#1ascendis pharma TransCon™ PTH Top-Line Data from Phase 3 PaTHway Trial March 13, 2022 DRU DRUG#2Cautionary Note on Forward-Looking Statements This presentation contains forward-looking statements. All statements other than statements of historical facts contained in this presentation, such as statements regarding our future results of operations and financial position, including our business strategy, prospective products, clinical trial results, product approvals and regulatory pathways, collaborations, licensing or other arrangements, the scope, support progress, results and costs of developing our product candidates or any other future product candidates, the potential market size and size of the potential patient populations for our product candidates, timing and likelihood of success, plans and objectives of management for future operations, the scope of protection we are able to establish and maintain for intellectual property rights covering our product candidates, and future results of current and anticipated products, are forward-looking statements. These forward-looking statements are based on our current expectations and beliefs, as well as assumptions concerning future events. These statements involve known and unknown risks, uncertainties and other factors that could cause our actual results to differ materially from the results discussed in the forward-looking statements. These risks, uncertainties and other factors are more fully described in our reports filed with or submitted to the Securities and Exchange Commission, including, without limitation, our preliminary prospectus supplement related to the proposed public offering and our most recent Annual Report on Form 20-F filed with the SEC on March 2, 2022 particularly in the sections titled "Risk Factors" and "Management's Discussion and Analysis of Financial Condition and Results of Operations". In light of the significant uncertainties in our forward-looking statements, you should not place undue reliance on these statements or regard these statements as a representation or warranty by us or any other person that we will achieve our objectives and plans in any specified timeframe, or at all. Any forward-looking statement made by us in this presentation speaks only as of the date of this presentation and represents our estimates and assumptions only as of the date of this presentation. Except as required by law, we assume no obligation to update these statements publicly, whether as a result of new information, future events, changed circumstances or otherwise after the date of this presentation. This presentation concerns product candidates that are under clinical investigation and which have not yet been approved for marketing by the U.S. Food and Drug Administration, European Medicines Agency or other foreign regulatory authorities. These product candidates are currently limited by U.S. Federal law to investigational use, and no representations are made as to their safety or effectiveness for the purposes for which they are being investigated. Ascendis, Ascendis Pharma, the Ascendis Pharma logo, the company logo, 2 TransCon, and SKYTROFA are trademarks owned by the Ascendis Pharma group. March 2022 Ascendis Pharma A/S. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#3TransCon PTH PATHway Trial Top-Line Data at Week 26 Pathway TRIAL PaTHway Trial met primary and all key secondary endpoints 78.7% of patients (48 of 61) treated with TransCon PTH achieved the primary endpoint, compared to 4.8% (1 of 21) of patients in the control group (p-value <0.0001) Statistically significant improvements observed on all key prespecified secondary endpoints compared to control: HPES Symptom measures: Physical domain score (p-value = 0.0038) and Cognitive domain score (p-value = 0.0055) HPES Impact measures: Physical Functioning domain score (p-value = 0.0046) and Daily Life domain score (p-value = 0.0061) SF-36v2Ⓡ - Physical Functioning subscale score (p-value = 0.0347) TransCon PTH was generally well tolerated, with no discontinuations related to study drug 82% of TransCon PTH patients and 100% of patients in control group reported treatment-emergent adverse events (TEAEs), the majority of which were Grade 1, 2 in severity. One serious related TEAE in the TransCon PTH arm was reported due to a dosing error One death in the TransCon PTH arm was assessed as unrelated to study drug TransCon PTH-treated patients showed a mean decrease in 24-hour urine calcium excretion into the normal range, from 390 mg/24 hours down to 220 mg/24 hours 3 HPES, Hypoparathyroidism Patient Experience Scales. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#44 Chronic Hypoparathyroidism: Significant Patient Population USA Estimated Prevalence: ~200K in these 3 regions ~70k-112k Europe ~86k-223k Japan ~25k-32k • 2013, Powers et. al., Prevalence and Incidence of Hypoparathyroidism in the United States Using a Large Claims Database, JBMR 2011, Clarke et. al., Co-morbid Medical Conditions Associated with Prevalent Hypoparathyroidism: A Population-Based Study 2013, Underbjerg et. al., Cardiovascular and Renal Complications to Postsurgical Hypoparathyroidism: A Danish Nationwide Controlled Historic Follow- up Study 2015, The Epidemiology of Nonsurgical Hypoparathyroidism in Denmark: A Nationwide Case Finding Study 2016, Astor et. al., Epidemiology and Health-Related Quality of Life in Hypoparathyroidism in Norway 2017. Shishiba et. al., Prevalence of postsurgical hypoparathyroidism in Japan: Estimated from the data of multiple institutes 1999. Nakamura et. al., Prevalence of Idiopathic Hypoparathyroidism and Pseudohypoparathyroidism in Japan Ascendis market research TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#55 Hypoparathyroidism: Multiple Complications Brain fog Laryngospasm & bronchospasm Seizures Anxiety & depression Infections Heart failure Renal failure Numbness & tingling Muscle spasms Fatigue Shobak DM et. al. J Clin Endocrinol Metab. 2016 June 01;101(6):2300-2312. Arrhythmias Abnormal skeletal dynamics Abnormal tissue calcifications Pain TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#6TransCon PTH PATHway (Phase 3) Trial Double-blind, placebo-controlled trial with an open-label extension period adults with chronic hypoparathyroidism randomized 3:1 (TransCon PTH: placebo) Double-Blind Main period (26 weeks) Week 26 PaTHway TRIAL Open-Label Extension period (156 weeks) TransCon PTH 18 mcg/day TransCon PTH (titrated according to algorithm) Placebo TransCon PTH TransCon PTH 6 Placebo Primary Objective Confirm treatment effect of TransCon PTH in adults with hypoparathyroidism Key Eligibility Criteria Adults with chronic hypoparathyroidism (i.e. for at least 26 weeks) Age ≥18 years Reliant on calcitriol ≥0.50 mcg per day or alfacalcidol ≥1.0 mcg per day, and therapeutic elemental calcium ≥800 mg/day for at least 12 weeks prior to screening Serum calcium in normal (or just below normal) range: 7.8-10.6 mg/dL (1.96-2.64 mmol/L) No PTH or PTHrP therapy within 4 weeks prior to Screening Countries Europe (Germany, Denmark, Norway, Italy, Hungary) North America (United States, Canada) 1 No increase in prescribed study drug within 4 weeks prior to Week 26 visit. 2 If needed to meet recommended dietary intake of calcium, it was permitted to take calcium supplements ≤600 mg/day as a nutritional supplement. Primary Composite Endpoint at Week 261 Proportion of patients with: • Serum calcium in the normal range (8.3–10.6 mg/dL) and Independence from active vitamin D and Independence from calcium supplements² Key Secondary Endpoints at Week 26 • HPES Symptom - Physical domain score HPES Symptom - Cognitive domain score • HPES Impact - Physical Functioning domain score HPES Impact - Daily Life domain score SF-36 - Physical Functioning subscale score TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#7PaTHway Trial Patient Disposition Randomized N = 84 Placebo N = 21 