclosure are a device for estimating the likelihood of developing a side effect, e.g., endophthalmitis, in a subject after administration of brolucizumab, comprising: or at least one protein concentration in blood and/or an information acquisition unit for obtaining information on the percentage of Th2 cells in CD4-positive lymphocytes, based on the information on the protein concentration and/or the percentage of Th2 cells in CD4-positive lymphocytes , an estimation unit for estimating the possibility of developing a side effect, for example, endophthalmitis, and a notification unit for outputting the estimation result.

The device 100 illustrated in FIG. 7 has an information acquisition unit 10, an estimation unit 20, and a notification unit 30. The device 100 can have an arithmetic device such as a CPU (Central Processing Unit) and a storage device such as a hard disk and flash memory. This storage device can store a program for controlling the device and acquired data (protein concentration data, etc.). The device 100 may work in cooperation with the computing and storage devices of a server connected by a network. All elements of the apparatus (system) according to the present disclosure do not need to be physically located at the same location, and may be connected via a network.

The information acquisition unit 10 illustrated in FIG. 7 obtains information on the concentration of at least one protein in body fluids, such as anterior aqueous humor, serum, or blood, and/or the ratio of Th2 cells in CD4-positive lymphocytes before administration of brolucizumab. can be obtained. More specifically, the information is obtained by electronically importing the data of measurements such as, for example, ELISA, or by manual entry from a keyboard.

The estimation unit 20 exemplified in FIG. 7 can estimate the possibility of developing side effects such as endophthalmitis based on information on the protein concentration and/or the ratio of Th2 cells in CD4-positive lymphocytes. The estimation can be performed, for example, based on the level of the protein concentration used as an index. In more detail, for example, the following formula: a × log ₁₀ (MCP-1) + b × log ₁₀ (IP-10) + c × log ₁₀ (P-selectin) + d × log ₁₀ (CXCL1) + e × log ₁₀ (VEGF-A) + f > 0 (wherein MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A are each in body fluids, for example, the protein concentration in the anterior aqueous humor (pg/mL) a = -11 to -0.91, b = 0.50 to 5.2, c = 0.66 to 4.6, d = -0.17 to 4.2, e = -0.071 to 2.3, and f = -12 to 11). Estimate the possibility of developing side effects, for example, endophthalmitis. Also, a to e are a = -11 to -0.91, b = 0.50 to 5.2, c = 0.66 to 4.6, d = -0.17 to 4.2, e = -0.071 to 2.3, and f = -12 to 11 respectively. Any value in the range is acceptable. More specifically, for example, the following formula: −5.8×log10(MCP-1)+2.8×log10(IP-10)+2.6×log10(P-selectin)+2.0×log10(CXCL1)+1. 1 × log10 (VEGF-A) - 0.12 > 0 (where MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A each represent the protein concentration in the anterior aqueous humor (pg/mL) ) can estimate the possibility of developing endophthalmitis.

In some cases, the following formula: a × log ₁₀ (MCP-1) + b × log ₁₀ (IP-10) + c × log ₁₀ (P-selectin) + d × log ₁₀ (IL-2) + e × log ₁₀ ( TNFα) + f × log ₁₀ (IL-5) + g > 0, where MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 are each in body fluids, e.g. Expresses protein concentration in water (pg/mL); 6.7, and g=3.7-28). a to e are respectively a = -17 to -21, b = 9.0 to 11.0, c = 6.4 to 7.9, d = 6.7 to 8.1, e = 6.2 to 7.6, f = -3.9 to -4.7, and g = 14 It can be any value in the range of ~18. More specifically, for example, the following formula: -19 x log10 (MCP-1) + ₁₀ x log10 (IP- ₁₀ ) + 7.2 x _log10 (P-selectin) + 7.4 x _log10 (IL- 2) + 6.9 x log ₁₀ (TNFα) - 4.3 x log ₁₀ (IL-5) + 16 > 0, where MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 represents the protein concentration (pg/mL) in the anterior aqueous humor), the possibility of developing endophthalmitis may be estimated.

Especially when the body fluid is blood, the following formula can be used:
Blood Th2 < a (where Th2 represents the percentage of Th2 cells in blood CD4-positive lymphocytes, a=2.5-4.6).

Here, more specifically, the following formula can be used:
Blood Th2 < 3.4 (where Th2 represents the percentage of Th2 cells in blood CD4-positive lymphocytes).

The notification unit 30 illustrated in FIG. 7 can output the estimation result. The notification unit 30 may include a display, printer, speaker, etc. for presenting information to the user. Also, the output destination of the notification unit 30 may be another server or device connected via a network. For example, the output from the notification unit 30 may be sent directly to an electronic medical record system or the like.

Here, in the above-described apparatus, an execution program that allows a computer to execute each function is generated, and the execution program is installed in, for example, a general-purpose personal computer, a server, etc., or a combination thereof, thereby performing processing. can do.

A general-purpose personal computer, for example, has an IO device (input device, output device), a storage device (drive device, auxiliary storage device), a memory device (RAM, ROM), a CPU (Central Processing Unit) that performs various controls, It is configured with network connection devices, which can be interconnected with a system bus (Fig. 8). The input device has a pointing device such as a keyboard and a mouse operated by a user or the like, and inputs various operation signals such as execution of a program from the user or the like. The input device can also input various data such as protein concentration data obtained via a communication network from an external device connected to a network connection device or the like. The output device has a display that displays various windows and data necessary for operating the computer main body for performing the processing related to the present disclosure, and displays the execution progress and results of the program by the control program held by the CPU. can do. Further, the output device can print the above-described processing results and the like on a print medium such as paper and present them to the user and the like.

Here, the execution program installed in the computer main body in the present disclosure is provided by recording media such as USB (Universal Serial Bus) memory, CD-ROM, DVD, etc., for example. A recording medium recording a program can be set in a drive device, and an execution program contained in the recording medium is installed in an auxiliary storage device from the recording medium via the drive device. Also, the execution program may be downloaded in whole or in part from an external device via any network when necessary. Furthermore, the external device may be provided with an auxiliary storage device in which the execution program is installed and/or a memory device that stores the installed execution program. The external device may be a server. The server may be a specific server, or may be an indefinite server such as a cloud infrastructure. Furthermore, the execution program may be provided by a subscription type service. A subscription-type service is one of the usage forms of computer software, and is a service in which fees are paid according to the period of use of the software. In this way, the execution program may be used in any form and is not limited.

