Method for detecting pulmonary fibrosis

Abstract

The present invention relates to a non-invasive method for detecting pulmonary fibrosis lesions using labeled choline. The present invention also relates to methods for diagnosing the progressive nature, determining the degree of severity and monitoring the progression of progressive pulmonary fibrosis based on this detection method.

Description

[0001] METHOD FOR DETECTING PULMONARY FIBROSIS

[0002] TECHNICAL FIELD OF THE INVENTION

[0003] The present invention relates to a non-invasive method for detecting pulmonary fibrosis lesions. The present invention also relates to methods for diagnosing, determining the degree of severity and monitoring the development of pulmonary fibrosis and its progression.

[0004] TECHNICAL BACKGROUND

[0005] Pulmonary fibrosis (PFF) is a large group of interstitial lung diseases with multiple etiologies. Pulmonary fibrosis can worsen and is then referred to as progressive pulmonary fibrosis (PPF). Collectively, these PFFs have many different etiologies and affect a considerable number of patients (Wijsenbeek et al. Spectrum of Fibrotic Lung Diseases, N Engl J Med. 2020 Sep 3;383(10):958-968).

[0006] Idiopathic pulmonary fibrosis (IPF) is the archetype of PPF and is associated with a progressive decline in lung function with a median survival of less than 5 years after diagnosis (Raghu G et al., Diagnosis of Idiopathic Pulmonary Fibrosis. An Official ATS / ERS / JRS / ALAT Clinical Practice Guideline, Am J Respir Crit Care Med. 2018 Sep I;198(5):e44-e68). IPF is estimated to affect approximately three million patients currently worldwide, with a prevalence between 3 and 45 cases per 100,000 in the general population and an estimated incidence between 0.9 and 13 new cases per 100,000 inhabitants (Martinez FJ et al., Idiopathic pulmonary fibrosis. Nat Rev Dis Primers. 2017 Oct 20;3:17074; Maher et al., Global incidence and prevalence of idiopathic pulmonary fibrosis. Respir Res. 2021;22:197.). Other progressive fibroses are often associated with connective tissue diseases such as rheumatoid arthritis or scleroderma (Wijsenbeek et al., Progressive fibrosing interstitial lung diseases: current practice in diagnosis and management, Curr Med Res Opin. 2019 Nov;35(ll):2015-2024).

[0007] At the mechanistic level, pulmonary fibrosis is the consequence of abnormal scarring with accumulation of extracellular matrix components, particularly collagen fibers, making the lung tissue rigid and non-functional, preventing gas exchange in particular. The exact mechanisms of the initiation and progression of this fibrosis and associated diseases are still poorly understood (Burgy O. et al., Pathogenesis of fibrosis in interstitial lung disease. Curr Opin Pulm Med. 2020 Sep;26(5):429-435).

[0008] Thoracic CT (e.g. high resolution-computed tomography (HR-CT)), a minimally invasive imaging technique, is currently one of the key examinations for diagnosing diffuse interstitial lung disease and for characterizing it, in particular by looking for imaging criteria in favor of fibrosis (honeycomb, traction bronchiectasis in particular).

[0009] However, currently and despite significant advances in scanner techniques, it remains necessary to combine it with an assessment of symptoms and pulmonary function to determine the clinical progression of the disease.

[0010] This assessment of pulmonary function is mainly performed by measuring forced vital capacity (FVC) in order to estimate the severity of the disease. FVC is also one of the main data evaluated during clinical trials (Behr J., Disease Progression in Idiopathic Pulmonary Fibrosis. FVC Is Not Enough, Am J Respir Crit Care Med. 2017 Nov l;196(9):1094-1095). However, the accuracy of FVC measurement is limited and depends on several factors that vary from one patient to another (patient effort, severity of the disease, obstructed airways during measurements).

[0011] In parallel, several potential serum biomarkers of interstitial lung disease and progressive pulmonary fibrosis have been identified (Tzouvelekis A. et al., Validation of the prognostic value of MMP-7 in idiopathic pulmonary fibrosis, Respirology 2017 Apr;22(3):486-493, Ohshimo S. et al., Baseline KL-6 predicts increased risk for acute exacerbation of idiopathic pulmonary fibrosis, Respir Med. 2014 Jul;108(7):1031-9, (Moll et al., Biomark Med. . 2020 Jul;14(ll):997-1007), CXCL13 (Bellamri N. et al., TNF-a and IL-10 Control CXCL13 Expression in Human Macrophages, J Immunol. 2020 May l;204(9):2492-2502, Qiu L. et al., A novel prognostic signature for idiopathic pulmonary fibrosis based on five-immune-related genes, Ann. Transi. Med. 2021 Oct;9(20):1570). Some of these biomarkers are currently being confirmed (e.g. clinical trial NCT04268485 on KL-6) but none are currently usable in routine practice outside of research.

[0012] To date, neither the measurement of forced vital capacity, nor any biomarker, nor imaging allows early and quantitative monitoring of the progression of the disease or the evaluation of the effectiveness of treatments.

[0013] Lung imaging by position emission tomography (PET) with 18 F-fluorodeoxyglucose ( 18 F-FDG) has been studied (T. Win et al., Eur J Nucl Med Mol Imaging. 2012 Mar;39(3):521-8; J. Tanguy et al. Eur J Nucl Med Mol Imaging. 2021 Sep;48(10):3058-3074). However, while it is more expensive and requires a radioactive label, PET imaging 18 F-FDG does not offer any advantage over CT imaging. Therefore, there remains a huge need to develop methods for the diagnosis, prognosis, and monitoring of pulmonary fibrosis such as idiopathic pulmonary fibrosis to accurately assess progression.