ITT POPULATION Placebo Dosed N = 21 TransCon PTH Not Treated N = 2 (1 withdrew consent; 1 recurrence of thyroid cancer) TransCon PTH Discontinued Trial N = 1 (death unrelated to study drug) TransCon PTH N = 63 TransCon PTH Dosed N = 61 • . TransCon PTH Completed Week 26 N = 60 Placebo Completed Week 26 N = 19 PaTHway TRIAL Placebo Discontinued Trial N = 2 (1 withdrew consent; 1 for adverse event of breast cancer) Intention To Treat (ITT): All randomized patients who received at least 1 dose of randomized treatment Safety Analysis Set (SAS): All randomized patients who received at least 1 dose of randomized treatment 7 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#8Patients Who Discontinued Trial during Blinded Treatment Period Randomized Arm Off Study Day Off Study Reason 30 Withdrew consent Placebo Placebo TransCon PTH 62 111 All discontinuations were unrelated to study drug Breast cancer Cardiac arrest TransCon PTH is an investigational product candidate. For investor communication only. 8 Data on file, Ascendis Pharma 2022. Not for use in product promotion. Not for further distribution. ascendis pharma#966 9 ascendis pharma Patient Demographics DRU DRUG#10Demographics and Baseline Characteristics Pathway Characteristics Age (years) (n) TransCon PTH (N = 61) Placebo (N = 21) Mean (SD) 61 49.0 (13.1) 21 47.3 (11.4) Age Group (years) - n (%) <50 28 (45.9) 14 (66.7) ≥50 33 (54.1) 7 (33.3) Sex at Birth n (%) Female 46 (75.4) 18 (85.7) Body Mass Index (kg/m²) (n) Mean (SD) Menopausal Status - n (%) Postmenopausal 61 27.3 (5.8) 46 21 29.5 (5.7) 18 19 (41.3) 3 (16.7) 10 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma TRIAL#11Demographics and Baseline Characteristics (continued) Characteristics Race - n (%) American Indian or Alaska Native Asian Black or African American Native Hawaiian or Other Pacific Islander White Other Geographic Region - n (%) North America Europe Pathway TRIAL TransCon PTH (N = 61) Placebo (N = 21) 0 0 3 (4.9) 2 (9.5) 0 0 57 (93.4) 0 0 19 (90.5) 1 (1.6) 0 39 (63.9) 22 (36.1) 12 (57.1) 9 (42.9) 11 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#12Hypoparathyroidism Disease Etiology and Medical History Characteristics TransCon PTH (N = 61) Placebo (N = 21) Cause of Hypoparathyroidism (HP) Acquired from neck surgery 52 (85.2) 18 (85.7) Autoimmune disease 1 (1.6) 0 Intrinsic genetic defects of the parathyroid glands 3 (4.9) 0 Idiopathic disease 4 (6.6) 3 (14.3) Other 1 (1.6) 0 Duration of HP (Years) (n) 61 21 12.0 11.1 1,56 1,33 Mean Min, Max Patient History Renal Insufficiency History Kidney Stones History Ectopic Calcifications History Vascular Calcifications History Brain Calcification History Cataract History Seizure History 12 Data on file, Ascendis Pharma 2022. 5 (8.2) 1 (4.8) 15 (24.6) 4 (19.0) 0 0 1 (1.6) 1 (1.6) 3 (4.9) 0 0 0 1 (4.8) TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. Pathway TRIAL ascendis pharma#13Baseline Conventional Therapy Conventional Therapy Total Daily Dose (TDD) at Baseline Calcium Supplement/TDD (mg) (n) Mean Min, Max Calcitriol (Active Vitamin D) /TDD (µg) (n) Mean Min, Max Alfacalcidol (Active Vitamin D) /TDD (µg) (n) Mean Min, Max TransCon PTH (N = 61) Placebo (N = 21) 21 61 1748 2105 600, 5000 800, 7200 53 17 0.76 0.69 0.5, 2.0 8 0.5, 1.75 4 2.0 2.5 1.0, 4.0 1.5, 2.5 2 patients did not have eDiary information confirmed by prescription information. 13 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. Pathway TRIAL ascendis pharma#14Baseline Albumin-Adjusted Serum Calcium & 24-Hour Urine Calcium Pathway Lab Summary at Baseline TransCon PTH (N = 61) Placebo (N = 21) 61 21 Albumin-Adjusted sCa (mg/dL) (n) Mean (SD) 24-Hour Urine Calcium (mg/dL) (n) Mean (SD) 8.8 (0.7) 60 8.6 (0.6) 392 (175) 21 329 (140) SCA, serum calcium. 