The auxiliary storage device is storage means such as a hard disk, and can store the execution program according to the present disclosure, the control program provided in the computer, etc., and can input/output as necessary. The memory device stores an execution program and the like read from the auxiliary storage device by the CPU. The memory device is composed of ROM (Read Only Memory), RAM (Random Access Memory), and the like. Based on control programs such as the OS (Operating System) and execution programs stored in the memory device, the CPU controls the processing of the entire computer, such as various calculations and data input/output with each hardware component. Each process can be realized by Various information required during program execution can be acquired from an auxiliary storage device, and execution results can be stored. By connecting to a communication network or the like, the network connection device acquires an execution program from another terminal or the like connected to the communication network, or obtains an execution result obtained by executing the program or the execution according to the present disclosure. The program itself can be provided to another terminal or the like.

With the hardware configuration as described above, the estimation processing according to the present disclosure can be executed. Also, by installing the program, the processing according to the present disclosure can be easily implemented on a general-purpose personal computer or the like.

(Program to estimate the possibility of developing endophthalmitis)
Some aspects of the present disclosure are a program for estimating the likelihood of developing a side effect, e.g., endophthalmitis, in a subject after administration of brolucizumab, wherein a computer stores obtaining information of at least one protein concentration and/or percentage of Th2 cells among CD4-positive lymphocytes in serum or blood; The present invention relates to a program for executing a step of estimating the possibility of developing endophthalmitis and a step of outputting the estimation result. The program according to the present disclosure may be recorded on a computer readable medium having non-transitory instructions stored thereon that, when executed by a processor, perform the steps described above. FIG. 9 shows an example of program processing, and steps 100 (S100) to step 120 (S120) are processed by the CPU of a computer (which may include a server) executing the program. Some aspects of the present disclosure are a computer-implemented method for estimating the likelihood of developing a side effect, e.g., endophthalmitis, in a subject after administration of brolucizumab, comprising: For example, obtaining information on at least one protein concentration in anterior aqueous humor, serum, or blood and/or the percentage of Th2 cells in CD4-positive lymphocytes, protein concentration and/or Th2 cells in CD4-positive lymphocytes It can also be expressed as a method including a step of estimating the possibility of developing endophthalmitis based on information on the ratio of , and a step of outputting the estimation result.

The estimation process exemplified in FIG. 9 obtains information on the concentration of at least one protein and/or the percentage of Th2 cells in CD4-positive lymphocytes in body fluids, e.g., anterior aqueous humor, serum, or blood, before administration of brolucizumab. Step (S100), based on information on the protein concentration and / or the ratio of Th2 cells in the CD4 positive lymphocytes, the step of estimating the possibility of developing side effects, for example, endophthalmitis (S110), and the estimation result It includes an output step (S120).

Obtaining information on the concentration of at least one protein in body fluids before administration of brolucizumab illustrated in FIG. ), the computer obtains information on the concentration of at least one protein and/or the percentage of Th2 cells in CD4-positive lymphocytes in body fluids, e.g., anterior aqueous humor, serum, or blood, prior to administration of brolucizumab. Executed according to the program. More specifically, the information is obtained, for example, by performing a process of electronically importing measurement data such as ELISA, or by accepting manual input from a keyboard.

Based on the information on the protein concentration and/or the ratio of Th2 cells in the CD4-positive lymphocytes illustrated in FIG. Based on the information on the concentration and/or the ratio of Th2 cells in the CD4-positive lymphocytes, processing for estimating the possibility of developing side effects such as endophthalmitis is executed based on the program. The estimation can be performed, for example, based on the level of the protein concentration used as an index. More specifically, for example, the following formula: a × log ₁₀ (MCP-1) + b × log ₁₀ (IP-10) + c × log ₁₀ (P-selectin) + d × log ₁₀ (CXCL1) + e × log ₁₀ ( VEGF-A) + f > 0, where MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A each represent the protein concentration (pg/mL) in body fluids, e.g., anterior aqueous humor. , a = -11 to -0.91, b = 0.50 to 5.2, c = 0.66 to 4.6, d = -0.17 to 4.2, e = -0.071 to 2.3, and f = -12 to 11). For example, the possibility of developing endophthalmitis is estimated. Also, a to f are a = -11 to -0.91, b = 0.50 to 5.2, c = 0.66 to 4.6, d = -0.17 to 4.2, e = -0.071 to 2.3, and f = -12 to 11 respectively. Any value in the range is acceptable. More specifically, for example, the following formula: −5.8×log ₁₀ (MCP-1)+2.8×log ₁₀ (IP-10)+2.6×log ₁₀ (P-selectin)+2.0×log ₁₀ ( CXCL1) + 1.1 × log ₁₀ (VEGF-A) - 0.12 > 0 (where MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A are each the protein concentration in the anterior aqueous humor (pg /mL)) can be used to estimate the possibility of developing endophthalmitis.

In addition, the following formula: a × log ₁₀ (MCP-1) + b × log ₁₀ (IP-10) + c × log ₁₀ (P-selectin) + d × log ₁₀ (IL-2) + e × log ₁₀ (TNFα) + f × log ₁₀ (IL-5) + g > 0, where MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 are proteins in body fluids, e.g., anterior aqueous humor, respectively. Represents concentration (pg/mL), a = -17 to -21, b = 9.0 to 11.0, c = 6.4 to 7.9, d = 6.7 to 8.1, e = 6.2 to 7.6, f = -3.9 to -4.7, and g = 14 to 18) may be used, and a to e are respectively a = -12 to -26, b = 5.7 to 14, c = 3.5 to 11, d = 4.1 to 11, e = 3.5. ~10, f = -2.0 to -6.7, and g = 3.7 to 28. More specifically, for example, the following formula: -19 x log10 (MCP-1) + ₁₀ x log10 (IP- ₁₀ ) + 7.2 x _log10 (P-selectin) + 7.4 x _log10 (IL- 2) + 6.9 x log ₁₀ (TNFα) - 4.3 x log ₁₀ (IL-5) + 16 > 0, where MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 each represent the protein concentration in the anterior aqueous humor (pg/mL)) may be used for estimation.