[0014] SUMMARY OF THE INVENTION

[0015] The inventors discovered that non-invasive imaging with labeled choline allowed the detection of the progressive nature of pulmonary fibrosis, the assessment of its severity and the prediction of its progression. Indeed, the inventors showed that the signal emitted by the accumulation of labeled choline, a sign of the activity of choline kinase, a key enzyme in choline metabolism and phospholipid production in cells, was directly and positively correlated with the severity of pulmonary fibrosis.

[0016] The present invention therefore relates to a non-invasive method for in vivo detection of pulmonary fibrosis lesion(s) using labeled choline comprising a step of quantifying the signal emitted by the labeled choline in a patient in a determined region, the determined region being all or part of the patient's lungs.

[0017] The present invention also relates to an imaging method comprising a step of quantifying the signal emitted by the labeled choline in a patient in a determined region, the determined region being all or part of the patient's lungs and a step of generating an image from the quantification of the signal emitted by the labeled choline.

[0018] The present invention also relates to the use of labeled choline as an imaging agent in order to detect in vivo pulmonary fibrosis lesions comprising a step of quantifying the signal emitted by the labeled choline in a patient in a determined region, the determined region being all or part of the patient's lungs, and a step of generating an image from the quantification of the signal emitted by the labeled choline.

[0019] The accumulation of labeled choline at the pulmonary level, according to the invention, allows diagnosis, in particular diagnosis of the progressive nature, determination of the degree of severity and / or monitoring of the development of progressive pulmonary fibrosis.

[0020] Also, according to other aspects of the invention, an object of the present invention is labeled choline for use in a method of in vivo diagnosis of progressive pulmonary fibrosis, the method comprising:

[0021] - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs, and

[0022] - a step of comparing the quantity of signal emitted by the labeled choline to a reference value, called the diagnostic reference value, - a quantity of signal emitted by the labeled choline greater than the diagnostic reference value being indicative that the patient is suffering from progressive pulmonary fibrosis.

[0023] Another subject matter of the invention is labeled choline for use in a method for determining in vivo the degree of severity of progressive pulmonary fibrosis, the method comprising:

[0024] - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs, and

[0025] - a step of comparing the quantity of the signal emitted by the labeled choline with at least one reference value, called the grade reference value, each grade reference value corresponding to a given severity grade of progressive pulmonary fibrosis,

[0026] - a quantity of signal emitted by the labeled choline greater than or equal to a grade reference value being indicative that the severity of progressive pulmonary fibrosis is at least of the given severity grade,

[0027] -a quantity of signal emitted by labeled choline lower than the grade reference value being indicative that the degree of severity of progressive pulmonary fibrosis is less than the given severity grade.

[0028] The invention also relates to labeled choline for use in a method of in vivo monitoring of the progression of progressive pulmonary fibrosis, the method comprising:

[0029] (a) a step of quantifying the signal emitted by the labeled choline in a patient in a given region, the given region being all or part of the patient's lungs, at a given time Tl,

[0030] (b) a step of quantifying the signal emitted by the labeled choline in the patient in the region determined at a given time TO, TO being greater than Tl, and

[0031] (c) comparing the amount of signal emitted by choline labeled at T0 to the amount of signal emitted by choline labeled at Tl,

[0032] - a quantity of signal emitted by choline labeled at T0 greater than the quantity of signal emitted by choline labeled at Tl is then indicative of a worsening of progressive pulmonary fibrosis in this patient,

[0033] - a quantity of signal emitted by choline labeled at T0 less than or equal to the quantity of signal emitted by choline labeled at Tl is then indicative of an improvement in progressive pulmonary fibrosis in this patient,

[0034] - a quantity of signal emitted by choline labeled at T0 equal to the quantity of signal emitted by choline labeled at Tl is then indicative of a stagnation of progressive pulmonary fibrosis in this patient.

[0035] According to another aspect, the invention relates to labeled choline for use in a method for in vivo evaluation of the efficacy of a therapeutic treatment against progressive pulmonary fibrosis, the method comprising: (a) a step of quantifying the signal emitted by the labeled choline in a patient in a determined region, the determined region being all or part of the patient's lungs, before therapeutic treatment,

[0036] (b) a step of quantifying the signal emitted by the labeled choline in the patient in the determined region, after a given time of therapeutic treatment and

[0037] (c) comparing the amount of signal emitted by the labeled choline after a given time of therapeutic treatment to the amount of signal emitted by the labeled choline before therapeutic treatment,

[0038] - a decrease or stagnation in the quantity of signal emitted by labeled choline after a given time of therapeutic treatment is indicative of the effectiveness of the therapeutic treatment.

[0039] DETAILED DESCRIPTION OF THE INVENTION

[0040] Method for detecting pulmonary fibrosis lesions

[0041] According to a first aspect, the invention relates to a method for detecting, preferably by imaging, pulmonary fibrosis lesion(s) using labeled choline.

[0042] The invention also relates to labeled choline for use in a method of detecting, preferably by imaging, pulmonary fibrosis lesion(s).

[0043] One or more lesions may be detected.

[0044] Typically, the detection method according to the invention is in vivo. Advantageously, this detection method is non-invasive.

[0045] The invention also relates to the use of labeled choline as an imaging agent in order to detect in vivo one or more pulmonary fibrosis lesions.

[0046] The invention further relates to the use of labeled choline in the preparation of an imaging reagent for the detection of pulmonary fibrosis lesion(s).

[0047] Labeled choline contains a marker. The signal emitted by this marker can be quantified, thus allowing the accumulation of labeled choline in a given tissue, in this case lung tissue, to be quantified. A strong signal from labeled choline indicates a high incorporation of the latter and therefore a high activity of choline kinase and is positively correlated with the fibrosis of said lung tissue.

[0048] Thus, the use of labeled choline according to the invention does not require other molecules than said labeled choline in order to detect pulmonary fibrosis lesions.