14 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma TRIAL#1515 ascendis pharma Trial Results DRU DRUG#16Primary Composite Endpoint at Week 26 Pathway TRIAL TransCon PTH (N = 61) 48 Placebo (N = 21) 1 4.8% (0.1%, 23.8%) Number of Patients Meeting The Primary Endpoint Criteria at Week 26 (responders) Proportion (95% CI), % Hypothesis Test: p-value (TransCon PTH vs Placebo) 1 Number of Patients Meeting Each Component, (n): Albumin-adjusted sCa within the normal range 2 78.7% (66.3%, 88.1%) Independence from active vitamin D Independence from therapeutic doses of calcium supplements No increase in prescribed study drug Three patients with missing data for at least one of the components are considered as non-responders. <0.0001 49 10 60 5 57 1 57 12 TransCon PTH demonstrated a response rate of 78.7% compared to 4.8% for control (p-value <0.0001) 1 CMH test controlling for etiology of hypoparathyroidism (post-surgical vs other). 2 The normal range for albumin-adjusted sCa is 8.3-10.6 mg/dL (2.07-2.64 mmol/L). Patients with missing data on one or more of the criteria are considered as non- responders. 16 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#17Active Vitamin D Dose (Mean +/- SE) by Visit 1.8 ратнішач TransCon PTH Placebo 1.6 1.4 Active 1.2 Vitamin D 1.0 Dose μg/day Mean (± SE) 0.8 0.6 0.4 0.2 0.0 0 4 8 12 16 20 24 28 Weeks TransCon PTH patients discontinued active vitamin D completely within four weeks 17 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma TRIAL#18Calcium Supplement Dose (Mean +/- SE) by Visit 3000 2500 Calcium Dose 2000 mg/day 1500 Mean (± SE) 1000 500 18 HHIA HH HH HH ратнішач TRIAL TransCon PTH Placebo - 8 12 16 20 24 28 Weeks TransCon PTH enabled rapid and sustained calcium supplement reduction TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#19TRIAL Albumin-adjusted Serum Calcium (Mean +/- SE) by Visit ратнішач 10.0 9.5 Calcium Corrected for 9.0 Albumin mg/dL Mean (± SE) 19 8.5 8.0 7.5 0 HH HH HH H H 8 HH TransCon PTH -Placebo HH 12 16 20 20 Weeks 24 28 ascendis pharma TransCon PTH patients maintained mean serum calcium levels in the normal range at all study visits TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution.#20Key Secondary Endpoints: Patient Reported Symptom & Quality of Life Domains 60 50 40 HPES Symptom Physical domain score 30 20 10 0 HH HH Weeks p-value = 0.0038 50 40 HPES Symptom Cognitive domain score 30 20 10 26 0 Weeks КЯНН p-value = 0.0055 50 26 40 40 HPES Impact Physical Functioning domain score 30 20 20 HH HOH 10 10 0 Weeks p-value = 0.0046 TransCon PTH 50 50 26 HPES Impact Daily Life domain score 40 40 HH 30 30 HH 20 20 10 10 0 Weeks Placebo p-value = 0.0061 55 PaTHway TRIAL SF-36 Physical Functioning subscale score 50 45 45 40 40 35 26 26 Weeks All prespecified key secondary endpoints demonstrated statistically significant improvement compared to control P-values are TransCon PTH vs Control. For HPES, lower scores indicate improvement; for SF-36, higher scores indicate improvement. 20 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. p-value = 0.0347 26 ascendis pharma#21Bone Turnover Markers: P1NP and CTx (Mean +/- SE) by Visit Pathway Mean P1NP TransCon PTH Placebo 150 130 110 Mean CTx TransCon PTH Placebo 1500 Procollagen 1N-Terminal 90 90 Propeptide 70 ng/mL Mean (± SE) 1000 Type I Collagen C- Telopeptides ng/L Mean (± SE) 500 50 30 10 0 4 8 12 16 20 24 28 0 8 12 16 Weeks Weeks Similar pattern exhibited at Week 26 in Phase 2 PaTH Forward Trial P1NP, procollagen type 1 N-terminal propeptide CTX, C-terminal telopeptides of type I collagen 21 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. 