Here, more specifically, the following formula can be used:
Blood Th2 < 3.4.

In the step of outputting the estimation results (S120) illustrated in FIG. 9, the process of causing the computer to output the estimation results is executed based on the program. Output may be to a display, printer, speakers, etc. for presenting information to a user. Also, the output destination may be another server or device connected via a network. For example, the output from step S120 may be sent directly to an electronic medical record system or the like.

(Methods, devices, and programs for supporting decisions on whether to administer brolucizumab)
A method according to one aspect of the present disclosure is a method of assisting in determining whether to administer brolucizumab to a subject, wherein at least one protein concentration and/or CD4 in a pre-brolucizumab body fluid isolated from the subject The present invention relates to a method using the ratio of Th2 cells in positive lymphocytes as an index. In some embodiments, the assisted method of the present disclosure comprises measuring the concentration of at least one protein and/or the percentage of Th2 cells among CD4 positive lymphocytes in a pre-brolucizumab body fluid isolated from the subject. . The description of the estimation method, estimation apparatus, and estimation program in this specification can be applied to the protein used as an index and other details. The methods of the present disclosure can provide information for physicians to decide whether to administer brolucizumab to a subject. A method according to one aspect of the present disclosure also relates to an apparatus and program for assisting in determining whether to administer brolucizumab to a subject. In some embodiments, the device/program of the present disclosure can present an output such as "administration of brolucizumab is undesirable" based on the measurement results of proteins such as cytokines. In this case, the subject may be treated with an agent other than brolucizumab, such as aflibercept. In some aspects, the device/program of the present disclosure may or may not explicitly indicate the prediction result regarding the possibility of developing side effects such as endophthalmitis.

(Method, Device, Program for Supporting Judgment of Necessity of Concomitant Use of Anti-Inflammatory Agents)
A method according to one aspect of the present disclosure is a method for supporting determination of the necessity of concomitant use of an anti-inflammatory agent (such as a steroid agent) when administering brolucizumab to a subject, wherein the administration of brolucizumab isolated from the subject It relates to a method of indexing the concentration of at least one protein in a body fluid and/or the percentage of Th2 cells in CD4-positive lymphocytes. In some embodiments, the assisted method of the present disclosure comprises measuring the concentration of at least one protein and/or the percentage of Th2 cells among CD4 positive lymphocytes in a pre-brolucizumab body fluid isolated from the subject. . For the protein to be used as an index and other details, as well as the configuration of the apparatus and program, etc., the descriptions elsewhere in this specification can be applied. The method of the present disclosure can provide information for a physician to determine whether or not an anti-inflammatory agent (such as a steroid agent) is required when administering brolucizumab to a subject. A method according to an aspect of the present disclosure also relates to a device and a program for assisting in determining whether or not an anti-inflammatory agent (such as a steroid agent) is required when administering brolucizumab to a subject. In some embodiments, the device/program of the present disclosure can present an output such as "combination of brolucizumab and an anti-inflammatory agent is desirable" based on the measurement results of proteins such as cytokines. In this case, the subject may be administered brolucizumab. In some aspects, the device/program of the present disclosure may or may not explicitly indicate the prediction result regarding the possibility of developing side effects such as endophthalmitis.

As described above, the present invention has been described with specific examples, but the above specific examples are merely examples, and the present invention can be implemented with arbitrary modifications within the scope of the claims. . The various features and aspects of the present invention, referred to elsewhere above, may be applied mutatis mutandis to the rest of the description as appropriate. Thus, features specified in one aspect may be combined with features specified in other aspects as appropriate. All references cited herein, including patents, patent applications, articles, textbooks, and sequence accession numbers, and references cited therein, are hereby incorporated by reference in their entirety. . to the extent that one or more of the incorporated literature and similar materials differ from or conflict with the present application in any respect including, but not limited to, defined terms, usage of terms, illustrated techniques, etc.; The description of the present application takes precedence.

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples.

[Example 1]
1. Clinical study The study disclosed in this example was conducted from June to December 2020 at Jichi Medical University in which nine consecutive cases of age-related macular degeneration (AMD) developed endophthalmitis after switching from aflibercept to brolucizumab ( AMD) patients (9 eyes) were analyzed by matching with 52 AMD patients (52 eyes) who switched to brolucizumab. Thirty-three patients (33 eyes) without fundus disease who underwent cataract surgery were also used as controls. This study was conducted in accordance with the Declaration of Helsinki, obtained informed consent from all patients, and obtained approval from the institutional ethics committee.

Approximately 0.2 mL of anterior aqueous humor was collected from each patient immediately before switching to brolucizumab. At the start of cataract surgery, an undiluted anterior aqueous humor sample (usually approximately 0.2 mL volume) was manually aspirated with a disposable syringe and immediately sterilized with ProteoSave™ (Sumitomo Bakelite Co., Ltd., Tokyo, Japan). Transferred to tubes and stored at -80°C until needed.

A multiplex cytokine assay was used according to the manufacturer's instructions to determine the concentrations of the following proteins: MCP-1, MCP-3, CXCL1, IP-10, CXCL13, G-CSF, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-5, IL-6, IL-8, IL-10, IL-12 p70, IL-17A, MMP-1, MMP-9, ICAM-1, E - Selectins, P-selectins, TNF-α, and VEGF-A. Their detection limits are 9.9, 3.2, 5.3, 1.18, 11.5, 4.1, 4.1, 0.40, 0.9, 0.8, 1.8, 0.5, 1.7, 1.8, 1.6, 20.2, 1.8, 2.7, 13.6, 87.9, 18.8, 9.0, respectively. , 1.2, and 2.1 pg/mL. Measurements were taken twice for each sample and the average was calculated.

Routinely perform fluorescein angiography to detect type 1 and type 2 choroidal neovascularization (CNV) in wet AMD cases, except in patients contraindicated by drug hypersensitivity, liver dysfunction, or recent cerebrovascular events. and retinal angiomatous proliferation (RAP). Indocyanine green angiography was also performed along with fluorescein angiography to identify polypoidal choroidal vasculopathy (PCV). In addition, at each patient visit, swept source optical coherence tomography (OCT); Silverstone (NIKON CORPORATION, Tokyo, Japan), DRI OCT Triton (TOPCON CORPORATION, Tokyo, Japan), or spectral domain OCT; NIDEK CO.LTD., Aichi, Japan).