[0049] The use of labeled choline as an imaging agent in order to detect in vivo one or more pulmonary fibrosis lesions can therefore be done without another molecule capable of targeting the pulmonary fibrosis lesions. Also, advantageously, the labeled choline is not covalently or non-covalently linked to one or more molecules and / or the labeled choline is not in composition with one or more molecules capable of targeting the pulmonary fibrosis lesions. In particular, according to one embodiment, the labeled choline is not covalently or non-covalently linked and / or in composition with one or more molecules capable of targeting the pulmonary fibrosis lesions, such as for example a peptide binding to CCR2. In all aspects of the present invention, the labeled choline may comprise a label selected from the group consisting of a fluorescent label, a radioactive label and a paramagnetic label.

[0050] The fluorescent marker may be selected from the group consisting of a polymethine dye, such as dicarbocyanine, tricarbocyanine, indotricarbocyanine, merocyanine, styryl, squarilium, holopolar cyanine, hemicyanine, oxonol, and hemioxanol dyes; a rhodamine dye, a phenoxazine dye; or a phenothiazine dye.

[0051] The radioactive marker may be selected from the group consisting of 225 Ac, 211 At, 212 Bi, 213 Bi, 7S Br, n C, 34m CI, 67 Cu, 64 Cu, 18 F, 68 Ga, 166 Ho, 123 L, 124 L, 125 L, 131 L, 13 N, 223 Ra, 186 D, 188 D, 47 Sc, 153 Sm, 94m Tc, " m Tc, and 90 Y, 111 In, 177 Read and 89 Zr.

[0052] The paramagnetic marker can be chosen from the group consisting of Gd3+ , Fe 3+ , Mn 2 * , Yt 3+ , Dy 3+ and Cr 3+ .

[0053] A person skilled in the art can easily determine the most appropriate imaging technique based on the marker chosen.

[0054] Preferably, the labeled choline comprises a radioactive label, more preferably a radioactive label selected from the group consisting of 225 Ac, 211 At, 212 Bi, 213 Bi, 76 Br, U C, 34m CI, 67 Cu, 64 Cu, 18 F, 68 Ga, 166 HO, 123 L, 124 L, 125 L, 131 L, 13 N, 223 Ra, 186 D, 188 D, 47 Sc, 153 Sm, 94m Tc, " m Tc, and 90 Y, m ln, 177 Read, 89 Zr, even more preferably a radioactive label selected from the group consisting of 76 Br, U C, 18 F,123 L, 124 l, and 13 N, most preferably 18 F.

[0055] According to the preferred embodiment of the invention, the labeled choline is a [ 18 F]-Fcholine.

[0056] There [ 18 F]-Fcholine also known as N-[( 18 F)Fluoromethyl]-2-hydroxy-N,N-dimethylethanaminium has the following formula:

[0057] According to the various aspects of the invention, labeled choline, preferably [ 18 F]- Fcholine, is administered to a patient prior to quantification of labeled choline in the patient, preferably 10 to 60 minutes prior to its quantification.

[0058] Administration of labeled choline is usually performed intravenously but can also be performed by any other appropriate method of administration. The dose of labeled choline is determined and adjusted according to factors such as the nature, age, sex and weight of the patient (e.g. dose of 3.7 MBq / kg, 300 MBq maximum).

[0059] The patient may have or be suspected of having progressive pulmonary fibrosis, such as idiopathic pulmonary fibrosis.

[0060] The patient may be a human or non-human animal. Preferably, the patient is a mammal, more preferably the patient is a human being.

[0061] According to usual protocols, when the labeled choline is [ 18 F]-Fcholine and the patient is a human, a dose of 2 to 4 MBq / kg of body mass is administered.

[0062] Advantageously, the detection method according to the invention comprises a step of quantifying the signal emitted by the labeled choline in the patient.

[0063] The signal emitted by labeled choline, more precisely by the label that includes labeled choline, is positively correlated with the metabolic activity of choline kinase in lung tissue. Under the activity of choline kinase, labeled choline is incorporated into the cell membrane. A strong signal of labeled choline reflects a high incorporation of the latter into the cells and therefore a high activity of choline kinase. An increase in the metabolic activity of choline kinase is found in pulmonary fibrosis. Without being bound by theory, the inventors hypothesize that this increase in metabolic activity may be due to an increased synthesis of phosphatidylcholine.

[0064] In all aspects of the present invention, the quantification is preferably carried out in a determined region of the patient. Typically, in the case where pulmonary fibrosis is sought, the determined region is all or part of the patient's lungs.

[0065] The detection method may comprise a step of comparing the quantity of the signal emitted by the labeled choline to at least one reference value, preferably called the detection reference value.

[0066] In one embodiment, the at least one reference value is the quantity of the signal emitted by labeled choline detected by the method according to the invention in a control subject.

[0067] Several reference values ​​can be used in conjunction.

[0068] In one embodiment, the reference value is a so-called basal reference value. The basal reference value is the quantity of the signal emitted by labeled choline detected in a healthy subject (i.e. not presenting pulmonary fibrosis). A quantity of the signal emitted by the labeled choline in the patient greater than said basal reference value is indicative of the presence of pulmonary fibrosis.

[0069] In one embodiment, the at least one reference value is a value obtained in the patient during a previous detection of pulmonary fibrosis using labeled choline.

[0070] In another embodiment, the at least one reference value is the signal emitted by circulating or muscle or liver labeled choline, preferably by circulating labeled choline. The ratio of the signal emitted by the labeled choline relative to that of circulating or muscle or liver choline can then be determined in order to detect pulmonary fibrosis.

[0071] Thus, the accumulation of labeled choline in the lung can also be expressed as the ratio of the pulmonary labeled choline signal to the circulating choline signal. The inventors have shown that the amount of signal emitted by labeled choline is positively correlated with the severity of pulmonary fibrosis.