20 20 24 24 ascendis pharma TRIAL 28#22ascendis pharma 22 22 Safety Results DRU DRUG#23Overall TEAE Summary ратнішач TRIAL TEAE Summary Treatment-Emergent Adverse Events (TEAE) Serious TEAE TransCon PTH (N = 61); n (%) 50 (82.0) 5 (8.2) Placebo (N = 21); n (%) 21 (100.0) 3 (14.3) Severity* Grade ≥3 2 (3.3) 1 (4.8) Grade 2 21 (34.4) 9 (42.9) 27 (44.3) 11 (52.4) Grade 1 Related TEAE 30 (49.2) 8 (38.1) Serious Related TEAE 1 (1.6) 0 TEAE Related to Hyper- or Hypocalcaemia Leading to ER/Urgent Care Visit and/or Hospitalization 4 (6.6) 2 (9.5) TEAE Leading to Discontinuation of Study Drug 1 (1.6)** 2 (9.5) *In the severity categories, patients are displayed for the highest severity category only. **Death due to cardiac arrest 23 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#24Treatment-Emergent Adverse Events (≥5 patients in total) Preferred Term Patients with at least one TEAE, n (%) TEAEs Injection site reaction Headache Hypocalcaemia Fatigue Paraesthesia Muscle spasms Nausea Arthralgia Diarrhoea Hypercalcaemia Constipation Insomnia 24 Data on file, Ascendis Pharma 2022. Pathway TransCon PTH (N = 61) 50 (82.0) Placebo (N = 21) 21 (100.0) 19 (31.1) 13 (21.3) 0 2 (9.5) 6 (9.8) 9 (42.9) 9 (14.8) 5 (23.8) 11 (18.0) 3 (14.3) 7 (11.5) 3 (14.3) 7 (11.5) 2 (9.5) 6 (9.8) 2 (9.5) 6 (9.8) 1 (4.8) 6 (9.8) 0 4 (6.6) 1 (4.8) 4 (6.6) 1 (4.8) TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma TRIAL#2524-Hour Urine Calcium (mg/d) by Visit p-value <0.0001 Pathway TRIAL 450 TransCon Placebo 400 p-value² =0.24 24-Hour Urine Calcium (mg/d), Change from baseline at Week 26 PTH (N = 61) (N = 21) 350 ANCOVA Model (n) 1 300 24-Hour Urine LS Mean (SE), mg/d 250 Calcium (mg/d) Mean (± SE) 200 95% CI for LS Mean 150 100 Difference in LS Means (SE) 95% CI for Difference in LS Means -154 (21) -64 (32) (-197, -112) (-131,2) -90 (32) (-155,-25) 50 p-value (TransCon PTH vs Placebo) 0.0085 0 Baseline (Week 0) Visit 7 (Week 12) Visit 10 (Week 26) TransCon PTH Placebo 1 The ANCOVA model with unequal variance includes the change from baseline as the response variable, treatment and etiology of HP as fixed effects and baseline value of the parameter as a covariate. 2 p-values from t-test. 25 Data on file, Ascendis Pharma 2022. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#2626 TransCon PTH PATHway Trial Top-line Data at Week 26 ратншач TRIAL PaTHway Trial met primary and all key secondary endpoints 78.7% of patients (48 of 61) treated with TransCon PTH achieved the primary endpoint, compared to 4.8% (1 of 21) of patients in the control group (p-value <0.0001) Statistically significant improvements observed on all key prespecified secondary endpoints compared to control: HPES Symptom measures: Physical domain score (p-value = 0.0038) and Cognitive domain score (p-value = 0.0055) HPES Impact measures: Physical Functioning domain score (p-value = 0.0046) and Daily Life domain score (p-value = 0.0061) SF-36v2 - Physical Functioning subscale score (p-value = 0.0347) TransCon PTH was generally well tolerated, with no discontinuations related to study drug 82% of TransCon PTH patients and 100% of patients in control group reported treatment-emergent adverse events (TEAEs), the majority of which were Grade 1, 2 in severity. One serious related TEAE in the TransCon PTH arm was reported due to a dosing error One death in the TransCon PTH arm was assessed as unrelated to study drug TransCon PTH-treated patients showed a mean decrease in 24-hour urine calcium excretion into the normal range, from 390 mg/24 hours down to 220 mg/24 hours TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ascendis pharma#27Program Status and Next Steps 0 Two Open-Label Extension trials continuing 57 of 59 patients remain in PaTH Forward Trial after two years All 79 patients who completed the blinded period continue in the PaTHway Trial Engage with regulatory authorities regarding registration plans for US and EU Anticipated NDA submission to FDA during Q3, 2022 Anticipated MAA submission to EMA during Q4, 2022 Continue adult TransCon PTH trial in China* Japan Phase 3 top-line data expected in Q3, 2022 Plan to initiate pediatric TransCon PTH trial in Q4, 2022 *In development in Greater China through strategic investment in VISEN 27 Pharmaceuticals. TransCon PTH is an investigational product candidate. For investor communication only. Not for use in product promotion. Not for further distribution. ратнішач TRIAL ascendis pharma#28ascendis pharma Thank you Contact Tim Lee [email protected] DRU DRUG