2. Statistical Analysis Statistical analysis was performed using JMP Pro software version 15.2.1 (SAS Institute Inc., Cary, NC). Categorical data were evaluated using Pearson's chi-square test and continuous variables were evaluated using Student's t-test after normalization between endophthalmitis development and control after brolucizumab. After comparing with and without logarithmic transformation, the one better suited to the normal distribution form was used. Because this was a hypothesis-generating study aimed at identifying a possible association between the development of endophthalmitis after brolucizumab and anterior aqueous humor proteins, no Bonferroni correction was performed. Stepwise variable selection (minimum Bayesian information criterion, variable addition) was performed to select explanatory variables for protein concentration. A nominal logistic regression analysis with likelihood ratio test was performed on the incidence of endophthalmitis after brolucizumab. We calculated the area under the receiver operating characteristic (ROC) curve (AUC) and also calculated the sensitivity and specificity of the Youden index.

3. result
Patient demographic characteristics The average age of cases with IOI after brolucizumab (IOI cases), cases without IOI after brolucizumab (non-IOI cases; also referred to as non-IOI cases), and control cases, respectively, was 74.1 years (range, 58-87 years), 75.1 years (57-91 years, vs IOI P=0.70), and 74.0 years (56-88 years, vs AMD P=0.51). 8 of 9 IOI cases (89%), 38 of 52 non-IOI cases (73.1%, P for IOI = 0.31), and 16 of 33 controls (48.5%, P for AMD vs. AMD = 0.0086) were male. The mean duration of treatment for eyes that developed IOI (IOI eyes) and eyes that did not develop IOI (non-IOI eyes) was 46.0 and 51.7 months, respectively (P=0.65). The mean number of injections in IOI and non-IOI eyes was 18.8 and 17.5, respectively (P = 0.80). The mean final injection intervals for IOI, non-IOI, and control eyes were 10 and 11 weeks, respectively (P=0.45). All IOI cases had PCV and received aflibercept prior to switching.

Protein Concentrations in IOI Patients, Non-IOI Patients, and Control Anterior Aqueous Humor Prior to Brolucizumab Injection Protein concentrations in IOI, non-IOI, and control anterior aqueous humor are shown in FIG. 1A. Although there were no significant differences in concentrations of individual cytokines in IOI patients when compared to samples taken from non-IOI patients, some of the analyzed cytokines showed numerical differences. P-selectin (616 vs 351 pg/mL, P = 0.07), MCP-1 (823 vs 1043 pg/mL, P = 0.17), MMP-1 (9.2 vs 6.3 pg/mL, P = 0.26), TNF- α (1.3 vs 0.92 pg/mL, P = 0.26), IL-5 (2.5 vs 3.3 pg/mL, P = 0.3), ICAM-1 (1924 vs 2309 pg/mL, P = 0.4), IL-2 ( 3.1 vs 2.5 pg/mL, P = 0.4), IL-1 α (1.1 vs 0.79 pg/mL, P = 0.38), IFN-γ (2.6 vs 2.0 pg/mL, P = 0.4), and MCP-3 ( 28 vs 36 pg/mL, P = 0.5) has a P value less than or equal to 0.5.

There were significant differences in the concentrations of several cytokines in AMD eyes when compared to controls. MCP-1 (1008 vs 789 pg/mL, P = 0.017*), IP-10 (44 vs 16 pg/mL, P < 0.0001*), CXCL13 (12 vs 5.3 pg/mL, P < 0.0001*), G -CSF (17 vs 10 pg/mL, P = 0.0018*), GM-CSF (4.7 vs 2.9 pg/mL, P = 0.0096*), IFN-γ (2.1 vs 1.4 pg/mL, P = 0.042*), IL-1 α (0.83 vs 0.39 pg/mL, P = 0.0076*), IL-1β (1.3 vs 1.7 pg/mL, P = 0.014*), IL-5 (3.1 vs 4.1 pg/mL, P = 0.021*) ), IL-6 (7.6 vs 27 pg/mL, P < 0.0001*), IL-8 (15 vs 10 pg/mL, P = 0.018*), IL-10 (5.0 vs 1.7 pg/mL, P < 0.0001 *), E-selectin (32 vs 25 pg/mL, P = 0.043*), TNF-α (0.97 vs 0.38 pg/mL, P = 0.0005*), and VEGF-A (3.9 vs 25 pg/mL, P <0.0001*). On the other hand, MCP-3 (35 vs 39 pg/mL, P = 0.49), CXCL1 (42 vs 45 pg/mL, P = 0.60), IL-2 (2.5 vs 2.2 pg/mL, P = 0.39), IL- 12 p70 (24 vs 27 pg/mL, P = 0.37), IL-17A (0.97 vs 0.95 pg/mL, P = 0.95), MMP-1 (6.6 vs 7.1 pg/mL, P = 0.74), MMP-9 (151 vs 217 pg/mL, P = 0.091), ICAM-1 (2248 vs 2663 pg/mL, P = 0.30), and P-selectin (382 vs 246 pg/mL, P = 0.32) were not significant .

Nominal logistic regression analysis and ROC curve MCP-1 (coefficient normalized to concentration: -3.9, P = 0.0002), IP-10 (3.4, 0.001), P-selectin (2.6, 0.002), IL-2 (2.2) , 0.002), TNFα (2.3, 0.02), and IL-5 (−1.2, 0.05) were selected by stepwise variable selection. Nominal logistic regression analysis and Youden index yielded "-19 x log10 (MCP-1) + ₁₀ x log10 (IP- ₁₀ ) + 7.2 x _log10 (P-selectin) + 7.4 x _log10 (IL-2 ) + 6.9 x log ₁₀ (TNFα) - 4.3 x log ₁₀ (IL-5) + 16 >0' was found to affect IOI after switching to brolucizumab. AUC was 0.95, sensitivity was 89%, and specificity was 94% (Fig. 4A).