[0072] In one embodiment, the at least one reference value is a so-called grade reference value. Each given grade reference value corresponds to a given severity grade of pulmonary fibrosis. Each given grade reference value is the amount of signal emitted by labeled choline in a patient having pulmonary fibrosis of that given grade.

[0073] Thus, the detection method according to the invention makes it possible to detect a progressive pulmonary fibrosis lesion but also to determine its severity.

[0074] The detection method according to the invention can be carried out on a patient at different times in time. It thus makes it possible to monitor the development of pulmonary fibrosis in this patient and in particular the severity of the latter. Thus, an increase in the quantity of signal emitted by the labeled choline is indicative of a worsening of the severity of the pulmonary fibrosis. Conversely, a decrease in the quantity of signal emitted by the labeled choline is indicative of a decrease in the severity of the pulmonary fibrosis.

[0075] Monitoring the progression of pulmonary fibrosis can be done in a patient who has been administered therapeutic treatment. The therapeutic treatment may be, for example, pirfenidone or nintedanib or any other therapeutic treatment whose effectiveness on the development of pulmonary fibrosis is to be assessed. A decrease in the amount of signal emitted by labeled choline during the therapeutic treatment is indicative of a patient's response to said therapeutic treatment.

[0076] Advantageously, the detection method may also comprise a step of generating an image from the quantification of the signal emitted by the labeled choline.

[0077] Thus, the generation of an image from the quantification of the signal emitted by choline makes it possible to locate the area(s) of the determined region presenting pulmonary fibrosis.

[0078] Also, the present invention also relates to an imaging method comprising a step of quantifying the signal emitted by the labeled choline in a patient in a determined region, the determined region being all or part of the patient's lungs and a step of generating an image from the quantification of the signal emitted by the labeled choline.

[0079] The step of generating an image from the quantification of the signal emitted by the labeled choline can be done from the quantification of the signal emitted by the labeled choline itself or from the comparison of the quantification of the signal emitted by the labeled choline with a reference value. The imaging equipment used depends on the type of label that the choline comprises. According to the preferred embodiment of the invention where the labeled choline is a [ 18 F]-Fcholine, imaging is performed by positron emission tomography (PET).

[0080] In all aspects of the present invention, imaging of labeled choline may be coupled with other imaging, such as, for example, anatomical imaging such as computed tomography (CT) or magnetic resonance imaging (MRI).

[0081] Method for diagnosing pulmonary fibrosis and in particular its progressive nature

[0082] A second aspect of the invention relates to labeled choline for use in an in vivo diagnostic method of progressive pulmonary fibrosis.

[0083] The invention also relates to a method of diagnosing progressive pulmonary fibrosis in a patient comprising administering labeled choline to said patient.

[0084] The invention also relates to a method for obtaining data to aid in the diagnosis of progressive pulmonary fibrosis comprising a step of imaging labeled choline.

[0085] Furthermore, the invention relates to the use of labeled choline in the preparation of a diagnostic reagent for progressive pulmonary fibrosis.

[0086] Progressive pulmonary fibrosis (PPF) is found in a large number of fibrosing diffuse interstitial lung diseases. Indeed, the most common etiologies of PPF are: idiopathic pulmonary fibrosis, idiopathic non-specific pulmonary fibrosis, pulmonary fibrosis associated with hypersensitivity pneumonitis, iatrogenic pulmonary fibrosis, pulmonary fibrosis secondary to infectious pneumonitis, pulmonary fibrosis secondary to connective tissue diseases such as scleroderma, pulmonary fibrosis associated with rheumatoid arthritis, or pulmonary fibrosis in the context of sarcoidosis.

[0087] PPF may be progressive pulmonary fibrosis associated with rheumatoid arthritis, progressive pulmonary fibrosis associated with systemic sclerosis, progressive pulmonary fibrosis associated with dermatomyositis, or antisynthetase syndrome.

[0088] In all aspects of the present invention, progressive pulmonary fibrosis is selected from the group consisting of idiopathic pulmonary fibrosis (IPF), pulmonary fibrosis secondary to hypersensitivity pneumonitis, iatrogenic pulmonary fibrosis, pulmonary fibrosis secondary to infectious pneumonitis, pulmonary fibrosis secondary to connectives such as systemic sclerosis, pulmonary fibrosis associated with rheumatoid arthritis or pulmonary fibrosis secondary to sarcoidosis.

[0089] According to the preferred embodiment of the invention, the progressive pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0090] According to the preferred embodiment of the invention, the progressive pulmonary fibrosis is a pulmonary fibrosis secondary to a pneumonia of infectious origin. Said pneumonia may be of viral or bacterial origin, preferably viral. Thus, according to one embodiment, the progressive pulmonary fibrosis is a pulmonary fibrosis secondary to a pneumonia selected from the group consisting of influenza-related pneumonia, COVID-19-related pneumonia, pneumococcal infection-related pneumonia and Mycoplasma pneumoniae infection-related pneumonia, preferably pulmonary fibrosis secondary to COVID-19-related pneumonia.

[0091] Typically, the in vivo diagnostic method of progressive pulmonary fibrosis includes the detection of pulmonary fibrosis according to the method described above.

[0092] Advantageously, the method of diagnosis of progressive pulmonary fibrosis includes:

[0093] - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs and

[0094] - a step of comparing the quantity of signal emitted by the labeled choline to a reference value, called the diagnostic reference value.

[0095] Prior to quantifying labeled choline in the patient, labeled choline, preferably [ 18 F]-Fcholine, is administered to a patient.

[0096] The patient is suspected of having progressive pulmonary fibrosis, such as idiopathic pulmonary fibrosis.

[0097] According to one embodiment, an amount of signal emitted by the labeled choline greater than the diagnostic reference value is indicative that the patient has progressive pulmonary fibrosis.