Four. CONCLUSIONS: Patients who developed endophthalmitis after switching to brolucizumab had lower concentrations of MCP-1 and IL-5 in the anterior aqueous humor before switching, and higher concentrations of IP-10, P-selectin, IL-2 and TNFα. high. Eyes that develop endophthalmitis after brolucizumab are thought to have asymptomatic cell-mediated immune-mediated inflammation before the switch.

[Example 2]
1. Clinical Studies An additional study disclosed in this example investigated 14 consecutive cases of endophthalmitis after switching from aflibercept to brolucizumab and the first The subjects were 5 consecutive cases of age-related macular degeneration (AMD) patients (19 eyes) who developed endophthalmitis after receiving brolucizumab as treatment. We analyzed 14 AMD patients (77 eyes) who received brolucizumab and were matched by age, sex, and whether or not they received brolucizumab anterior vitreous injection. In addition, 24 patients (24 eyes) without fundus disease who underwent cataract surgery were used as controls. This study was conducted in accordance with the Declaration of Helsinki, obtained informed consent from all patients, and obtained approval from the institutional ethics committee.

Approximately 0.2 mL of anterior aqueous humor was collected from each patient immediately before switching to brolucizumab. At the start of cataract surgery, an undiluted anterior aqueous humor sample (usually approximately 0.2 mL volume) was manually aspirated with a disposable syringe and immediately sterilized with ProteoSave™ (Sumitomo Bakelite Co., Ltd., Tokyo, Japan). Transferred to tubes and stored at -80°C until needed. Blood and plasma were collected from 20 each of IOI and non-IOI cases matched by age, sex, and with or without brolucizumab anterior vitreous injection from outpatient AMD patients from December 2021 to February 2022, and 22 each. Example blood was collected. Serum was treated in the same manner as anterior aqueous humor, and blood was immediately submitted to SRL Co., Ltd. (Tokyo, Japan).

A multiplex cytokine assay was used according to the manufacturer's instructions to determine the concentrations of the following proteins: MCP-1, MCP-3, CXCL1, IP-10, CXCL13, G-CSF, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-5, IL-6, IL-8, IL-10, IL-12 p70, IL-17A, MMP-1, MMP-9, ICAM-1, E - Selectins, P-selectins, TNF-α, and VEGF-A. Their detection limits are 9.9, 3.2, 5.3, 1.18, 11.5, 4.1, 4.1, 0.40, 0.9, 0.8, 1.8, 0.5, 1.7, 1.8, 1.6, 20.2, 1.8, 2.7, 13.6, 87.9, 18.8, 9.0, respectively. , 1.2, and 2.1 pg/mL. Measurements were taken twice for each sample and the average was calculated. Fractionation of CD4-positive cell water was outsourced to SRL.

Fluorescein angiography was routinely performed to distinguish between type 1 and type 2 CNV in neovascular AMD cases, except for patients contraindicated by drug hypersensitivity, liver dysfunction, or recent cerebrovascular events. Indocyanine green angiography was also performed along with fluorescein angiography to identify polypoidal choroidal vasculopathy (PCV). In addition, at each patient visit, swept source optical coherence tomography (OCT); Silverstone (NIKON CORPORATION, Tokyo, Japan), DRI OCT Triton (TOPCON CORPORATION, Tokyo, Japan), or spectral domain OCT; NIDEK CO.LTD., Aichi, Japan).

2. Statistical Analysis Statistical analysis was performed using JMP Pro software version 16.2.0 (SAS Institute Inc., Cary, NC). Categorical data were evaluated using Pearson's chi-square test and continuous variables were evaluated using Student's t-test after normalization between endophthalmitis development and control after brolucizumab. After comparing with and without logarithmic transformation, the one better suited to the normal distribution form was used. Because this was a hypothesis-generating study aimed at identifying a possible association between the development of endophthalmitis after brolucizumab and anterior aqueous humor proteins, no Bonferroni correction was performed. Stepwise variable selection (minimum Bayesian information criterion, variable addition) was performed to select explanatory variables for protein concentration. A nominal logistic regression analysis with likelihood ratio test was performed on the incidence of endophthalmitis after brolucizumab. We calculated the area under the receiver operating characteristic (ROC) curve (AUC) and also calculated the sensitivity and specificity of the Youden index.

3. result
Patient demographic characteristics The average age of cases with IOI after brolucizumab (IOI cases), cases without IOI after brolucizumab (non-IOI cases; also referred to as non-IOI cases), and control cases, respectively, was 79.1 years (range, 58-101 years), 75.8 years (57-94 years, P vs. IOI = 0.096), and 75.3 years (60-88 years, P vs AMD = 0.54). 11 of 19 IOI cases (57.9%), 48 of 77 non-IOI cases (62.3%, P for IOI = 0.72), and 14 of 24 control cases (58.3%, P=0.78 vs AMD) were male. The mean duration of treatment for eyes with IOI (IOI eyes) and eyes without IOI (non-IOI eyes) was 33.9 and 43.6 months, respectively (P=0.31). The mean number of injections in IOI and non-IOI eyes was 10.8 and 13.2, respectively (P = 0.43). The mean final injection intervals for IOI, non-IOI, and control eyes were 8 and 10 weeks, respectively (P=0.16). Of the 19 cases of IOI, 13 had PCV, 4 had type 1 CNV, and 2 had RAP, with 5 receiving initial brolucizumab and 14 receiving aflibercept before switching. . No significant difference was seen in the serum 20 vs. 20 cases as shown in the table below.

Protein Concentrations in IOI Patients, Non-IOI Patients, and Control Anterior Aqueous Humor Prior to Brolucizumab Injection Protein concentrations in IOI, non-IOI, and control anterior aqueous humor are shown in FIG. 1B. P-selectin (584 vs 324 pg/mL, P = 0.013), TNF-α (0.89 vs 0.60 pg/mL, P = 0.018), IL-1 α (2.0 vs 1.4 pg/mL, P = 0.035). In addition, IP-10 (61 vs 21 pg/mL, P = 0.15), CXCL13 (22 vs 19 pg/mL, P = 0.27), G-CSF (16 vs 12 pg/mL, P = 0.21), GM- CSF (5.3 vs 4.3 pg/mL, P = 0.49), MMP-9 (127 vs 150 pg/mL, P = 0.46), CXCL1 (48 vs 32 pg/mL, P = 0.16), IL-12 p70 (29 vs 23 pg/mL, P = 0.24), VEGF-A (10 vs 7.4 pg/mL, P = 0.30), IL-17A (1.9 vs 1.6 pg/mL, P = 0.43), IL-1β (1.3 vs 1.0 pg/mL, P = 0.20), E-Selectin (31 vs 25 pg/mL, P = 0.27), ICAM-1 (3536 vs 3155 pg/mL, P = 0.30), IL-2 (6.5 vs 5.7 pg/mL , P=0.17), IFN-γ (2.5 vs 1.8 pg/mL, P=0.086), and MCP-3 (15 vs 21 pg/mL, P=0.37) have P values less than or equal to 0.5.

Protein concentrations in sera of brolucizumab-injected IOI and non-IOI patients. Protein concentrations in IOI and non-IOI sera are shown in FIG. There was a significant difference in the concentration of MMP-9 in IOI patients (16321 vs 10641 pg/mL, P = 0.029) when compared with samples taken from non-IOI patients. In addition, CXCL1 (260 vs 273 pg/mL, P = 0.30), CXCL13 (229 vs 243 pg/mL, P = 0.36), IL-1β (26 vs 29 pg/mL, P = 0.31), IL12 p70 (570 vs 623 pg/mL, P = 0.33) TNF-α (35 vs 37 pg/mL, P = 0.32), G-CSF (260 vs 276 pg/mL, P = 0.41), IL1-α (43 vs 45 pg /mL, P = 0.26), IL-6 (35 vs 38 pg/mL, P = 0.24), IL-17A (15 vs 16 pg/mL, P = 0.46), VEGF-A (59 vs 74 pg/mL , P=0.35) and MCP-3 (63 vs 72 pg/mL, P=0.19) have P values below 0.5.

Fig. 3 shows the percentage of IFNγ and IL4 positive and negative cells and the Th1/Th2 ratio in CD4 positive lymphocyte fraction IOI and non-IOI blood CD4 positive lymphocytes in the blood of IOI patients and non-IOI patients who received brolucizumab injection . show. There was a significant difference in the percentage of Th2 cells among CD4-positive lymphocytes in IOI patients, Th2:IFNγ-/IL4 (3.1 vs 4.2%, P = 0.013) when compared with samples taken from non-IOI patients.

There were significant differences in the concentrations of several cytokines in AMD eyes when compared to controls. MCP-1 (1011 vs 807 pg/mL, P = 0.022*), IP-10 (48 vs 15 pg/mL, P < 0.0001*), CXCL13 (19 vs 13 pg/mL, P = 0.0032*), IL -6 (6.8 vs 19 pg/mL, P = 0.0001*), IL-8 (15 vs 10 pg/mL, P = 0.012*), IL-10 (5.3 vs 2.8 pg/mL, P < 0.0001*), E-selectin (26 vs 4.9 pg/mL, P < 0.0001*), TNF-α (0.65 vs 0.47 pg/mL, P = 0.0033*), and VEGF-A (3.9 vs 25 pg/mL, P < 0.0001*) ). On the other hand, MCP-3 (19 vs 27 pg/mL, P = 0.27), CXCL1 (35 vs 43 pg/mL, P = 0.39), G-CSF (13 vs 14 pg/mL, P = 0.62), GM- CSF (4.5 vs 4.8 pg/mL, P = 0.77), IFN-γ (1.9 vs 1.5 pg/mL, P = 0.19), IL-1 α (1.5 vs 1.7 pg/mL, P = 0.41), IL-1β (1.1 vs 1.1 pg/mL, P = 0.92), IL-5 (0.49 vs 0.36 pg/mL, P = 0.12), IL-2 (5.9 vs 5.6 pg/mL, P = 0.52), IL-12 p70 ( 24 vs 28 pg/mL, P = 0.47), IL-17A (1.6 vs 1.4 pg/mL, P = 0.48), MMP-1 (7.7 vs 5.6 pg/mL, P = 0.090), MMP-9 (145 vs 155 pg/mL, P = 0.72), ICAM-1 (3227 vs 2890 pg/mL, P = 0.25), and P-selectin (364 vs 302 pg/mL, P = 0.37) were not significant.

Nominal logistic regression analysis and ROC curve . , CXCL1 (0.76, 0.071), and IL-5 (0.43, 0.065) were selected by stepwise variable selection. Nominal logistic regression analysis and Youden index yielded "-19 x log10 (MCP-1) + ₁₀ x log10 (IP- ₁₀ ) + 7.2 x _log10 (P-selectin) + 7.4 x _log10 (IL-2 ) + 6.9 x log ₁₀ (TNFα) - 4.3 x log ₁₀ (IL-5) + 16 >0' was found to affect IOI after switching to brolucizumab. AUC was 0.81, sensitivity was 79%, and specificity was 75% (Fig. 4B).

In serum, IL-6 (factor normalized to concentration: -23, P = 0.047), MMP-9 (3.2, 0.036), and IL-1β (19, 0.11) were selected by stepwise variable selection. . By nominal logistic regression analysis and Youden index, “-23 × log ₁₀ (IL-6) + 3.2 × log ₁₀ (MMP-9) + 19 × log ₁₀ (IL-1β)) -8.1 > 0” It turned out to be presumed to affect the IOI after switching. AUC was 0.71, sensitivity was 70%, and specificity was 80% (Fig. 5).

Th2 (P=0.019) was selected by stepwise variable selection in the blood CD4-positive cell fraction. Nominal logistic regression analysis and Youden index found that 'Th2 <3.4' presumed to affect IOI after switching to brolucizumab (where 'Th2 <3.4' refers to the number of Th2 cells in the blood sample). is less than 3.4 percent of the total CD4-positive lymphocyte count.Measurement of the CD4-positive lymphocyte fraction can be performed, for example, by flow cytometry). AUC was 0.71, sensitivity was 73%, and specificity was 77% (Fig. 6A). In addition, the fraction of blood CD4-positive cells and cytokines were Th2 (P = 0.00021), MMP-9 (P = 0.00054), P-selectin (P = 0.0030), CCL7 (P = 0.014), and G-CSF ( P=0.052) were selected by stepwise variable selection. Nominal logistic regression analysis and Youden index indicated that "-1.2 x Th2 + 6.9 x _log10 (MMP-9) + 22 x _log10 (P-selectin) -52 x _log10 (CCL7) -12 x _log10 (G-CSF ) +14 > 0” was found to affect IOI after switching to brolucizumab. AUC was 0.89, sensitivity was 95%, and specificity was 75% (Fig. 6B).