[0098] The diagnostic reference value may be the amount of signal emitted by labeled choline detected in a healthy subject (i.e. not presenting pulmonary fibrosis). An amount of signal emitted by labeled choline in the patient greater than said diagnostic reference value is indicative of the presence of pulmonary fibrosis and therefore that the patient is suffering from progressive pulmonary fibrosis.

[0099] The progressive nature and in particular the imaging evidence of the progression of lung lesions are among the criteria defining progressive pulmonary fibrosis.

[0100] Also, according to another embodiment, the diagnostic reference value corresponds to the quantity of the signal emitted by the labeled choline in the subject suspected of being affected by this pathology at a given time in the past, for example approximately one year before the date of carrying out the diagnostic method according to the invention. A quantity of signal emitted by the labeled choline obtained by the diagnostic method according to the invention greater than the diagnostic reference value is then indicative that the subject is affected by progressive pulmonary fibrosis.

[0101] One aspect of the invention therefore relates to labeled choline for use in a method of in vivo diagnosis of the progressive nature of pulmonary fibrosis.

[0102] The invention also relates to a method for diagnosing the progressive nature of pulmonary fibrosis in a patient comprising administering labeled choline to said patient. The invention also relates to a method for obtaining data to aid in the diagnosis of the progressive nature of pulmonary fibrosis comprising a step of imaging labeled choline.

[0103] The diagnostic method may also comprise a step of generating an image from the quantification of the signal emitted by the labeled choline.

[0104] Thus, generating an image from the quantification of the signal emitted by choline makes it possible to locate the area(s) of the determined region presenting pulmonary fibrosis. Depending on the extent of the area(s) presenting pulmonary fibrosis, the diagnosis of progressive pulmonary fibrosis can be specified.

[0105] According to the preferred embodiment of the invention wherein the labeled choline is a [ 18 F]-Fcholine, the imaging technique used is positron emission tomography (PET).

[0106] Method for determining the severity of progressive pulmonary fibrosis

[0107] A 3rd aspect of the invention relates to labeled choline for use in a method of in vivo determination of the degree of severity of progressive pulmonary fibrosis.

[0108] The invention also relates to a method of determining the degree of severity of progressive pulmonary fibrosis in a patient comprising administering labeled choline to said patient.

[0109] The invention also relates to a method for obtaining data to aid in determining the degree of severity of progressive pulmonary fibrosis comprising a step of imaging labeled choline.

[0110] Furthermore, the invention relates to the use of labeled choline in the preparation of a reagent for determining the degree of severity of progressive pulmonary fibrosis.

[0111] According to the preferred embodiment of the invention, the progressive pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0112] Typically, the method for in vivo determination of the degree of severity of progressive pulmonary fibrosis involves the detection of pulmonary fibrosis according to the method described above.

[0113] Advantageously, the method for determining the degree of severity of progressive pulmonary fibrosis includes:

[0114] - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs, and

[0115] - a step of comparing the quantity of the signal emitted by the labeled choline to at least one reference value, called the grade reference value, each grade reference value corresponding to a given severity grade of progressive pulmonary fibrosis.

[0116] A signal quantity emitted by labeled choline greater than or equal to a grade reference value being indicative that the severity of progressive pulmonary fibrosis is at least of the given severity grade. A signal quantity emitted by labeled choline less than the grade reference value being indicative that the severity of progressive pulmonary fibrosis is less than the given severity grade.

[0117] Prior to quantifying labeled choline in the patient, labeled choline, preferably [ 18 F]-Fcholine, is administered to a patient.

[0118] The patient is a subject with or suspected of having progressive pulmonary fibrosis, such as idiopathic pulmonary fibrosis. Preferably, the patient is a subject with progressive pulmonary fibrosis, such as idiopathic pulmonary fibrosis.

[0119] Each given grade reference value corresponds to a given severity grade of pulmonary fibrosis. Each given grade reference value is the amount of signal emitted by labeled choline in a control patient with progressive pulmonary fibrosis of that given grade.

[0120] The method for determining the degree of severity may also include a step of generating an image from the quantification of the signal emitted by the labeled choline.

[0121] According to the preferred embodiment of the invention wherein the labeled choline is a [ 18F]-Fcholine, the imaging technique used is positron emission tomography (PET).

[0122] Method for monitoring the development of progressive pulmonary fibrosis

[0123] A 4 ème One aspect of the invention relates to labeled choline for use in a method of in vivo monitoring of the progression of progressive pulmonary fibrosis.

[0124] The invention also relates to a method for monitoring the development of progressive pulmonary fibrosis in a patient comprising the administration of labeled choline to said patient.

[0125] The invention also relates to a method for obtaining data to aid in monitoring the progression of progressive pulmonary fibrosis comprising a labeled choline imaging step.

[0126] Furthermore, the invention relates to the use of labeled choline in the preparation of a reagent for monitoring the development of progressive pulmonary fibrosis.

[0127] Typically, the method for monitoring the in vivo evolution of progressive pulmonary fibrosis includes:

[0128] (a) the detection of pulmonary fibrosis according to the method described above in a patient at a given time Tl,

[0129] (b) the detection of pulmonary fibrosis according to the method described above in the patient at a given time, T0 being greater than Tl, and

[0130] (c) comparing the amount of signal emitted by choline labeled at T0 to the amount of signal emitted by choline labeled at Tl.

[0131] Advantageously, the method for monitoring the development of progressive pulmonary fibrosis comprises: (a) a step of quantifying the signal emitted by the labeled choline in a patient in a determined region, the determined region being all or part of the patient's lungs, at a given time Tl,

[0132] (b) a step of quantifying the signal emitted by the labeled choline in the patient in the region determined at a given time TO, TO being greater than Tl, and

[0133] (c) comparing the amount of signal emitted by choline labeled at T0 to the amount of signal emitted by choline labeled at Tl.

[0134] A quantity of signal emitted by choline labeled with TO greater than the quantity of signal emitted by choline labeled with Tl is then indicative of a worsening of progressive pulmonary fibrosis in this patient.