Four. CONCLUSIONS: Patients who develop endophthalmitis after switching to brolucizumab have lower levels of MCP-1 and higher levels of IP-10, CXCL1 and VEGF-A in the anterior aqueous humor before switching. They also have low levels of serum IL-6 and high levels of MMP-9 and IL-1β. In addition, blood Th2 is low. Eyes that develop endophthalmitis after brolucizumab are thought to have asymptomatic cell-mediated immune-mediated inflammation before the switch.

10 Information acquisition part
20 estimation part
30 Notifier
100 estimator
200 system bus
210 CPUs
220 RAM
230 ROMs
240 I/O devices
250 storage devices
260 network connection device

Claims

A method of estimating the likelihood of developing a side effect after administration of brolucizumab in a subject, comprising at least one protein concentration in a body fluid prior to administration of brolucizumab and/or Th2 cell levels in CD4-positive lymphocytes isolated from the subject. A method using a ratio as an index.
The method according to claim 1, wherein the side effect is a side effect associated with intraocular inflammation.
The method according to claim 1 or 2, wherein the side effect is endophthalmitis.
4. The method of any one of claims 1-3, wherein the side effect is selected from the group consisting of uveitis, retinal vasculitis, iritis, inflammation of the anterior chamber, vitreous, and iridocyclitis. .
The method according to any one of claims 1 to 4, wherein the body fluid is selected from the group consisting of blood, plasma, serum, saliva, urine, sweat, tears, vitreous humor, anterior and posterior aqueous humor.
at least one protein wherein the protein is selected from the group consisting of MCP-1, IP-10, P-selectin, CXCL1, VEGF-A, IL-6, MMP-9, IL-1β, CCL7, and G-CSF The method of any one of claims 1-5, comprising
Low MCP-1, high IP-10, high P-selectin, high CXCL1, high VEGF-A, low IL-6, high MMP-9, high IL-1β, low CCL7, low G-CSF , and at least one feature selected from the group consisting of at least one feature selected from the group consisting of a small percentage of Th2 cells in CD4-positive lymphocytes, any one of claims 1 to 6, described method.
8. The method of claims 1-7, wherein the subject has at least one of age-related macular degeneration (AMD), diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity. A method according to any one of paragraphs.
The method of any one of claims 1-8, wherein the subject has been administered aflibercept prior to administration of brolucizumab.
A method of estimating the likelihood of developing endophthalmitis in a subject after administration of brolucizumab, comprising:
Low MCP-1, high IP-10, high P-selectin, high CXCL1, and high VEGF-A in anterior aqueous humor and/or low IL-6, high MMP in serum before brolucizumab administration -9 concentration, and high IL-1β concentration,
of claims 1-9, wherein the subject has at least one of age-related macular degeneration (AMD), diabetic macular edema, retinal vein occlusion, myopic choroidal neovascularization, neovascular glaucoma, and retinopathy of prematurity. A method according to any one of paragraphs.
The formula below:
a×log 10 (MCP-1)＋b×log 10 (IP-10)＋c×log 10 (P-selectin)＋d×log 10 (CXCL1)＋e×log 10 (VEGF-A)＋f＞0
(In the formula, MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A each represent the protein concentration (pg/mL) in body fluids, a = -11 to -0.91, b = 0.50 to 5.2, c = 0.66 to 4.6, d = -0.17 to 4.2, e = 0.0071 to 2.3, and f = -12 to 11)
and/or a × log 10 (IL-6) + b × log 10 (MMP-9) + c × log 10 (IL-1β) + d > 0
(In the formula, IL-6, MMP-9, and IL-1β each represent the serum protein concentration (pg/mL), a = -35 to -12, b = 1.7 to 4.7, c = 7.1 to 30, and d = -17 to 1.2)
and/or blood Th2 < a
(where a = 2.5 to 4.6)
and/or a x Th2 + b x log10 (MMP-9) + c x log10 (P-selectin) + d x log10 (CCL7) + e x log10 (G-CSF) + f > 0
(where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, MMP-9, P-selectin, CCL7, and G-CSF each represent protein concentration in serum (pg/mL), a = -1.6 to -0.78, b = 4.2 to 9.6, c = 12 to 31, d = -76 to -27, e = -19 to -5.3, and f = -12 to 40)
The method according to any one of claims 1 to 10, wherein the possibility of developing a side effect is estimated by
The formula below:
-5.8 x log 10 (MCP-1) + 2.8 x log 10 (IP-10) + 2.6 x log 10 (P-selectin) + 2.0 x log 10 (CXCL1) + 1.1 x log 10 (VEGF- A) - 0.12 > 0
(wherein MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A each represent the protein concentration in the anterior aqueous humor (pg/mL))
and/or −23×log 10 (IL-6) + 3.2×log 10 (MMP-9) +19×log 10 (IL-1β)) −8.1 >0
(wherein IL-6, MMP-9, and IL-1β each represent protein concentration in serum (pg/mL))
and/or blood Th2 < 3.4
and/or -1.2 x Th2 + 6.9 x log10 (MMP-9) + 22 x log10 (P-selectin) -52 x log10 (CCL7) -12 x log10 (G-CSF) +14 > 0
(Where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, and MMP-9, P-selectin, CCL7, and G-CSF are protein concentrations in serum (pg/mL), respectively.)
The method according to any one of claims 1 to 11, wherein the possibility of developing endophthalmitis is estimated by.
A method of estimating the likelihood of developing endophthalmitis in a subject after administration of brolucizumab, comprising:
Concentrations of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A in pre-brolucizumab anterior aqueous humor and/or IL-6, MMP-9, and/or serum isolated from subjects measuring the concentration of IL-1β;
Likelihood of developing endophthalmitis based on measured levels of MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A or levels of IL-6, MMP-9, and IL-1β A method according to any one of claims 1 to 12, comprising the step of estimating .
A kit for use in the method of any one of claims 1-13, comprising:
A kit comprising a capture antibody for an indicator protein immobilized on a solid phase support, and a detection antibody for the indicator protein.
The indicator protein is selected from MCP-1, IP-10, P-selectin, CXCL1, VEGF-A, IL-6, MMP-9, IL-1β, CCL7, and G-CSF, claim 14 Kit as described.