[0135] A quantity of signal emitted by choline labeled at T0 less than or equal to the quantity of signal emitted by choline labeled at Tl is then indicative of an improvement in progressive pulmonary fibrosis in this patient.

[0136] A quantity of signal emitted by choline labeled at T0 equal to the quantity of signal emitted by choline labeled at Tl is then indicative of a stagnation of progressive pulmonary fibrosis in this patient.

[0137] Prior to quantifying labeled choline in the patient, labeled choline, preferably [ 18 F]-Fcholine, is administered to a patient.

[0138] The patient is a subject with or suspected of having progressive pulmonary fibrosis, such as idiopathic pulmonary fibrosis. Preferably, the patient is a subject with progressive pulmonary fibrosis, such as idiopathic pulmonary fibrosis.

[0139] Monitoring the progression of pulmonary fibrosis can be done in a patient who has been administered therapeutic treatment in order to determine the patient's response to the treatment.

[0140] Also, the monitoring method may include a step of administering a therapeutic treatment to the patient. The therapeutic treatment may be, for example, pirfenidone or nintedanib or any other therapeutic treatment whose effectiveness on the development of pulmonary fibrosis is to be assessed.

[0141] A decrease in the amount of signal emitted by labeled choline during therapeutic treatment is indicative of a patient response to said therapeutic treatment.

[0142] The method for monitoring the evolution may also include a step of generating an image from the quantification of the signal emitted by the labeled choline.

[0143] Thus, generating an image from the quantification of the signal emitted by the labeled choline makes it possible to locate the area(s) of the determined region presenting pulmonary fibrosis. Depending on the evolution of the extent of the area(s) presenting pulmonary fibrosis, the evolution of progressive pulmonary fibrosis can be specified.

[0144] According to the preferred embodiment of the invention wherein the labeled choline is a [ 18 F]-Fcholine, the imaging technique used is positron emission tomography (PET). Method for evaluating the effectiveness of a therapeutic treatment against progressive pulmonary fibrosis

[0145] A 5 ème One aspect of the invention relates to labeled choline for use in a method of in vivo evaluation of the efficacy of a therapeutic treatment against progressive pulmonary fibrosis.

[0146] The invention also relates to a method for evaluating the efficacy of a therapeutic treatment against progressive pulmonary fibrosis in a patient comprising the administration of labeled choline to said patient.

[0147] The invention also relates to a method for obtaining data to aid in the evaluation of the efficacy of a therapeutic treatment against progressive pulmonary fibrosis comprising a labeled choline imaging step.

[0148] Furthermore, the invention relates to the use of labeled choline in the preparation of a reagent for evaluating the efficacy of a therapeutic treatment against progressive pulmonary fibrosis.

[0149] Typically, the method for monitoring the in vivo evolution of progressive pulmonary fibrosis includes:

[0150] (a) the detection of pulmonary fibrosis according to the method described above in a patient before therapeutic treatment,

[0151] (b) the detection of pulmonary fibrosis according to the method described above in the patient after a given time of therapeutic treatment, and

[0152] (c) comparing the amount of signal emitted by labeled choline after a given time of therapeutic treatment to the amount of signal emitted by labeled choline before therapeutic treatment.

[0153] Advantageously, the method for in vivo evaluation of the efficacy of a therapeutic treatment against progressive pulmonary fibrosis is characterized in that the method comprises:

[0154] (a) a step of quantifying the signal emitted by the labeled choline in a patient in a determined region, the determined region being all or part of the patient's lungs, before therapeutic treatment,

[0155] (b) a step of quantifying the signal emitted by the labeled choline in the patient in the determined region, after a given time of therapeutic treatment and

[0156] (c) comparing the amount of signal emitted by labeled choline after a given time of therapeutic treatment to the amount of signal emitted by labeled choline before therapeutic treatment.

[0157] A decrease or stagnation in the amount of signal emitted by labeled choline after a given time of therapeutic treatment is indicative of the effectiveness of the therapeutic treatment.

[0158] Prior to quantifying labeled choline in the patient, labeled choline, preferably [ 18 F]-Fcholine, is administered to a patient. The patient is a subject suffering from progressive pulmonary fibrosis, such as idiopathic pulmonary fibrosis.

[0159] Advantageously, the method for evaluating the effectiveness of a therapeutic treatment comprises a step of administering a therapeutic treatment to the patient.

[0160] The amount of signal emitted by labeled choline after a given time of therapeutic treatment can also be compared to the amount of signal emitted by labeled choline after a given time of standard treatment for progressive pulmonary fibrosis such as pirfenidone or nintedanib in a control patient with a pathological profile similar to the patient who received the therapeutic treatment whose efficacy is to be evaluated.

[0161] The method for evaluating the effectiveness of a therapeutic treatment may also include a step of generating an image from the quantification of the signal emitted by the labeled choline.

[0162] According to the preferred embodiment of the invention wherein the labeled choline is a [ 18F]-Fcholine, the imaging technique preferably used is positron emission tomography (PET).

[0163] FIGURES

[0164] Figure 1. Figure 1 shows the quantification via PET imaging at [ 18 F]-Fcholine from lung areas of mice treated with NaCl (white circle) or bleomycin (BLM, black square). The amount of [ 18 F]-Fcholine is expressed as a percentage of the injected dose (ID) by weight (g).

[0165] Statistical analysis was performed using the Mann Whitney test.

[0166] Figure 2. Figure 2 shows the quantification of pulmonary accumulation of [ 18 F]-Fcholine by gamma counting of mice treated with NaCl (white circle) or bleomycin (BLM, black square). Statistical analysis was performed by the Mann Whitney test.

[0167] Figure 3. Figure 3 shows the correlation of the signal [ 18F]-Fcholine at D14 as a function of fibrotic involvement measured by CT scan. Lung density is expressed in Hounsfield units (HU). Statistical analysis was performed by non-parametric correlation with Spearman correlation coefficient calculations *: p<0.05, **: p<0.01.

[0168] Figure 4. Figure 4 shows representative cross-sectional images (n=6 per group) from PET-CT imaging after injection of [ 18 F]-Fcholine to mice treated with bleomycin, with or without nintenanib, or NaCl.

[0169] Figure 5. Figure 5 shows the quantification via PET imaging at [ 18 F]-Fcholine from lung areas of mice treated with NaCl at D0 (white circle), bleomycin at D0 (BLM, black square) or a combination of bleomycin at D0 and nintedanib from D8 to D20 (black triangle). Statistical analysis was performed by the Mann Whitney test.

[0170] Figure 6. Figure 6 shows the correlation of the signal [ 18 F]-Fcholine at D7 as a function of lung involvement between D7 and D21. Lung density is expressed in Hounsfield units (HU). Statistical analysis was performed by non-parametric correlation with Spearman correlation coefficient calculations *: p<0.05, **: p<0.01.

[0171] Figure 7. Figure 7 shows the correlations of the evolution of pulmonary fibrosis (pulmonary delta density) of mice between D7 or D9 and D21 after exposure to bleomycin (BLM, left graphs) or after exposure to bleomycin and treatment with nintedanib (right graphs) with (A) the lung density measured by scanner at D9, (B) the PET signal intensity at [ 18 F]-FDG at D9 or (C) the PET signal intensity at [ 18 F]-Fcholine at D7.

[0172] EXAMPLES induced anger ine

[0173] Materials and methods

[0174] Pulmonary fibrosis was induced in mice by bleomycin administration. Thus, 8-week-old male C57BI / 6 mice were exposed to bleomycin (BLM, n=6) at a dose of 1.25 mg / kg by oropharyngeal aspiration. As a control, 8-week-old male C57BI / 6 mice were treated by intravenous injection of saline (NaCl, n=6).

[0175] The mice were then followed at 7, 14, and 21 days after treatment with bleomycin or NaCl. At each of these times, all mice were injected with [ 18 F]-Fcholine (5 MBq in 100pl per mouse, by intracaudal injection) before undergoing dual positron emission tomography (PET)-thoracic computed tomography (CT) imaging.

[0176] The injection of the [18F ]-Fcholine occurs 10 to 20 minutes before PET imaging. Lung-focused PET imaging lasts 30 minutes (250-700 keV). During the distribution period of the [ 18F]- Fcholine a chest CT scan (CT) is performed (150pA, 45kV, 360 projections, 2 shots / projection). During the procedure, the animals are kept under anesthesia (Isofurane 1.5%) on a heated bed (37°C).

[0177] All PET / CT fusion images were obtained using VivoQuant™ software (Invicro, USA). Each PET / CT image was visually interpreted and 3D regions of interest (3DROI) corresponding to the lungs were semi-automatically drawn for quantification of the tomography and to determine their radioactivity content. Injected doses per animal were measured at the time of injection in MBq. The radioactivity content of the lungs was expressed in MBq, converted into a percentage of the injected dose per gram of tissue (%l D / g). The PET scanner was cross-calibrated with the dose calibrator and the PET data were corrected for scattering and attenuation. All images were corrected for quantification.

[0178] Results

[0179] Signal quantization [ 18 F]-Fcholine showed a significantly higher signal in the lungs of mice with pulmonary fibrosis compared to control mice (Fig. 1).

[0180] The quantification data on imaging was confirmed by gamma counting at D21 (Figure 2).

[0181] The signal difference [ 18 F]-Fcholine between mice with pulmonary fibrosis and control mice is observed for all post-blemoycin time points and the signal accumulates in non-aerated lung areas (fibrous tissue). In addition, the signal [ 18F]-Fchol ine at D14 correlated with the severity of fibrosis measured by chest CT at D21 (Figure 3). This experiment was repeated with an additional group of mice exposed to bleomycin and receiving nintedanib (n=6 mice for each group). This second experiment confirmed the data acquired during the first experiment concerning the difference in signal [ 18 F]-Fcholine between mice treated with saline (NaCl) and bleomycin (BLM) (Figures 4 and 5). Moreover, in animals with pulmonary fibrosis and receiving nintedanib, the signal [ 18 F]-Fcholine is correlated with the severity of fibrotic involvement assessed by chest CT (Figure 6).

[0182] Example 2 - Comparison of CT and PET imaging [ 18 F] FDG and PET at [ 18 Fj-Fchol ine in the evolution of progressive pulmonary fibrosis and its response to treatment

[0183] Materials and methods

[0184] Data A and B from CT and PET imaging at [ 18 F]FDG images shown in Figure 7 were obtained as described in Tanguy et al. Eur J Nucl Med Mol Imaging. 2021 Sep;48(10):3058-3074.

[0185] C data from PET imaging at [ 18 F]-Fcholine shown in Figure 7 were obtained as described in Example 1.

[0186] 8-week-old male C57BI / 6J mice exposed to bleomycin (oropharyngeal aspiration, 1.25 mg / kg) with CT imaging (D7 or D9 or D21), [ 18 F]-FDG (J9) or [ 18 F]-Fcholine (D7). Mice in the “bleomycin + nintedanib” group received nintedanib by oral gavage (60 mg / kg) every day from D9 to D20. Lung density is expressed in Hounsfield units (HU). Statistical analysis was performed by nonparametric correlation with Spearman correlation coefficient calculations. Exact P values ​​are indicated.

[0187] Results

[0188] Our data show that neither the measurement of lung density by CT scan, nor the signal intensity quantified after PET [ 18 F]-FDG, when measured at D9 (beginning of the fibrosis phase) are not predictive of the progression of fibrosis between D9 and D21 post-bleomycin in the experimental model.