A device for estimating the likelihood of developing a side effect after administration of brolucizumab in a subject, comprising:
an information acquisition unit that acquires information on the percentage of Th2 cells in at least one protein concentration and/or CD4-positive lymphocytes in body fluid before administration of brolucizumab;
An apparatus comprising an estimation unit for estimating the possibility of developing a side effect based on protein concentration and/or information on the ratio of Th2 cells in CD4-positive lymphocytes, and a notification unit for outputting the estimation result.
The estimate is the following formula:
-5.8 x log 10 (MCP-1) + 2.8 x log 10 (IP-10) + 2.6 x log 10 (P-selectin) + 2.0 x log 10 (CXCL1) + 1.1 x log 10 (VEGF- A) - 0.12 > 0
(wherein MCP-1, IP-10, P-selectin, CXCL1, and VEGF-A each represent the protein concentration in the anterior aqueous humor (pg/mL))
and/or −23×log 10 (IL-6) + 3.2×log 10 (MMP-9) +19×log 10 (IL-1β)) −8.1 >0
(wherein IL-6, MMP-9, and IL-1β each represent protein concentration in serum (pg/mL))
and/or blood Th2 < 3.4
and/or -1.2 x Th2 + 6.9 x log10 (MMP-9) + 22 x log10 (P-selectin) -52 x log10 (CCL7) -12 x log10 (G-CSF) +14 > 0
(Where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, and MMP-9, P-selectin, CCL7, and G-CSF are protein concentrations in serum (pg/mL), respectively.)
17. The apparatus of claim 16, performed according to.
A program for estimating the likelihood of developing a side effect after administration of brolucizumab in a subject, comprising:
Obtaining information on the concentration of at least one protein in the body fluid and/or the percentage of Th2 cells in the CD4-positive lymphocytes before administration of brolucizumab;
A step of estimating the possibility of developing side effects based on information on the protein concentration and/or the ratio of Th2 cells in CD4-positive lymphocytes,
A program that executes a process of outputting an estimation result.
The estimate is the following formula:
-5.8 x log 10 (MCP-1) + 2.8 x log 10 (IP-10) + 2.6 x log 10 (P-selectin) + 2.0 x log 10 (CXCL1) + 1.1 x log 10 (VEGF- A) - 0.12 > 0
(wherein MCP-1, IP-10, P-selectin, IL-2, TNFα, and IL-5 each represent the protein concentration in the anterior aqueous humor (pg/mL))
and/or −23×log 10 (IL-6) + 3.2×log 10 (MMP-9) +19×log 10 (IL-1β)) −8.1 >0
(wherein IL-6, MMP-9, and IL-1β each represent protein concentration in serum (pg/mL))
and/or blood Th2 < 3.4
and/or -1.2 x Th2 + 6.9 x log10 (MMP-9) + 22 x log10 (P-selectin) -52 x log10 (CCL7) -12 x log10 (G-CSF) +14 > 0
(Where Th2 is the percentage of Th2 cells in blood CD4-positive lymphocytes, and MMP-9, P-selectin, CCL7, and G-CSF are protein concentrations in serum (pg/mL), respectively.)
19. The program of claim 18, wherein the program is performed based on.
A method of assisting in determining whether or not to administer brolucizumab to a subject, comprising determining the concentration of at least one protein and/or the percentage of Th2 cells among CD4-positive lymphocytes in pre-brolucizumab body fluids isolated from the subject. How to index.
A method of assisting in determining whether concomitant use of an anti-inflammatory agent is necessary when administering brolucizumab to a subject, comprising at least one protein concentration and/or CD4-positive lymphocytes in a body fluid isolated from the subject prior to administration of brolucizumab A method in which the percentage of Th2 cells in the body is used as an index.
A method for estimating the possibility of developing side effects after administration of brolucizumab in a subject, wherein the Th1/Th2 ratio in the blood isolated from the subject before administration of brolucizumab is used as an index.
A computer-readable storage medium that non-transitory stores a program for causing a computer to execute processing including the following steps:
Obtaining information on the concentration of at least one protein in the body fluid and/or the percentage of Th2 cells in the CD4-positive lymphocytes before administration of brolucizumab;
A step of estimating the possibility of developing side effects based on information on the protein concentration and/or the ratio of Th2 cells in CD4-positive lymphocytes,
A step of outputting the estimation result.
A computer-implemented method for estimating the likelihood of developing a side effect in a subject after administration of brolucizumab, comprising:
Obtaining information on the concentration of at least one protein in the body fluid and/or the percentage of Th2 cells in the CD4-positive lymphocytes before administration of brolucizumab;
A step of estimating the possibility of developing side effects based on information on the protein concentration and/or the ratio of Th2 cells in CD4-positive lymphocytes,
A method including the step of outputting an estimation result.
The method according to claim 1, wherein the serum levels of IL-6, MMP-9, and IL-1β are used as indicators.
A method for estimating the possibility of developing side effects after administration of brolucizumab in a subject, using the ratio of Th2 cells among CD4-positive lymphocytes in the blood of the subject as an indicator.
The method according to claim 26, wherein a small proportion of Th2 cells in CD4-positive lymphocytes is used as an index.
The formula below:
Blood Th2 < a (where a = 2.5 to 4.6)
27. The method according to claim 26, wherein the possibility of developing a side effect is estimated by
A method of estimating the likelihood of developing endophthalmitis in a subject after administration of brolucizumab, comprising:
Measuring the concentration of MMP-9, P-selectin, CCL7, and G-CSF in the serum isolated from the subject before administration of brolucizumab, and the percentage of Th2 cells in the blood CD4-positive lymphocytes;
A step of estimating the possibility of developing endophthalmitis based on the measured concentrations of MMP-9, P-selectin, CCL7, and G-CSF, and the percentage of Th2 cells in CD4-positive lymphocytes in the blood The method of any one of claims 1-12, comprising