[0189] On the contrary, PET imaging at [ 18 F]-Fcholine at D9 is significantly predictive of the evolution of fibrosis observed between D7 and D21 and has a prognostic character of the progression and severity of fibrosis.

[0190] Interestingly, the signal [ 18 F]-Fcholine is also correlated with the anti-fibrotic efficacy of nintedanib in the bleomycin model.

[0191] This shows an interesting superiority of imaging [ 18F]-Fcholine compared to lung density measurements by CT scan (approaching the current standard of care) or PET imaging [ 18 F]-FDG.

Claims

CLAIMS 1. Non-invasive method for in vivo detection of pulmonary fibrosis lesion(s) using labeled choline, characterized in that it comprises: - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs.

2. Imaging method characterized in that it comprises: - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs, and - a step of generating an image from the quantification of the signal emitted by the labeled choline.

3. Use of labeled choline as an imaging agent for detecting pulmonary fibrosis lesions in vivo, characterized in that it comprises: - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs, - a step of generating an image from the quantification of the signal emitted by the labeled choline.

4. Labeled choline for use in an in vivo diagnostic method for progressive pulmonary fibrosis characterized in that the method comprises: - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs and - a step of comparing the quantity of the signal emitted by the labeled choline to a reference value, called the diagnostic reference value, - a quantity of signal emitted by labeled choline greater than the diagnostic reference value being indicative that the patient is suffering from progressive pulmonary fibrosis.

5. Labeled choline for use in a method for in vivo determination of the degree of severity of progressive pulmonary fibrosis characterized in that the method comprises: - a step of quantifying the signal emitted by the labeled choline in a patient in a specific region, the specific region being all or part of the patient's lungs, and - a step of comparing the quantity of the signal emitted by the labeled choline to at least one reference value, called the grade reference value, each grade reference value corresponding to a given severity grade of progressive pulmonary fibrosis - a quantity of signal emitted by the labeled choline greater than or equal to a grade reference value being indicative that the severity of progressive pulmonary fibrosis is at least of the given severity grade, - an amount of signal emitted by labeled choline lower than the grade reference value being indicative that the degree of severity of progressive pulmonary fibrosis is less than the given severity grade.

6. Labeled choline for use in a method for in vivo monitoring of the development of progressive pulmonary fibrosis, characterized in that the method comprises: (a) a step of quantifying the signal emitted by the labeled choline in a patient in a given region, the given region being all or part of the patient's lungs, at a given time Tl, (b) a step of quantifying the signal emitted by the labeled choline in the patient in the region determined at a given time TO, TO being greater than Tl, and (c) comparing the amount of signal emitted by choline labeled at T0 to the amount of signal emitted by choline labeled at Tl, - a quantity of signal emitted by choline labeled at T0 greater than the quantity of signal emitted by choline labeled at Tl is then indicative of a worsening of progressive pulmonary fibrosis in this patient, - a quantity of signal emitted by choline labeled at T0 less than or equal to the quantity of signal emitted by choline labeled at Tl is then indicative of an improvement in progressive pulmonary fibrosis in this patient, - a quantity of signal emitted by choline labeled at T0 equal to the quantity of signal emitted by choline labeled at Tl is then indicative of a stagnation of progressive pulmonary fibrosis in this patient.

7. Labeled choline for use in a method for in vivo evaluation of the efficacy of a therapeutic treatment for progressive pulmonary fibrosis, characterized in that the method comprises: (a) a step of quantifying the signal emitted by the labeled choline in a patient in a determined region, the determined region being all or part of the patient's lungs, before therapeutic treatment, (b) a step of quantifying the signal emitted by the labeled choline in the patient in the determined region, after a given time of therapeutic treatment and (c) comparing the amount of signal emitted by the labeled choline after a given time of therapeutic treatment to the amount of signal emitted by the labeled choline before therapeutic treatment, - a decrease or stagnation in the quantity of signal emitted by labeled choline after a given time of therapeutic treatment is indicative of the effectiveness of the therapeutic treatment.

8. Detection method according to claim 1, imaging method according to claim 2, use of labeled choline according to claim 3 and / or labeled choline for use according to any one of claims 4 to 7 characterized in that the labeled choline comprises a label selected from the group consisting of a fluorescent label, a radioactive label and a paramagnetic label.

9. Detection method according to claim 1, imaging method according to claim 2, use of labeled choline according to claim 3 and / or labeled choline for use according to any one of claims 4 to 7 characterized in that the labeled choline comprises a radioactive marker chosen from the group consisting of 225 Ac, 211 At, 212 Bi, 213 Bi, 76 Br, n C, 34m CI, 67 Cu, 64 Cu, 18 F, 68 Ga, 166 Ho, 123 L, 124 L, 125 L, 131 L, 13 N, 223 Ra, 186 D, 188 D, 47 Sc, 153 Sm, 94m Tc, " m Tc, and 90 Y, 111 1 n, 177 Read and 89 Zr.

10. Detection method according to claim 1, imaging method according to claim 1, use of labeled choline according to claim 3 and / or labeled choline for use according to any one of claims 4 to 7 characterized in that the labeled choline is [ 18 F]-Fcholine.

11. Labeled choline for use according to any one of claims 4 to 1, 8 or 9 characterized in that the progressive pulmonary fibrosis is selected from the group consisting of idiopathic pulmonary fibrosis, idiopathic non-specific pulmonary fibrosis, pulmonary fibrosis associated with hypersensitivity pneumonitis, pulmonary fibrosis of iatrogenic origin, pulmonary fibrosis secondary to pneumonia of infectious origin, pulmonary fibrosis secondary to connective tissue disease, pulmonary fibrosis associated with rheumatoid arthritis, pulmonary fibrosis associated with sarcoidosis, pulmonary fibrosis associated with systemic sclerosis and pulmonary fibrosis associated with dermatomyositis or anti-synthetase syndrome.

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